Abstract:Objective To investigate the effect of miR-214 on the invasive capacity of esophageal squamous carcinoma cell line Eca109, and to explore the possible molecular mechanism. Methods We prepared miR-214 double-stranded mimic and transfected it into Eca109 cells with Lipofectamine 2000. Eca109 cells transfected with nonsense miRNA mimics were taken as control. The expression of mature miR-214 was determined by qPCR. The capacity of cell invasion was determined by Matrigel-coated Transwell assay. E-cadherin protein expression and the percentage of E-cadherin positive cells were examined by Western blotting analysis and flow cytometry, respectively. Results The expression level of mature miR-214 in the miR-214 mimic transfection group was significantly higher than that in the control group 48 h after transfection (P<0.01). Eca109 cells transfected with miR-214 mimic showed a significantly lower cell invasive capacity compared to that of cells transfected with control miRNA mimic (P<0.05). Moreover, E-cadherin protein expression and the ratio of E-cadherin positive cells in miR-214 mimic transfection group were both significantly lower than those in the control group (P<0.05). Conclusion Our data suggest that miR-214 may inhibit the invasive capacity of esophageal squamous carcinoma cells by repressing the epithelial-mesenchymal transition.

Abstract:Objective To construct recombinant adenovirus expressing mouse ChREBP-α, and examine the effect of ChREBP-α overexpression on lipid synthesis in mouse primary hepatocytes. Methods The mouse ChREBP-α cDNA was subcloned into pShuttle-CMV vector, and the product was transformed into E. coli strain BJ5183 for homologous recombination with pAdEasy-1．The resultant recombinant vector was transfected into 293A cells for viral package. Mouse primary hepatocytes were infected with adenoviruses Ad-ChREBP-α, and gene expression was analyzed at mRNA and protein levels by real-time PCR and Western blotting analsysis. The expression levels of ChREBP target gene LPK were measured at mRNA level by real-time PCR, and the fatty acid synthesis rate was determined by \[¹⁴C\]-acetate incorporation. Results We successfully constructed recombinant adenoviruses expressing mouse ChREBP-α. Adenovirus-mediated overexpression of ChREBP-α markedly increased LPK mRNA expression and fatty acid synthesis in primary hepatocytes. Conclusion We have successfully constructed recombinant adenoviruses expressing mouse ChREBP-α, which is biologically active and can overexpress ChREBP-α in primary hepatocytes. Overexpression of ChREBP-α can enhance lipid synthesis.

Abstract:Objective To study the regulatory effect of urocortin (UCN) on ischemia/reperfusion (I/R)-induced myocardial autophagy, so as to explore the myocardial protection mechanism of UCN. Methods Cardiac I/R model was established with rats and hypoxia/reoxygenation(H/R) model was also established with neonatal rat cardiomyocytes. The injury was created by ischemic/hypoxia for 1 h plus reperfusion/reoxygenation for 2 h, and UCN pretreatment was given 1 h before ischemia/hypoxia. The I/R or H/R-induced myocardial injury, myocardial autophagy and autophagy-related gene expression were observed 2 h after reperfusion/reoxygenation. Results UCN pretreatment greatly reduced I/R-induced myocardial damage by decreasing the infarct size, serum creatine kinase (CK) and lactate dehydrogenase (LDH) concentration, increasing the vitality of H/R cardiomyocytes in vitro, and reducing LDH level in the culture supernatant. Moreover, UCN pretreatment also inhibited H/R-induced myocardial autophagy by reducing the ratio of LC3BⅡ/LC3BⅠ and inhibiting expression of autophagy-related genes (Beclin1 and Bnip3). Conclusion UCN can inhibit I/R-induced myocardial autophagy, which may play an important role in the protection against I/R injury.

Abstract:Objective To investigate the origin and protein structures of vertebrate interleukin-17 (IL-17) genes via phylogenetic and bioinformatic approaches. Methods All the known IL-17 protein sequences of human, zebrafish and amphioxus were retrieved from the National Center for Biotechnology Information (NCBI) and Joint Genome Institute (JGI) databases. Also, partial IL-17 sequences from seven other model organisms were collected from several specific databases. MEGA software package was used to reconstruct a phylogenetic tree for these sequences and MODELLER software was used to predict IL-17 three-dimensional models of zebrafish, amphioxus, ciona and caenorhabditis, and these models were then compared with the known crystal structure of human IL-17F. Results Although all the IL-17 protein sequences contained four conservative cysteine residues that may be involved in the formation of a typical cysteine knot, amphioxus IL-17 revealed two different distribution patterns, with one similar to that of vertebrates and one unique to amphioxus. Phylogenetic analysis and cysteine analysis showed that amphioxus genome had no ortholog of vertebrate IL-17 genes, but urochordate ciona genome had it. The structural comparison result of our three-dimensional models supported the phylogenetic tree. Conclusion Amphioxus IL-17 might be the divergent point in the IL-17 evolution. The ortholog of vertebrate IL-17 genes might first appear in urochordates.

Abstract:Objective To compare profiles of microRNA between the LNCaP and LNCaP-AI cell lines, so as to futher elucidate the post-transcritional mechanism regulating the progression to androgen-independence. Methods The microRNA profiles of LNCaP and LNCaP-AI cell lines were examined by Agilent’s microassay. The expression of six microRNAs was verified by RT-PCR. The functions of differentially expressed microRNAs were eludicated by a search with miRBase software (http://www.mirbase.org/). Results The Aglint’s microRNA microassay showed that 11 microRNAs were up-regulated and 27 were down-regulated during the LNCaP progression to androgen-independence. RT-PCR results were consistent with those of the Agilent’s microassay chips. By searching the targets of microRNAs in the miRBase software, we found that the differentically expressed microRNAs were mainly involved in regulation of matrix metalloproteinase 9 (MMP-9), Bcl-2, and epithelial growth factor receptor (EGFR) and genes mediating androgen metabolism. Conclusion There is alteration of microRNA during the progression of LNCaP to androgen-independence, which may involve androgen receptor related pathway, metalloenzyme, anti-apoptotic gene and genes related to androgen metabolism.

Abstract:Objective To evaluate the gastrointestinal (GI) symptoms of peritoneal dialysis (PD) patients and to explore the related factors contributing to GI symptoms. Methods Gastrointestinal Symptom Rating Scale (GSRS) was used for subjective evaluation of GI symptoms in PD patients. The information of patients was obtained by inquiring patients or review of medical records. Multiple regression analysis with stepwise backward variable selection was used to identify factors correlated with GSRS scores. Results The prevalence rate of gastrointestinal symptoms was 61.6% in the PD patients. The prevalence rates of eating dysfunction, reflux and indigestion were 43.8%, 32.1%, and 32.1%, respectively. We found that age (B=0.006, P=0.027), history of corticosteroid therapy (B=0.51，P＜0.001) and daily tablet number (B=0.009, P=0.005) were positively correlated to GSRS score; and residual renal Kt/V (B=-0.27, P=0.001) was negatively correlated with GSRS score. Moreover, females seemed to have more GI symptoms than males (B=-0.15, P=0.033). The above five factors mentioned above could predict 36% of the GSRS score. Conclusion Eating dysfunction, reflux and indigestion are the most common disorders in PD patients. Old age, female sex, more daily tablet number and corticosteroid history are correlated with more severe GI symptoms, while residual renal function might be a preventing factor of GI symptom.

Abstract:Objective To investigate the effect of Shexiangbaoxin pills on the heart function of patients with chronic ischemia heart failure. Methods A total of 96 patients with documented stable chronic ischemia heart failure were evenly randomized into Shexiangbaoxin pill group (Shexiangbaoxin pill plus basic treatment) and placebo group (placebo plus basic treatment) for 24 weeks. The left ventricular ejection fraction (LVEF), left ventricular end diastolic volume (LVEDV), serum brain natriuretic peptide (BNP) level, and 6 minute walk test (6 MWT) were compared before and after treatment in the two groups. Results In Shexiangbaoxin pill group, the LVEF was increased by 5.6% after 24 weeks of treatment (\[34.2±3.5\]% vs \[36.1±4.0\]%, P<0.05), the serum BNP level was significantly decreased (\[680.2±203.9\] pg/mL vs \[621.8±200.7\] pg/mL, P<0.05), and the distance of 6MWT was significantly increased (\[385.3±69.2\] m vs \[401.7±75.0\] m, P<0.05); the LVEDV had no significance before and after treatment (P>0.05). The above parameters were not significantly different in placebo group before and after treatment (P>0.05). Conclusion Shexiangbaoxin pills plus basic treatment can effectively improve the left ventricular function in patients with chronic ischemia heart failure, and further study is needed to observe whether it can reverse left ventricular remodeling.

Abstract:Objective To observe the clinical characteristics of kidney transplant patients combined with coronary heart disease, and to analyze the safety and efficacy of coronary angiography and percutaneous coronary intervention for them. Methods Six kidney transplant patients underwent coronary angiography in our department between 2005 and 2011. The clinical characteristics, coronary angiography data and prognoses of the patients were retrospectively analyzed. Results Three patients had normal coronary angiograms and the other 3 combined with diabetes mellitus also had coronary heart diseases as found by coronary angiography. One case had unstable angina and was implanted with a total of 5 stents during 2 sessions. Another case had acute inferior wall myocardial infarction (caused by complete occlusion of the right coronary artery) was implanted with 2 stents. The other case had acute non-ST-segment elevation myocardial infarction and coronary angiography revealed multi-segment diffuse lesions in 3 arteries; stent implantation failed in this patient. There was no significant difference in renal functions before and after coronary angiography in the 6 patients, and there was no deterioration of renal function during the follow-up. Conclusion Coronary angiography and percutaneous coronary intervention are safe and feasible for kidney transplant patients, and they can greatly improve the quality of life of the patients.

Abstract:Objective To discuss the clinical features, diagnosis and surgical treatment of the primary left ventricular tumors. Methods and results The clinical data of 5 patients with primary left ventricular tumors, who were surgically treated in Changhai Hospital from Jan., 1999 to May,2012, were retrospectively analyzed. The 5 patients accounted for 3.55% (5/141) of patients who were surgically treated for cardiac tumors in the same period. All the 5 cases were diagnosed with echocardiography before treatment. Median sternotomy was performed in 3 cases and left anterolateral thoracotomy in 2 cases. Complete resection of tumor was performed in 3 cases and subtotal resection in 1 case; only biopsy was obtained in the last case. The pathological diagnoses included myxoma, rhabdomyosarcoma, cavernous lymphangioma, papillary elastic fibroma, and intermuscular hemangioma. There was no operative death, and the postoperative course was uneventful. The patients were followed up for 4-80 months (a mean of 30 months) and there was no death. The patient with myxoma had recurrence 38 months after surgery, and there was no recurrence after reoperation. The case with rhabdomyosarcoma survived well; other patients showed no recurrence or metastasis. Conclusion Primary left ventricular tumors are genearally benign. Echocardiography is the most important diagnosis method. The prognosis of malignant left ventricular tumor is poor. The surgical results of left ventricular benign tumors are satisfactory.

Abstract:Objective To study the changes of pleth variability index (PVI) in patients receiving laparoscopic cholecystectomy under general anesthesia, and to investigate the clinical influencing factors of pre-operation baseline PVI value. Methods Totally 67 patients undergoing laparoscopic cholecystectomy with ASA Ⅰ-Ⅱ, aged 18 to 65 years old, were included in the present study. The heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), pulse oximetry (SpO₂), perfusion index (PI), and PVI value were monitored with a Masimo pulse oxygen monitor (Radical-7, USA) after entering the operation room. Fluid expansion with 10 mL/kg of balanced solution was given before the induction. Anesthesia was maintained under total intravenous anesthesia (TIVA) with propofol and remifentanil. Results The average baseline PVI value was (16.8±6.3)% in the patients; then it decreased to (10.3±5.4)% after induction of anesthesia and increased to (21.2±9.5)% after establishment of pneumoperitoneum. The pre-operation baseline PVI value (\[19.2±6.5\] %) of patients <45 years old was significantly higher than that in patients ≥45 years old (\[15.5±5.9\]%, P<0.05%). Male patients had a significantly higher PVI (\[18.8±7.5\]%) value than female patients (\[15.3±4.8\]%, P<0.05). Body mass index (BMI) was not significantly associated with pre-operation baseline PVI. Conclusion The baseline value of PVI is higher than 14%, suggesting insufficient volume load in most of patients. Age and sex are both associated with pre-operation PVI value: male patients and those <45 years old have a higher pre-operation baseline PVI value.

Abstract:Objective To study the role of mestinon-phospholipid complex in promoting the intestinal absorption of mestinon in rats. Methods The mestinon-phospholipid complex was prepared and its composition rate, physico-chemical property and N-octanol/water partition coefficient were examined. Six healthy male rats were randomly divided into two groups. Using the rat model of intestinal absorption, we studied the intestinal absorption rate constant and effective permeability of mestinon-phospholipid complex and free mestinon. Results The composition rate of mestinon-phospholipid complex was (84.02±1.68)%. The characteristic endothermal peak of mestinon disappeared in the DSC curve of mestinon-phospholipid complex. The phase transition temperature of the complex was lower than that of phospholipids, the ultraviolet spectrum profile of the complex was similar to that of mestinon, and the infrared spectrum of the complex was significantly different from that of the physical mixture of mestinon and phospholipid. The particle size of mestinon-phospholipid complex was about (204.6±0.3) nm,and its zeta-potential was (-25.12±0.16) mV. The particle size was largely in a normal distribution. Both the coefficient of recovery and precision of mestinon in intestinal circulating fluid met the requirement. The N-octanol/water partition coefficient of mestinon-phospholipid complex was about 17 folds that of mestinon. The absorption rate constant and the effective permeability of mestinon-phospholipid complex were 2.32 and 2.52 folds those of free mestinon, respectively. Conclusion The mestinon-phospholipid complex can greatly improve the intestinal absorption of mestinon in rats.

Abstract:Objective To assess the association of zinc (Zn) with coal-burning endemic fluorosis (coal-burning fluorosis), so as to provide an evidence for the role of Zn in preventing coal-burning fluorosis. Methods Fifty children with dental fluorosis and 100 healthy controls were selected from Wushan County by stratified sampling, and the same number of children and controls were also selected from Fengjie County. Thirty adults were selected from skeletal fluorosis patients in Duping Township of Wushan County, and 60 healthy controls were selected in the same township. Fasting blood samples were collected from all the participants for Zn determination. Urine samples were collected to examine urine fluoride content. Wilcoxon rank sum test was used to analyze the differences in Zn and fluoride contents between cases and controls and between the two counties. The association between blood Zn and urine fluoride was examined by Pearson linear correlation/Spearman rank correlation. Results The average urine fluoride contents were significantly higher in the child fluorosis cases than in the controls from both counties (P<0.01); and the average blood Zn contents were significantly lower in the fluorosis cases than in the controls from both counties (P<0.01). Moreover, the child cases and controls from Wushan County had significantly lower average blood Zn contents compared with those from Fengjie County (P<0.05); the child fluorosis cases in Wushan County had a significantly lower urine fluoride content compared with those from Fengjie County (P<0.05). The average urine fluoride content of skeletal fluorosis adults from Wushan County was significantly higher than that of controls, and the average blood Zn content was significantly lower than that of controls (P<0.01). In addition, Pearson linear correlation/Spearman rank correlation showed a weak positive relationship between blood Zn and urine fluoride in both children and adults (P<0.05, P<0.01). Conclusion It is indicated that patients with fluorosis are generally Zn deficient, suggesting Zn deficiency might be a risk factor for fluorosis.

Abstract:Objective To study the relationship between adaptability improvement to hot-humid environment with the changes of rectal temperature, heart rate, and scores of comprehensive sensation before and after training in hot-humid environment. Methods Thirty healthy male university students were trained and tested under a temperature of 39.0℃ and a relative humidity of 80.00% for a week. Their rectal temperature, heart rate, and scores of comprehensive sensation were monitored every time before and after training. The obtained data were subjected to ANOVA analysis and Tukey-tests. The improvement rate of rectal temperature and improvement rate of comprehensive sensation scores were calculated each time after training in hot-humid environment. Results and conclusion We found that rectal temperature and scores of comprehensive sensation were consistent in demonstrating the improvement of the adaptability to hot-humid. It is suggested that rectal temperature and scores of comprehensive sensation should be used as reference standards for adaptability improvement to hot-humid environment, and heart rate is not recommended, but the combination of three indices should be used as safety control standard.

Abstract:Objective To screen for the proteins interacting with CXCR4 during nuclear localization in renal cell carcinoma (RCC) A498 cells. Methods Specific band in co-immunoprecipitation (Co-IP) experiments was sent for mass spectrometry. With the results of Co-IP experiments and mass spectrometry, the proteins interacting with CXCR4 were determined by bioinformatics analyses. Results Three specific bands were found after Co-IP with anti-CXCR4 antibody, and the results of mass spectrometry of the three specific bands showed 36 proteins possibly interacting with CXCR4. Bioinformatics analyses showed that NR1D2, c-src and HSPA8 might interact with CXCR4 and participate in CXCR4 nuclear localization. Conclusion NR1D2, c-src and HSPA8 might have participated in CXCR4 nuclear localization in RCC A498 cells.

Abstract:Coagulation disorder, also known as coagulopathy, is commonly associated with traumatic brain injury. The exact mechanisms underlying their association are poorly understood although multiple factors, including abnormalities of coagulation system, hypoperfusion, dilution, metabolic acidosis, hypothermia and inflammation, have been proposed. The abnormalities of coagulation system mainly include tissue factor release, hyperfibrinolysis, and platelet abnormalities. Recently, the dominant role of tissue factor has been challenged by hypoperfusion and the related protein C signaling pathway. The role of inflammation is gradually acknowledged in the late stage of traumatic brain injury-associated coagulopathy. This review was aimed to discuss the above pathogenesis.

Abstract:Colorectal cancer is one of the most common malignant tumors. Previous epidemiological and molecular biological studies have suggested that inflammation plays a pivotal role in the pathogenesis of colorectal cancer. Moreover, many studies have been focused on molecular pathways of inflammation in colorectal cancer. However, the explicit mechanisms involved remain to be fully elucidated. The identification of the key inflammatory mechanisms may provide new targets for intervention and treatment of colorectal cancer. Here we reviewed the inflammatory mechanisms relevant to the carcinogenesis of colorectal cancer, hoping to cast new lights on future research.

Abstract:GCaMPs are new genetically encoded calcium indicators based on a single fluorescent protein molecule with high calcium affinity. The fluorescence intensity of GCaMPs is proportional to intracellular calcium concentration. GCaMPs can be easily targeted and expressed in specific cells and cellular compartments, so as to make it possible for observing calcium-mediated signal transduction. GCaMPs can provide long-term in vivo imaging of calcium signals during development, disease, and drug therapy, making them promising tools for physiological, cytobiological and clinical studies.This paper summarized the principle, progression and application of GCaMPs, hoping to introduce a new calcium detection tool for further exploring the behavior and functions of cells under physiological and pathological states.

Abstract:Objective To introduce the technique of DSA-guided foam sclerotherapy and evaluate its feasibility,safety and curative effects for treatment of lower extremity varicose ulcer (LEVU). Methods A total of 48 legs in 36 selected LEVU patients received DSA-guided foam sclerotherapy with lauromacrogol, with the area of varicose ulcer being 1-7 cm². After high ligation of the great saphenous vein, DSA-guided foam sclerotherapy was performed using 4F catheter to inject the foam of lauromacrogol into the lower extremity varicose to selectively block the great saphenous vein and communicating vein. Ulcer with area larger than 5 cm2 was treated with debridement and skingrafting, and that smaller than 5 cm2 was covered with sterilizing vaseline gauze. All cases wore Ⅰ class compression elastic stockings in the first week after operation and wore Ⅱ class compression elastic stockings from the second week for 3 to 6 months. All patients were given postoperative prophylactic antibiotics and local dressings were changed regularly. Results The procedure was technically successful for all the 48 legs. The injected sclerosant volume ranged from 3.5-8.5 mL for each patient,with an average dose being 6.5 mL; there were no serious complications. The patients could get down the bed and move about on the day after treatment. Two weeks later the ulcer was cured in 38 (79.2%) limbs, reduced in size in 8 (16.7%) limbs, and remained unchanged in 2 (4.2%) limbs. The varicose ulcer disappeared in 45 (93.8%) limbs during 6-12 months follow-up, and 3 (6.2%) limbs had recurrent ulcer. Conclusion DSA-guided foam sclerotherapy of lauromacrogol is a safe, mini-invasive and effective treatment for LEVU, which makes it possible for quick healing of the ulcer after operation.

Abstract:Objective To apply three automatic blood group testing instruments, WADiana^R○ (Diana), Techno TwinStation and ORTHO AutoVue Innova/Ultra, in clinical cross match test and to assess their effect, so as to ensure safe clinical blood transfusion. Methods The three instruments were used for cross matching test of 8 073 blood recipients and blood specimens of Shanghai Blood Center. The incompatible positive samples were reexamined by experiment cross matching antiglobulin test tube method. Results The 3 instruments detected 589 positive blood samples. There was no false positive result in the WADiana^R○(Diana) system; Techno TwinStation and ORTHO AutoVue Innova/Ultra analyzer had 6 and 19 false positive results, respectively. Conclusion All the three instruments can accurately detect the positive samples in clinical cross match test, and WADiana^R○ system has a more prominent performance.

Abstract:Objective To continuously monitor the glucose level during the early phase of acute ischemic stroke (AIS) in patients without diabetes mellitus history, so as to provide evidences for detailed blood glucose information and prognostic evaluation. Methods Ninety-one consecutive non-diabetic AIS patients received continuous glucose monitoring and were divided into normoglycemia (n=20), transient hyperglycemia (n=31), and persistent hyperglycemia (n=40) groups. The general data, blood glucose fluctuation parameters and biochemical indices were compared between the three groups. The National Institutes of Health (NIH) Stroke Scale (NIHSS) scores were also compared between the three groups at 24 h, 7 d, and 14 d after treatment. Results The ratio of male patients and the ratio of cerebral infarction history and hypertension history in the persistent hyperglycemia group were significantly lower than those in the other two groups (P<0.05). Compared with the other two groups, the persistent hyperglycemia group had significantly higher mean blood glucose, standard deviation of blood glucose, largest amplitude of glycemia excursion, mean amplitude of glycemia excursion, and absolute mean of daily differences (all P<0.01). The levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, HbA_1c, C reactive protein, and homocysteine in the persistent hyperglycemia group were significantly higher than those in the other two groups (P<0.01). The NIHSS scores of the persistent hyperglycemia group were significantly higher than those of other groups at 7 d and 14 d after treatment (P<0.05,P<0.01). Conclusion Continuous glucose monitoring can accurately reveal the blood glucose metabolism during the early phase of AIS, contributing the following treatment and prognosis prediction.

Abstract:Objective To compare the PET/CT and CT images of lower extremity long bone osteosarcoma, and to explore their performance in detecting distant/local metastases of osteosarcoma and in clinical staging of the lesions. Methods Fourteen patients with biopsy-confirmed lower extremity long bone osteosarcoma were enrolled in the present study. All patients underwent CT and PET/CT examination before operation. The image characteristic of tumors and bone marrow, lung metastases loci were analyzed. Results The mean maximum standard uptake value (SUV_max) of the primary lesions in this group was 10.68 (ranging 3.7-21.1). PET/CT found that the mean SUV_max value in metastasis group was significantly higher than that in non-metastasis group (11.94 vs 8.01, P<0.05). PET/CT also found 6 cases of bone marrow cavity metastases, and CT only found 2 cases. PET/CT detected 3 cases of lung metastases, and CT only found 2 cases. The accuracy of PET/CT in detecting the bone marrow metastases of osteosarcoma was significantly higher than that of CT (P=0.088 67<0.1). PET/CT showed no significant difference in detecting lung metastases compared with CT. Compared with CT, PET/CT increased the TNM stage of 4 patients (P<0.05). Conclusion Compared with CT, PET/CT has a better performance in demonstrating the tumor involvement, in detecting bone marrow metastasis of osteosarcoma, and in tumor staging. The SUV_max value can help to predict the prognosis of patients with osteosarcoma.

Abstract:Objective To establish a method for simultaneous determination of berberine, magnolol and emodin in the blend powder of Cortex Phellodendri Chinensis, Cortex Magnoliae Officinalis and Radix et Rhizoma Rhei, so as to provide evidence for preparation technique and quality control of simplified recipe of Ruyijinhuangsan. Methods High-performance liquid chromatography (HPLC) was used for simultaneous determination of berberine, magnolol and emodin at the following conditions: the column was ACE.5C₁₈-AR, mobile phase was A: triethylamine, B: methanol. The flow rate was 1.0 mL/min, the temperature of column was 30℃, the detection wavelengths were set at 294 nm and 345 nm, and the injection volume was 10 μL. Using the yield ratio of three constituents as the assessment indices, we optimized the extraction technology by single factor test and orthogonal test L9 (3⁴). Results The calibration curves were linear within the range of 0.292-292 mg/L for berberine (r²=0.999 8), 0.292-292 mg/L for magnolol (r²=0.999 7), and 0.256-256 mg/L for emodin (r²=0.999 5), with the average recovery rates being 98.26% (RSD 2.53%, n=6), 105.17%(RSD 2.06%, n=6), and 100.39% (RSD 2.60%, n=6), respectively. The results of precisions were (n=6) 0.23%, 0.17%, 0.18%, respectively. The sample solutions were stable within 12 h. The single factor experiment analyzed ultrasonic duration and times, concentration of alcohol, the amount of solvent and the concentration of HCl. The results showed that, with the increase of ultrasonic duration, times and the alcohol concentration, the yield ratio of the three consitituents was increased.Conclusion The optimum options for extracting is as follows: ultrasonic extraction 3 times by 95% alcohol containing 0.1% HCl, with the volume at 35 mL, 30 min each time.

Abstract:[ABSTRACT] Objectives To investigate the expression of plasma miR -208b, miR -145 and clinical significance in acute coronary syndrome（ACS）patients. Methods 16 healthy people and 50 ACS patients,including 30 patients with AMI and 20 patients with UAP were enrolled. The plasma samples from ACS patients were obtained within(5±1.4h)（n=50），12h（n=6）and 7d（n=6） after the onset of ACS symtoms. Examing the expression levels of plasma miR -208b and miR -145 by quantitative real-time PCR technology (qRT-PCR) and analyzing the correlation between miR-208b,miR145 expression levels and cTnI,WBC .Results (1) The miR-208b level in plasma from ACS group was higher than that of control group (P <0.05) and the plasma miR208b level in acute myocardial infarction was higher than that of unstable angina group (P <0.05) within (5±1.4h); (2)The miR-145 level in plasma from ACS group was lower than that of in control group(P<0.05) and there is no significant difference of the plasma miR145 expression level in AMI group and UAP group (P >0.05) within (5±1.4h) (3)CTnI which was significantly increased at 6h after ACS remained increased until 12h,and it was restored to the normol value at 7day;both miR208b and miR145 were at their peak values at 6h,and retuned to the control levels at 7d (4)There was positively correlation between of plasma miR208b and cardiac troponin I（cTnI）. (r=0.833, P <0.05)，there was no correlation between plasma miR145 and CTnI（r=0.352，P > 0.05).（5）There was no correlation between of plasma miR208b,145 and WBC（r=-0.243,-0.083，P > 0.05). Conclusion Plasma miR208b is up-regulated and plasma miR145 is down-regulated in ACS. These results suggested that plasma miR208b and miR-145 can be used as a sensitive indicator for ACS.diagnosis.

Abstract:[Abstract] Objective To summarize our experience and operative techniques of transumbilical laparo-endoscopic single site for hydrotubation. Methods Operation was performed on two patients in July and August of 2012．Results The operation was successfully done．The operation time was 55 and 40 min each and the intraoperative blood losses were 100 ml and 50 m1. None of the patients received blood transfusion or antalgic. The drainage was removed 1 days after operation and the postoperative hospital stays were 3 days. Conclusion LESS for infertility is a safe and effective method with less trauma. However, special trainings are needed for the procedure．

Abstract:【Abstract】Obiective:To discuss the the clinical characteristic and the radiological and pathological features of pulmonary lymphomatoid granulomatosis (LYG). Method：We reviewed the clinical materials of the 1 hospitalized patient with LyG diagnosed by pulmonary biopsy through thoracic surgery，analyzed the results of the clinical manifestation，pathology，diagnosis and the therapy． Results：LyG was a kind of the polysystemic disease，which involved in the lung and skin．Chest Radiology was the main method to detect the disease．Pathology was the key method to the diagnosis．Chemotherapy and radiotherapy were effective． Conclusion：LyG is a T-cell rich B-cell lymphoma which is associated with the immunodeficiency and the infection of EB virus；It is rare in the clinic and is very easily misdiagnosed．