Abstract:Objective To examine the feasibility of using photothrombotic unilateral motor cortex lesion for establishing hindlimb spastic hemiplegia model in rats. Methods Twenty SD rats were randomized into 2 groups: rats in group A received erythrosine B injection followed by laser irradiation, and those in group B received sham operation. Then the rate dependent depression (RDD) of H reflex was employed to measure the spasticity of the plantaris before and 3, 7, 14, 28 days post-operation. H-E staining was used to observed the lesions in the motor cortex. Cholera toxin B subunit was used to retrogradely label motoneurons in the spinal cord. The number of vesicular glutamine transporter 1 (VGLUT1) boutons on motoneuron soma and dendrite was quantified by immunofluorescence staining and laser confocal microscope scanning. Results H reflex RDD in the right plantaris muscle was significantly decreased in group A compared with that in group B at 3, 7 ,14, and 28 days post-operation (P < 0.01). However, the H reflex RDD was not significantly different between groups in the left hindlimd at different time points after operation (P > 0.05). The number of VGLUT1 boutons on soma and dendrite in group A was significantly increased compared with that in group B (P < 0.01). H-E staining showed specific lesion in the motor cortex of the brain tissue in group A, but not in group B. Conclusion Unilateral photothrombotic motor cortex lesion is feasible to generate contralateral hindlimb spastic hemiplegia in rats.

Abstract:Objective To explore the factors that are closely related to the sensitivity of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) by analyzing the procedure-related factors, hoping to improve the operation of EBUS-TBNA. Methods A retrospective analysis was performed on all the 393 cases that had received EBUS-TBNA from Jan, 2015 to Dec, 2015 in Changhai Hospital, Second Military Medical University. Positive rates under different conditions such as working experiences of bronchoscopy operators, the types of patients, the types of anesthesia, the positions and sizes of biopsied lymph node, the numbers of biopsied lymph node groups, and the puncture times of each single lymph node were investigated. Logistic regression models and χ²-test were used to identify factors associated with the sensitivity of EBUS-TBNA. Results Our findings showed that the positive rates of EBUS-TBNA were not related to the different working periods as a bronchoscopy operators, types of patients (outpatient or hospitalization), types of anesthesia (local anesthesia or additional intravenous sedation to a local anesthetic), different positions of biopsied lymph node, the puncture times of each single lymph node or the numbers of different groups of lymph node chosen for biopsy. The maximum diameters of punctured lymph nodes under ultrasound interface were found related to the diagnosis rate of EBUS-TBNA (P<0.000 1). Among them, the positive rate of lymph node biopsy was significantly lower in those with the maximum diameter ≤1.0 cm than those with the maximum diameter>2.0 cm (P<0.000 1). The result showed that the positive rate of EBUS-TBNA was positively correlated with the size of biopsied lymph node, with a regression coefficient of 1.027(P<0.001). Conclusion Respiratory physicians can achieve satisfactory performance in EBUS-TBNA operation after formal training and extensive practice. In order to increase the positive rate and to improve the sensitivity of EBUS-TBNA, maximum diameter>2.0 cm lymph nodes should be chosen when possible. The numbers of biopsied lymph node groups, the puncture times of each single lymph node, the positions of biopsied lymph node, the type of anesthesia or the types of patients are not related to the positive rate of EBUS-TBNA.

Abstract:Objective To investigate the correlation of homocysteine (Hcy) level with gene polymorphism of Hcy-metabolizing enzymes in healthy adults receiving physical examination in Shanghai, China. Methods Totally 438 participants who were receiving physical examination in Shanghai Ren-ai Hospital were included in this study. The plasma total Hcy (tHcy) levels and serum folate levels were measured by enzymatic cycling methods and electrochemiluminescence, respectively. Genetic typing was determined by ligase detection reaction(LDR). The plasma tHcy levels were compared between different genotypes, and the association of plasma tHcy levels with genetic polymorphisms was analyzed. Results The incidence of hyperhomocysteinemia (HHcy) was 28.54% (125/438) in the present study. There was significant difference in the levels of plasma tHcy among the three genotypes of MTHFR C677T (P<0.001).Heterozygous genotype CA of MTHFR A1298C was associated with a significantly reduced risk of HHcy (OR=0.49, 95% CI: 0.26-0.92,P=0.027); heterozygous genotype CT of MTHFR C677T was associated with a significantly increased risk of HHcy (OR=2.15 , 95% CI: 1.06-4.36, P=0.035), and homozygous genotype TT was also associated with a significantly increased risk of HHcy (OR=7.58, 95% CI: 3.15-18.22, P<0.001). No correlation was found for HHcy risk with MTR G905A, MTRR A66G, MTRR Ac.56+781C, MTR A2756G, or CBS C551G. Conclusion TT and CT genotype of MTHFR C677T have been found to be the risks for HHcy; CA genotype of MTHFR A1298C may help to decrease the risk to HHcy.

Abstract:Objective To adopt a novel domestic double-disc nitinol occluder to occlude the left atrial appendage of canines and evaluate its feasibility,safety and biocompatibility in experimental canine model. Methods Totally 12 healthy dogs were used in this study; their atrial septal was punctured to occlude the left atial appendage via the femoral vein. The position and effect of the implanted occluders were evaluated by electrocardiogram (ECG) monitoring, atrial septal puncture angiography, transesophageal echocardiography (TEE), and pathological examination immediately, 1, 3, and 6 months after the operation. Results Among the 12 dogs, 11 were successfully implanted with the occluders immediately after operation, with the occluder in 1 case dropped after the operation. Two cases were firstly implanted with occluders of inappropriate size, which were withdrawn and successfully replaced by appropriate ones. The shape and location of the occluders were good as shown by TEE and left atrial angiography examination, with 3 cases having a little shunt and other cases completely occluded immediately after the operation as shown by TEE and left atrial angiography examination. There was no occluder shift, residual shunt or influence on coronary blood flow at 1, 3, and 6 months after the operation. It was shown that the left atrial appendage was completely blocked at 6 months after operation, and there was no thrombus formation on the surface of occluders or in the left atrium; there was no embolization or infarction in the liver,kidney or spleen, with the surface of the occluders completely covered with endothelial cells. Conclusion The novel self-made nitinol occluder has good biological compatibility, making it feasible to occlude the left atrial appendage via transcatheter approach, with satisfactory middle-term effects.

Abstract:Objective To observe the changes of granulocyte-macrophage colony stimulating factor(GM-CSF) expression in mice with radiation and incision combined injury during wound healing and analyze its therapeutic effect, paving a way for exploring the effect of GM-CSF on healing of radiation combination injury. Methods A total of 56 Kunming mice (20-22 g) were randomly divided into two groups: the experimental group and control group (n=28 for each group). Mice in the experimental group were made into irradiated injury plus skin wound by incising the back immediately after exposing to 6 Gy ⁶⁰Co γ radiation. Pure incision injury without radiation exposure was made in the mice in control group. Mice were sacrificed on day 1, 3, 5, and 7 (7 mice for each time) after model establishment, and skin and thin layer of muscle tissues around the wound were collected. Real-time PCR and immunohistochemical analysis were used to examine the mRNA and protein expression of GM-CSF. Moreover, radiation and incision combined injury models were also established in 40 species-, body mass-matched mice using same method described above and the model mice were divided into two groups: treatment group and blank control group (n=20 for each group). The wounds were treated with rhGM-CSF gel (6 000 ng/mL) in the treatment group at 0, 1, 3, 4, 5, 7, 9, 11, and 14 d after model establishment, while blank gel was given to the blank control group. Then the therapeutic effects of rhGM-CSF gel on wound healing were evaluated based on the wound residual area and the level of collagenous fiber around the wound. Results The expression levels of GM-CSF protein and mRNA were significantly lower in the radiation and incision combined injury models during 1-3 days compared with the control group (P<0.05). The expression of GM-CSF was gradually decreased during 5-7 days in the control group, while it kept unchanged in the experimental group. During the 14 days treatment with rhGM-CSF gel, the wound areas were significantly smaller than those of the blank control group (P<0.05). Meanwhile, Masson staining showed notable proliferation, widely distributed, and tightly arranged collagenous fiber after rhGM-CSF gel treatment at the injury sites. Conclusion The level of GM-CSF is lower in radiation and incision combined injury compared with the control, and continuous rhGM-CSF (6 000 ng/mL) treatment can greatly promote the healing of radiation and incision combined injury.

Abstract:Objective To search for the optimal fermentation conditions for producing recombinant Ganoderma lucidum manganese peroxidase (rGlMnP) by Pichia pastoris. Methods Based on the single factorial experiment, an optimum fermentation condition for rGlMnP production by recombinant Pichia pastoris was obtained through a central composite design (CCD) and response surface methodology (RSM) in the present work. The effects of initial pH, temperature, concentrations of methanol and heme on the enzyme activity of rGlMnP produced by Pichia pastoris were explored. The interactive effects and the optimal range levels of the 4 factors were studied and the validity of the model was also verified. Results The results showed that initial pH, temperature, the concentrations of methanol and heme had significant effects on the enzymic activity. The optimum conditions for enzymic activity of rGlMnP produced by Pichia pastoris were as follows: initial pH 5.5,temperature 30℃, concentration of methanol 1.5% and concentration of heme 1.0 mmol/L. Conclusion This study has laid a foundation for producing rGlMnP by Pichia pastoris in industrial scale.

Abstract:Objective To investigate the effect of siomycin A-induced forkhead box protein M1(FoxM1) down-regulation on the malignant behaviors (proliferation, apoptosis, and invasive ability) of laryngeal carcinoma cells. Methods The experiment was performed with Hep-2 cells of the logarithmic growth phase. The experimental groups were treated with different doses of siomycin A, and the control group was treated without siomycin A. CCK-8 test was used to detect the viability of Hep-2 cells after treatment for 24, 48, and 72 h. CFSE staining was used to examine the proliferation ability of Hep-2 cells after treatment for 24 and 48 h. Annexin Ⅴ-FITC/PI dual staining was used to observe the siomycin A-induced apoptosis and Transwell test was used to examine the invasion ability of Hep-2 cells after treatment for 48 h. The expressions of FoxM1, Ki-67 and cleaved caspase-3 protein in Hep-2 cells were detected by Western blotting analysis. The supernatant levels of matrix metalloproteinase (MMP)-2 and MMP-9 were examined by ELISA assay. Results (1) Treatment with siomycin A significantly decreased the expression of FoxM1 protein compared with the control group (P<0.05). CCK-8 test found that siomycin A of different concentrations suppressed the viability of Hep-2 cells in a time- and dose-dependent manner. (2) Siomycin A at 1.3 and 1.5 μmol/L significantly suppressed the proliferation of Hep-2 cells in a dose-dependent manner (P<0.05), accompanied by down-regulated expression of Ki-67.(3)Siomycin A at 1.3 and 1.5 μmol/L also induced greater apoptosis of Hep-2 cells (10.6%, 27.9 %) in a dose-dependent manner compared with control group (4.91%, P<0.05), accompanied by up-regulated cleaved caspase-3 expression. (4) Siomycin A at 1.3 and 1.5 μmol/L suppressed the invasive ability of Hep-2 cells in a dose-dependent manner compared with control group (P<0.05), accompanied by down-regulated expression of MMP-2 and MMP-9. Conclusion Down-regulation of FoxM1 by siomycin A can inhibit the proliferation and invasive ability of laryngeal carcinoma cells.

Abstract:Objective To examine the feasibility of using multiple seasonal autoregressive integrated moving average (SARIMA) model for predicting pulmonary tuberculosis (TB) incidence, so as to provide scientific evidence for the prevention and treatment of TB. Methods EViews 7.0.0.1 software was used to create a SARIMA fit model for seasonal incidence of TB on a monthly basis from January 2004 to December 2012, and the predicting performance of the model was tested with TB data from January to December in 2013. Results The established SARIMA (2,0,2)×(0,1,1)₁₂ model could better fit with the previous TB incidence; and it basically well predicted the TB incidence of the 12 months of 2013, with the mean absolute error being 0.416 992 and the mean absolute error rate being 5.350 8%. Conclusion The established multiplicative SARIMA model can better simulate and predict the trend of TB incidence with time, and it may have a future in predicting the incidence of TB.

Abstract:Objective To investigate the role of Shh-PARP-1 signaling pathway in the protective effects of tea polyphenols against the fatty acid-induced islet microvessel endothelial function injury. Methods Mouse islet microvessel endothelial MS-1 cells were used in this study, and the cells were divided into normal control group, solvent group, fatty acid group (0.25 mmol/L palmitic acid+0.5 mmol/L oleic acid), tea polyphenols group (25 μmol/L), fatty acid+tea polyphenols group, PARP-1 inhibitor (8 μmol/L BYK204165)+fatty acid group, PARP-1 inhibitor+fatty acid+tea polyphenols group, Shh inhibitor (2.5 μmol/L cyclopamine)+fatty acid group, Shh inhibitor+fatty acid+tea polyphenols group and inhibitors of Shh and PARP-1+ fatty acid +tea polyphenols group. The changes of cell viability, apoptosis, nitric oxide (NO) synthesis and oxidative stress related indicators were examined in each group. Results After fatty acid treatment, the survival rate of MS-1 cells was decreased, and the level of apoptosis was significantly increased (P<0.05); meanwhile, fatty acid treatment also significantly increased the content of NO, and the activities of total nitric oxide synthase (tNOS), inducible NOS (iNOS) and constitutive NOS (cNOS) in the cells (P<0.05). Moreover, the lipid peroxidation product, malondialdehyde (MDA) was remarkably elevated and the levels of antioxidants, glutathione (GSH) and superoxide dismutase (SOD), were significantly decreased in response to fatty acid (P<0.05); the expression of PARP-1 and phosphorylated Shh (pShh) was significantly increased (P<0.05). Tea polyphenols could significantly attenuate the toxic effects of fatty acids concerning all the detected indicators (P<0.05). Moreover, after pretreatment with Shh-PARP-1 inhibitors BYK204165 and cyclopamine for 1 h, the protective effects of tea polyphenols were markedly enhanced. There were no significant difference in the detected indicators as compared with controls (P>0.05). Conclusion Fatty acid can directly trigger islet microvessel endothelial function injury, and tea polyphenols shows a protective effect against the toxicity of fatty acid, which can be enhanced by inhibiting Shh-PARP-1 signal pathway.

Abstract:Objective To explore the biocompatibility of rat olfactory ensheathing cells(OECs) with electrospun nanofiber-PLGA (poly[D,L-lactide-co-glycolide]) membrane. Methods The OECs of adult rats were seeded on the electrospun nanofiber-PLGA membrane. The morphology and adhesion to the membrane of OECs were observed by phase contrast microscopy.The proliferation ability of OECs was evaluated by CFDA SE immunofluorescent at 1-5 days after seeding. The control group was seeded into poly-L-lysine-treated plate. Moreover, the electrospun nanofiber-PLGA membrane was also implanted in rats to observe the biocompatibility. Results The purity of OECs obtained from in vitro culture was greater than 90%.The microscopic findings showed that the OECs grew well on the electrospun nanofiber-PLGA membrane, with good adhesion ability and good cell state; the cell also had an orientation growth along the nanofibers.The OECs had a normal proliferation in the electrospun nanofiber-PLGA membrane at 1-5 days after seeding, and the cell number was not significantly different from that of the control group. There was no death in rats after the PLGA film was transplanted, and the PLGA membrane was fused with the surrounding tissues, with some degradation at the fusion sites with the tissues. Conclusion The electrospun nanofiber-PLGA membrane shows a satisfactory biocompatibility, and OECs have good adhesion and proliferation on it, indicating that the membrane might be a potential tissue engineering nerve graft when combined with OECs.

Abstract:Objective To investigate the expression of ubiquitin specific peptidase 7 (USP7) in the sperm of infertile man suffering asthenozoospermia or oligoasthenozoospermia, and to explore its role in the pathogenesis of male infertility. Methods The semen samples were obtained from 120 donors attending the Fertility Clinic, the Centre of Reproductive Medicine, the Second Affiliated Hospital, Chongqing Medical University. The sperm specimens were divided into normozoospermia group (n=57), asthenozoospermia group (n=37) or oligoasthenozoospermia group (n=26) after semen analysis according to 2010 WHO standards. The localization and expression of USP7 were examined using immunofluorescence and Western blotting analysis in the three groups. Then the correlation between USP7 expression and sperm parameters was analyzed. Results USP7 mainly localized in the middle part and main part of the sperm tail; the USP expression was significantly lower in asthenozoospermia group(0.86±0.53) and oligoasthenozoospermia group (0.62±0.43)compared with normozoospermia group (1.63±0.76, P<0.01). And USP7 expression was found positively correlated with the ratio of sperm moving forward(r=0.431 2, P=0.008 7), total motility rate(r=0.443 8, P=0.006 7), and sperm concentration(r=0.455 0, P=0.005 3). Conclusion The lowered expression of USP7 is related to the reduction of human sperm motility and concentration, indicating that USP7 expression may serve as an indicator for sperm quality evaluation in infertile men.

Abstract:Aortic disease(AD) consists of a series of life-threatening diseases caused by aortic wall lesions. Up to now little has been known about the etiology of non-hereditary AD. Vascular smooth muscle cells(VSMC) are the major components of the medial aortic wall and can toggle between a contractile phenotype and a synthetic phenotype. The excessive transformation of VSMC from contractile state to synthetic state plays an important role in the development and progression of AD. Although there are many researches on the regulation of VSMS phenotype switch, how these regulatory pathways operate harmoniously and the what relationship between them remain to be elucidated. Here in this paper we reviewed the existing regulatory mechanisms of VSMC phenotype switch.

Abstract:MicroRNAs (miRs), initially discovered in eukaryotes, are endogenous small noncoding RNAs that regulate gene expression. Myocardial infarction is a common cardiovascular event resulting in cardiac remodeling and subsequent chronic heart failure. A number of miRs have been reported to regulate important pathophysiological processes of myocardial infarction. They have multiple effects on the survival and proliferation of cardiomyocytes, which is of great significance to myocardial infarction and other ischemic heart diseases. In this review we introduced the general knowledge of myocardial infarction and miRs, and described several miRs which are crucial for survival and proliferation of cardiomyocytes. We also discussed the therapeutic potential and limitations of miRs in future treatment of myocardial infarction.

Abstract:Objective To assess the consistencies of VerifyNow system, thrombelastography(TEG), vasodilator-stimulated phosphoprotein (VASP), and PL-11 platelet analyzer in detecting the antiplatelet function of clopidogrel, and to discuss the clinical application values. Methods Totally 98 consecutive inpatients with ST-segment elevation myocardial infarction (STEMI), non-(NSTEMI), or coronary artery in-stent restenosis (ISR) were included in this study. The patients were given a loading dose of 600-mg clopidogrel for 6 hours, 300-mg clopidogrel for at 24 hours, or chronic clopidogrel therapy (75 mg daily for≥7 days) before the procedure. And then the antiplatelet effects were evaluated by VerifyNow, TEG, VASP and PL-11 platelet analyzer simultaneously, with the results of VerifyNow taken as the gold standard and P2Y₁₂ reaction unit (PRU)≥208 taken as high on-clopidogel treatment platelet reactivity (HTPR). Correlation analysis was done for the four methods, and area under ROC curve (AUC) was used to evaluate the value of each method for HTPR. Results The platelet inhibition rate detected by VerifyNow was positively correlated with that by TEG (r=0.234, P<0.05); by contrast, it was negatively correlated with the platelet reactivity index (PRI) measured by VASP, the maximum platelet aggregation rate (MAR) by PL-11 and the maximum adenosine diphosphate (MA-ADP) by TEG (r=-0.299, P<0.01; r=-0.330, P<0.05; r=-0.237, P<0.05). The PRU values was negatively correlated with the platelet inhibition rate detected by VerifyNow (r=-0.815, P<0.01). The area under ROC curve of PL-11 platelet analyzer was the highest (0.644). Conclusion TEG, VASP, and PL-11 platelet function testing systems all have consistency with the "gold standard" VerifyNow in some extent, with PL-11 platelet analyzer showing the highest sensitivity and specificity.

Abstract:Objective To know about the mental health of navy soldiers during long distant sailing and its influencing factors, and to put forward the effective preventive and treatment measures. Methods A questionnaire survey was conducted among 2 987 randomly chosen navy soldiers experiencing long distant sailing. The data collected included general situation, disease prevalence, utilization and cognition of mental health services and mental health state.The structure equation model was used to analyze the influencing factors. Results Totally 2 695 effective questionnaires were collected. Young males were the dominant participants, with a mean age of (25.0±4.3) years old and a mean period of military service of (5.9±4.3) years. It was showed that 76.8% (2 070/2 695) of the soldiers thought that they had no mental illness, 44.5% (1 200/2 695) of the soldiers admitted that they liked army life. The structure equation model analysis showed that the mental health of the navy soldiers was associated with the utilization of mental health service and the physical health was also related to the mental health. Conclusion The navy soldiers experiencing long distant sailing should make full use of the mental health service, and preventive intervention should be given to targeted population so as to improve the mental health of navy soldiers experiencing long distant sailing.

Abstract:Motion sickness is a physiological disorder induced by abnormal acceleration stimuli during spaceflight, aviation or sailing. It has been confirmed that the vestibular system, as the core for locomotion sensing and acceleration information processing, plays key roles in motion sickness initiation and autonomic symptom development. Although the mechanism of motion sickness is still unclear, there have been new breakthroughs in recent years about the physiological basis of sensory conflict theory and the neural mechanism of autonomic reflex. This paper reviewed the progress in the vestibulo-visuo-proprioception information integration, the involvement of vestibulo-hippocampus and vestibulo-cortex in locomotion information processing, vestibulo-autonomic reflex pathway, vestibular neurotransmitter system and anti-motion sickness drug targets, laying a foundation for promoting the development and application of novel countermeasures for motion sickness.

Abstract:Objective To analyze the clinical characteristics, predisposing factors and prognosis of tuberculosis(TB) patients after allogeneic hematopoietic stem cell transplantation(allo-HSCT), and to summarize the clinical treatment experience. Methods The information of 7 confirmed TB patients after allo-HSCT, including the onset time, symptoms,laboratory findings, imaging findings, treatment and prognosis, was collected, and their clinical characteristics and regulations were summarized. Results Seven of the 481 patients who received allo-HSCT were found to have TB, with an post-transplantation TB incidence rate of 1.46%. The seven patients included 2 females and 5 males, ranging 18-46 years and averaging (31.3±9.6) years old. Five patients had pulmonary tuberculosis (2 with hematogenous disseminated pulmonary tuberculosis), 1 had tuberculous pleurisy, 2 had tuberculous meningitis, and 1 had bladder tuberculosis. The clinical manifestations included fever, cough, urinary tract symptom, and headache. Three cases had history of chronic graft versus host disease(GVHD) while one of them experienced acute GVHD. PPD test and tuberculosis antibody test were negative in all the 7 patients: peripheral blood T-SPOT test was positive in 3 cases,with one having positive bronchoalveolar lavage fluid smear. Biopsy results showed granulomatous inflammation in 2 cases, with 1 accompanied by coagulative necrosis. Chest CT showed patched shadow in 3 cases, miliary nodule shadows in 2 and pleural effussion in one. Six patients were clinically cured after anti-tuberculosis treatment and one died, with an effective rate of 85.7%. Conclusion TB infection should be considered once the patients receiving allo-HSCT have infectious symptoms, and early screening should be done to avoid misdiagnosis and missed diagnosis. Early diagnosis and routine anti-TB treatment can yield good outcome.

Abstract:Objective To establish an overload-induced hypertrophy model in aged rats by severing the distal tendon of gastrocnemius muscle, and to investigate the effect of vitamin D on overload-induced hypertrophy and the related mechanism. Methods A total of 20 male rats (24 months old) underwent tenotomy of the achilles tendon of the gastrocnemius muscle in the left hind limb; and a control sham operation was performed on the right hind limb. The rats were randomly divided into control group and experimental group. Experimental group received 1 000 IU/kg of vitamin D by intragastric administration, and the control group was given soybean oil. The animals were sacrificed one week later, the blood samples were collected, and the left, right hind musculus plantaris tissues were weighed and kept in liquid nitrogen. ELISA assay was used to examine serum 25(OH)D level and vitamin D receptor (VDR) in the skeletal muscle. Western blotting analysis was used to examine mTOR, rpS6 protein and their phosphorylation. Results The food intake and body mass were not significantly different between the two groups. Compared with the control side, vitamin D supplement significantly increased the muscle mass of the overload side in both groups (P<0.05); and the mass of the overload side in the vitamin D supplement group was significantly higher than that in the control group (P<0.05), while the mases were not significantly different for the sham sides in the two groups. The results of ELISA assay showed that vitamin D supplement significantly increased serum 25(OH)D levels in rats compared with the control group (P<0.05), and significantly promoted the expression of VDR in the overload side compared with the Sham side (P<0.05) , while there was no significant difference between the two sides in the control group. Western blotting analysis showed that p-rpS6/rpS6 and p-mTOR/mTOR ratios in the overload sides were higher than those in the Sham sides, but significant difference was only found for the vitamin D supplement group (P<0.05). Conclusion Tenotomy of the achilles tendon of the gastrocnemius muscle can effectively promote the skeletal muscle hypertrophy in aging rats, and vitamin D supplement can further enhance overload-induced skeletal muscle hypertrophy, which might be related to VDR expression in skeletal muscle and protein synthesis protein mTOR and rpS6.

Abstract:Objective To explore the protective effect of Jinlida granules on kidney tissues of type 1 diabetic rats, and to elucidate the related mechanism. Methods SD rats were intraperitoneally injected with streptozotocin (STZ, 60 mg/kg) to establish type 1 diabetic models. Then the model rats were randomly divided into diabetic model group, low-, medium- and high-dose Jinlida groups (0.75, 1.5 and 3.0 g/kg Jinlida granules, respectively), Jinlida+Tongxinluo (TXL) group (1.5 g/kg Jinlida granules+0.4 g/kg TXL), metformin group (50 mg/kg metformin), and saxagliptin group (1 mg/kg saxagliptin), with each group containing 5 rats. Five healthy rats served as normal controls. Eight weeks after administration of drugs or placebo, the levels of growth hormone (GH), growth hormone receptor (GHR), insulin-like growth factor 1 (IGF-1), insulin-like growth factor 1 receptor (IGF-1R), and insulin-like growth factor 1 binding protein 1(IGFBP-1) mRNA were detected by real-time quantitative PCR, the expressions of MAPK pathway related proteins and fibronectin (FN) were determined by Western blotting; and H-E staining, Masson staining and PAS staining were used to observe the morphological changes of kidney tissues. Results Compared with the normal control group, the levels of GH, GHR, IGF-1, IGF-1R mRNA and the protein expressions of p-ERK, p-JNK, and FN of kidney tissues were significantly increased (P<0.01), with cellular proliferation and fibrous deposition seen in the kidney tissues in the diabetic model group. After intervention with middle- and high-dose Jinlida granules, the levels of GH, GHR, IGF-1, and IGF-1R mRNA and ratios of p-ERK/ERK, p-JNK/JNK, and FN protein expression were significantly decreased (P<0.05, P<0.01), with alleviated kidney tissues fibrosis. Conclusion Jinlida granules can protect kidney tissues of type 1 diabetic rats, which is probably through up-regulating the levels of GH and IGF-1 mRNA and inhibiting the activation of the MAPK pathway.

Abstract:Objective To investigate the changes of serum progranulin in type 2 diabetic (T2DM) patients with nonalcoholic fatty liver disease (NAFLD) and the relevant clinical significance. Methods The clinical data were subjected to cross-sectional analysis. Totally 116 T2DM patients were randomly selected and assigned to the NAFLD group (n=68) and non-NAFLD group (n=48). The levels of serum progranulin, glycosylated hemoglobin A1c(HbA1c),liver enzymes (ALT, AST, AKP), uric acid (UA), lipids(TG, TC, LDL-C, HDL-C),fasting blood-glucose (FBG), and fasting insulin (FINS) were assessed. Homeostasis model assessment of insulin resistance (HOMA-IR)was calculated. The height, weight, waist circumference and hip circumference were measured, and the body mass index(BMI) and waist-to-hip ratio (WHR) were calculated. The multiple stepwise regression analysis was used to detect the association of progranulin with the above metabolic parameters. The logistic regression was used to analyze the factors influencing NAFLD in patients with T2DM. Results (1)Compared with non-NAFLD group, the levels of BMI, waist circumference, hip circumference, WHR and serum FINS, HOMA-IR, TG, ALT, and AST were significantly higher in the NAFLD group (P<0.05), while the level of HDL-C was significantly lower (P<0.05)in NAFLD group. (2)Serum levels of progranulin were significantly higher in patients with NAFLD than in those with non-NAFLD (P<0.01). (3)Multiple stepwise regression analysis suggested that BMI, HOMA-IR and TG were the independent factors influencing serum progranulin level. (4)Logistic regression analysis showed that progranulin, BMI and ALT were the independent risk factors influencing NAFLD in T2DM patients. Conclusion Serum progranulin level is increased in T2DM patients combined with NAFLD. High level of progranulin is an independent risk factor for the presence of NAFLD in T2DM patients. Progranulin might be an important serological marker of NAFLD in T2DM patients.

Abstract:Objective To examine the expression of cyclophilin A (CypA) in gastric cancer tissues and cell lines, and to explore the effect and mechanism of CypA on the proliferation of gastric cancer cells. Methods Real-time quantitative PCR and Western blotting analysis were used to detect the expression of CypA in gastric cancer tissues, the corresponding adjacent tissues and gastric cancer cell lines. Small interfering RNAs targeting CypA were synthesized and transfected into the gastric cancer cell line MKN45. The effect of CypA-siRNA on endogenous CypA expression was observed; meanwhile, nonspecific siRNA control group and negative control group were also designed. Cell proliferation was measured with MTT assay in each group; cell cycle was detected with flow cytometry. The expression of PCNA, P21, P16, and Cyclin D1 gene, involved in cell proliferation, were also detected by real-time quantitative PCR and Western blotting analysis. Results CypA expression was significantly up-regulated in gastric cancer tissues compared with para-cancer tissues; and CypA expression in gastric cancer cell lines was also significantly higher than that in the gastric epithelial cell line GES-1, with the highest expression found in the poorly-differentiated gastric cancer cell line MKN45(P<0.05). The expression of CypA was significantly inhibited by the CypA-siRNA in MKN45 (P<0.05).MTT assay showed that the inhibition of CypA by CypA-siRNA significantly suppressed MKN45 cell proliferation (P<0.05). The ratio of G₀/G₁ phase cells was significantly increased in MKN45 cells after CypA-siRNA transfection, while the ratio of G₂/M phase was significantly decreased(P<0.05). In addition, the expression of PCNA, Cyclin D1 was significantly decreased at mRNA and protein levels when CypA was inhibited, and the expression of P21 was significantly increased (P<0.05). Conclusion CypA is overexpressed in gastric cancer cells; CypA gene can effectively promote gastric cancer cell proliferation by regulating some proliferation related genes.

Abstract:Objective To investigate the knowledge base and research focuses of stroke rehabilitation via a bibliometrical approach. Methods We searched the Web of Science and identified a total of 5 253 entries on stroke rehabilitation which were published from 2010 to 2015, The literature amount, country distribution, highly cited literature, and high frequency keywords of the 6 years literatures were recorded, and the retrieved literatures were then subjected CiteSpace visualization software analysis to draw the co-citation network clustering map and keyword co-occurrence graph. The basics and research focuses of stroke rehabilitation were analyzed. Results The publications on stroke rehabilitation increased year by year during 2010-2015. The United States, South Korea and Australia were the major countries for stroke rehabilitation research; and the literatures of China had a great increase. The highly cited articles of stroke rehabilitation mainly involved researches about Fugl-Meyer scale,MMSE scale,modified Ashworth scale and Barthel index rating scale and dysfunction recovery. High frequency keyword analysis showed that the researches mainly focused hemiplegia dysfunction in stroke rehabilitation and new techniques and ideas represented by optimized constraint-induced movement therapy and transcranial magnetic stimulation. Conclusion Researches on stroke rehabilitation is in a period of vigorous development, and increasingly more emphasis has been put on stroke rehabilitation research. The highly cited literatures and high frequency keywords can reflect the knowledge base and research focus of stroke rehabilitation, which can be used for further related research.

Abstract:Objective To analyze the characteristics and the prognostic significance of papillary renal cell carcinomas (PRCC). Methods Data about 113 patients with papillary renal cell carcinomas at the time of surgery were retrospectively collected from Changhai Hospital between 2005 and 2015. The prognostic factors were evaluated by Log-rank test．Kaplan-Meier survival curve was used to estimate the survival rate. Results The incidence rate of PRCC was 4.80％ in RCC．And the median age was 54.5 years; M/F ratio was 4.4:1. 55 cases were type I, and 18 were type II, the rest remained unknown.The one-year, 3-year and 5- year survival rates were 100%, 98.5%, 81.3%. The Log rank test showed that type (P=0.035), symptoms at presentation (P=0.008) were associated with survival time. The muhivariable Cox regression model revealed that regional lymph node involvemen（p=0.035）、cancer embolus（p=0.035）were independent predictors of aggressive PRCC. Conlusion PRCC is a distinct type of renal cell carcinoma exhibiting a low degree of malignancy, type II, with initial symptoms indicate worse prognosis.

Abstract:Objective To discuss the curative effect and outcome of the Rh incompatiable allogeneic hematopoietic stem cell transplantation in the treatment of severe aplastic anemia (SAA). Methods One patient with SAA ages 46 (Rh negative) was enrolled in this study in our center on February 7th 2014. The donor is the brother of this patient, with matched HLA and Rh positive blood type. The patient was transplanted with allogeniec hematopoietic stem cell, and the conditioning procedure included fludarabine (FLU), cyclophosphamide (CTX) and rabbit anti-human T-lymphocyte immunoglobulin (ATG). Therapeutic effect of transplantation was reported with its relevant literature reviewed. Results Day 6 after transplantation patient began fever and was diagnosed with klebsiella pneumoniae septicemia. The body temperature returned to normal after therapy. The time of neutrophil and platelet recovery was day 17. The chimeric of bone marrow was 98.3%. Constantly low hemoglobin was treated with infusion and erythropoietin. The red blood cell in bone marrow was proliferated activity on day 31, and the hemoglobin returned to normal gradually. No hemolysis, and the patient received the Rh negative plasma-reduced blood 9 units and platelet 3 units. Up to now we followed this patient for 15 months. The blood and bone marrow were normal, and the chimeric status was 100%, no graft-versus-host disease and other complications occurred. Conclusions Transplantation with Rh incompatiable allogeneic hematopoietic stem cell in treatment of SAA is a safe and effective treatment modality.

Abstract:External root resorption may occur as a consequence of trauma, orthodontic treatment, bacterial infection or incomplete sealing of the root canal system, and lead to crater formation on the resorbed apex. This would deform the root apex surface, and cause loss of apical constriction. Depending on the extent of the resorptive process, different treatment regimens have been proposed. A 31-year-old male patient, right lateral incision gingival fistula, loose 2 degrees, imaging manifestations of external apical root absorption is blunderbuss, the radiograph of apical bone is very discrete, periodontal membrane space widened obviously. In the root canal machinery, chemical cleaning and disinfection of root canal, calcium hydroxide is conveyed to the apical region about 3 mm thick. After the root canal system was filled, the glass ionomer cements and composite resin was filled crownly. These 1-month、3-month、6-month、12-month radiograph showed that the repair of the root absorption area is good, the clinical manifestations of the tooth function is good, no loose. This case shows that calcium hydroxide is successfully used for external apical root resorption cases.