Abstract:Precision medicine has shown a new hope for cancer treatment because it can provide clear individual disease causes, precision therapeutic targets and accurate classification based on the genome sequencing, bioinformatics, and large database. However, the effective precision therapy should also include how to deliver and release drugs with controlled dosage at the precision therapeutic targets. A challenge in clinical anti-tumor precision therapy is how to achieve precision delivery and regulate the dosage of screened drugs, including how to overcome the biological barrier, how to safely deliver the drugs to nidus, how to enhance intratumoral accumulation of drugs, how to promote endocytosis of target cells, and how to accurately deliver drugs to intracellular therapeutic targets. Nanomedicine can effectively solve the problems as mentioned above. Through fine design and nanocarrier tailoring, nano-products will accurately and efficiently deliver the screened drug molecules to target organs, target tissues, target cells or intracellular target organelles, and precisely release the drug to a desirable dosage under the stimulation of lesions. The combination of precision medicine and nanomedicine can efficiently achieve precision therapy: from the tumor molecular classification, drug screening, drug precision delivery, controlled drug release to the treatment. Consequently, precision medicine mainly refers to accurate diagnosis and accurate screening of drug targets, which is the so called "advance troops and scouts" of precision treatment of cancer. Nanomedicine mainly focuss on the accurate delivery and controlled release of drugs to the therapeutic target, which can be called "accurate strategic bombing". Both precision medicine and nanomedicine are essential and interdependent elements in precision therapy.

Abstract:Nanomedicine has shown huge potential in the treatment of cancer, which is not only due to the fact that it can overcome the drawbacks of some small molecular drugs such as insolubility, rapid metabolism, and poor targeting, but also realize the multi modality treatment by containing a variety of molecules. Tremendous novel nano systems referring to above mentioned advantages have been reported so far. However, few newly developed nanomedicine can be approved for clinical applications due to poor stability and rapid metabolism, resulting in the limited efficacy and bottlenecking the development of nanomedicine, that is, "excellent performance in vitro-poor performance in vivo". We believe that the fundamental solution in nanomedicine depends on its basic physical and chemical properties, that is the research and innovation of physical pharmacy theoretical methods of nanopreparation. Herein, focusing on the characteristics of tumor microenvironment and targeting the practical clinical problems, we firstly established a series of physical pharmacy related principles for nanomedicine by the physical and chemical methods combined with the macromolecular Flory-Huggins interaction parameters and scaling theory. Furthermore, taking nanomicelles used to deliver hydrophobic chemotherapeutic drugs as an example, we systemically elaborated the relationship of the physicochemical properties including the carrier composition, self-assembly, drug loading and release, and stability with in vitro and in vivo properties, and explored the related mechanism in vivo.

Abstract:Objective To prepare docetaxel (DTX)-loaded active breast cancer-targeted pH-responsive nanoparticles and to determine its chemo-physical properties, drug loading and releasing characteristics, and targeting ability and cytotoxity against MCF-7 cells. Methods The nanoparticles were synthesized by nanoprecipitation method and surface modification based on polydopamine (PDA). The morphology, size and zeta potential, and surface modification of the nanoparticles were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS) and X-ray photoelectron spectroscopy (XPS), respectively. Drug loading content, encapsulation efficiency, and in vitro drug release profiles were measured by dialysis and high performance liquid chromatography (HPLC). The in vitro cellular uptake was analyzed by confocal laser scanning microscope (CLSM) and flow cytometry (FCM), and the the effect of drug-loaded nanoparticles on the viability of MCF-7 cells was determined by MTT assays. Results The DTX-loaded nanoparticles, CA-PLGA@PDA-PEG-FA/NPs, exhibited a core-shell structure, with hydrodynamic size of (166.4±3.9) nm, zeta potential of (－11.7±3.8) mV, drug loading efficiency of (9.67±0.45)%, and encapsulation efficiency of (88.32±3.10)%. Furthermore, the drug release rate of the nanoparticles in pH 5.0 release medium was faster than that in pH 7.4. XPS spectra showed that PDA and folic acid were modified on the surface of the nanoparticles. The active targeting nanoparticles ingested by MCP-7 cells were more than the nanoparticles not linked to active targeting ligands, and the cytotoxicity of active-targeted nanoparticles was significantly superior than that of Taxotere^® (clinical preparation of DTX). Conclusion The active breast cancer-targeted pH-responsive nanoparticles (DTX-loaded CA-PLGA@PDA-PEG-FA/NPs) exhibits promising targeting ability and efficient antitumor activity in vitro against MCF-7 cells.

Abstract:Objective To prepare a granulocyte targeting-mediated magnetic-fluorescent nanoprobe for detecting prostate cancer PC3 cells in vitro. Methods The nanometer materials with magnetic and red fluorescence, which were prepared using Fe₃O₄ as the core, and SiO₂ and rhodamine isothiocyanate as the shell, were mixed with normal human peripheral blood granulocytes in different proportions, and co-incubated for different periods to examine the toxicity of nanometer materials to granulocytes. The best proportion was selected to combine the nanometer materials and granulocytes in vitro. Finally we obtained the granulocyte targeting-mediated magnetic-fluorescent nanoprobes. We mixed PC3 cells and normal human whole blood cells in different proportions, added the nanoprobes, and then observed the targeting situation of the nanoprobes under a fluorescence microscope. Results The nanoprobe had no obvious influence on the survival rate of granulocytes at different concentrations and action times set in this study. The nanoprobes were enriched around the PC3 cells with a "petal-like" structure, but the peripheral blood cells were not enriched by probes. Conclusion The magnetic-fluorescent nanometer materials prepared in this study have no toxicity to granulocytes, and it can effectively detect tumor cells by the biological targeting effect of granulocytes on tumor cells.

Abstract:Objective To prepare antibody-targeting temperature-sensitive polylated N-isopropylacrylamide/polyethyleneimine (PNIPAM/PEI) nanogel for delivering siRNA against ribonucleotide reductase (RR) subunit M2 (RRM2, RRM2-siRNA) and to establish a new targeted nano-gel delivery system for anti-tumor therapy by studying its anti-tumor ability in vitro. Methods We synthesized the core-shell temperature-sensitive PNIPAM/PEI nanogel by radical graft copolymerization and evaluated its chemo-physical properties (such as, size and zeta potential) using transmission electron microscopy (TEM). According to the principle of charge interaction, the PNIPAM/PEI-siRNA nanogel encapsulated RRM2-siRNA was prepared by conjugating anti-human epidermal growth factor receptor 2 (Her2) antibody. The effect of PNIPAM/PEI-siRNA nanogel complex encapsulated siRNA was determined by agarose gel electrophoresis. The in vitro uptake of NCI-N87 cells by nanogel complex was quantitatively observed by fluorescence microscopy and flow cytometry (FCM). The expression of RRM2 after RRM2 interference using nanogel-siRNA complex in NCI-N87 cells was tested by realtime PCR. The tumor migration suppressing effect of the nanogel complex on Her2-positive tumor cells was determined by Transwell assay. Results The core-shell temperature-responsive PNIPAM/PEI nanogel was synthesized by radical graft copolymerization, with homogeneous size of 359.8 nm, and zeta potential of 21.4 mV. Furthermore, the nanogel complexes encapsulated RRM2-siRNA with different ratios of N/P (N/P ratio: the molar ratio of nitrogen-from-polyethylenimine to phosphate-from-RNA) were prepared and the electrophoresis results showed that the optimal N/P ratio was 60. The cellular uptake experiment showed that the nanogel had good temperature sensitivity and tumor targeting ability at different temperatures (37 ℃, 42 ℃); and the nanogel complex down-regulated the expression of RRM2 and inhibited the migration of NCI-N87 cells. Conclusion The antibody-targeting temperature-sensitive PNIPAM/PEI nanogel is successfully prepared, and it can bind and deliver siRNA into target cells and can be used as a new drug delivery system for anti-tumor therapy.

Abstract:Objective To observe the expression of microRNA-150 (miR-150) in conjunctival mucosa-associated lymphoid tissue (MALT) lymphoma, so as to investigate its mechanism in proliferation, migration and invasion of conjunctival MALT lymphoma. Methods The expressions of miR-150 and Cbl-b, a possible downstream molecule of miR-150, were measured by qPCR in MALT lymphoma tissues and precancerous tissues collected from 3 patients with conjunctival MALT lymphoma in Changzheng Hospital of Second Military Medical University. Then, we transfected miR-150 inhibitor and negative control into human multiple myeloma cell lines RPMI 8226 by cell transfection. CCK-8 assay and flow cytometry (FCM) method were used to investigate the role of miR-150 in the proliferation and apoptosis of RPMI 8226 cells. Transwell assay was used to analyze the effect of miR-150 on the migration and invasion of RPMI 8226 cells. Western blotting analysis was used to examine the regulation of miR150 on Cbl-b expression in RPMI 8226 cells. Results The expression of miR-150 was significantly increased in the conjunctival MALT lymphoma tissues compared with precancerous tissues (P<0.05,P<0.01). Compared with negative control group, the proliferation of RPMI 8226 cells was significantly repressed (P<0.01), the apoptosis was significantly increased (P<0.01), and the migration and invasion were significantly decreased (P<0.05,P<0.01) after transfection of miR-150 inhibitor. The expression of Cbl-b was significantly up-regulated in MALT lymphoma tissues, and was significantly increased after inhibiting miR-150 expression. Conclusion Up-regulated miR-150 can promote the proliferation, migration and invasion of lymphoma cells and is involved in the generation of conjunctival MALT lymphoma, which may be mediated by inhibiting its downstream target gene Cbl-b.

Abstract:Objective To confirm whether fasudil can block C2C12 myoblasts respiration dysfunction triggered by Rho-associated coiled-coil containing protein kinase 1 (ROCK1), and whether it can block the occurrence of muscle atrophy. Methods C2C12 myoblasts were cultured in vitro, and 2% horse serum was used to induce cell differentiation and maturation. The obtained mature muscle tubule cells were divided into four groups according to the different stimuli: Ad-GFP group, only transfected GFP-adenovirus vector (Adv) in C2C12 myoblasts; Ad-ROCK1 group, transfected ROCK1-Adv in C2C12 myoblasts to induce ROCK1 overexpression; Ad-GFPF group, transfected GFP-Adv and given 10 μmol/L fasudil in C2C12 myoblasts; and Ad-ROCK1F group, transfected ROCK1-Adv and given 10 μmol/L fasudil in C2C12 myoblasts. The oxygen consumption rate (OCR) and extracelluar acidification rate (ECAR) of C2C12 myoblasts under different stimulation conditions were evaluated by cell energy metabolism analyzer (Seahorse), so as to determine the effect of ROCK1 overexpression and fasudil stimulation on the respiratory function of C2C12 myoblasts. Mitochondrial fission was measured by MitoTracker^® red fluorescent probes. The expressions of ROCK1, mitochondrial-related protein 1 (Drp1) and phosphorylated p-Drp1, E3 ubiquitin ligase muscle RING finger-1 protein (MuRF1) and muscle atrophy F-box (MAFbx, Atrogin 1) was measured by Western blotting analysis. Results Seahorse analysis showed that the OCR, ECAR, basal respiration, maximal respiration and respiration required for coupling ATP of C2C12 myoblasts in the Ad-ROCK1 group were significantly increased compared with those in the Ad-GFP group (P<0.01); Meanwhile, MitoTracker^® staining showed that the mitochondrial fission was increased and the mitochondrial size frequency distribution shifted left in the Ad-ROCK1 group. After exposed to fasudil, the OCR and EACR of C2C12 myoblasts in the Ad-ROCK1F group were significantly decreased versus the Ad-ROCK1 group, and the basal respiration and maximal respiration were significantly increased (P<0.05). Western blotting analysis showed that p-Drp1/Drp1 ratio, and the expressions of ROCK1, MuRF1 and Atrogin1 in Ad-ROCK1F group were significantly reduced compared with Ad-ROCK1 group (P<0.05). Conclusion Fasudil, an inhibitor of ROCK1, can block the abnormal cell respiration of C2C12 myoblasts caused by overexpressed ROCK1 in vitro, and can reduce the activity of mitochondrial kinetic protein and the expression of muscle atrophy-related proteins.

Abstract:Objective To investigate the protective effect of icaritin liposme against hepatic ischemia /reperfusion (I/R) injury in rats and its mechanisms. Methods A total of 120 male SD rats were randomly divided into four groups: icaritin liposome+I/R (ICT+I/R) group, vechicle liposome+I/R (LIP+I/R) group, I/R group and sham operation (Sham) group. Each group was randomly divided into two subgroups of 2 h and 6 h. The rats in the Sham group were only with free hilum, and the other three groups were subjected to 70% liver ischemia for 60 min. Icaritin liposome (1.5 mg/kg) or vechile lipsome (with same volume of icaritin liposome) were intraportal venously injected in the ICT+I/R and LIP+I/R groups at 10 min before ischemia, without any pretreatment in the I/R group. Blood and liver tissue samples in each group were obtained at 2 h and 6 h after reperfusion to measure the levels of serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST), and the contents of superoxide dismutase (SOD), malondialdehyd (MDA), nitric oxide (NO), nitric oxide synthase (NOS), inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS) and myeloperoxidase (MPO) in liver tissues. The morphology of liver tissues was observed by H-E staining. The apoptosis of liver cells and apoptosis index (AI) were calculated by TUNEL staining. Results Compared with the LIP+I/R and IR groups, ALT level, MDA and MPO contents, and AI were significantly decreased in the ICT+I/R group at 2 h after reperfusion (P<0.05, P<0.01). At 6 h after reperfusion, the ALT and AST levels, MDA content, and AI in the ICT+I/R group were significantly reduced, and the contents of SOD, NO, NOS, and eNOS were significantly increased compared with the LIP+I/R and IR groups (P<0.05, P<0.01). Conclusion Icaritin liposome can reduce liver I/R injury by increasing the contents of SOD and NO, reducing the formation of MDA, promoting the expression of eNOS, and inhibiting the accumulation of MPO and liver cell apoptosis.

Abstract:Objective To investigate the effect of LIM homeobox gene-8 (Lhx8) on the proliferation, metastasis and invasion of human ovarian cancer SKOV3 cells. Methods Lhx8-overexpression lentivirus (LV-Lhx8) was transfected into ovarian cancer SKOV3 cells to establish Lhx8-overexpression model and the negative control lentivirus (LV-NC) was used as control. Then Lhx8-siRNA and FAM-siRNA (used as control; NC group) siRNA was transfected into SKOV3 cells using Lipofectamine 2000 to construct cell interference model. Cells in wild type (WT) group were only given equivalent transfection reagent without virus or any interference fragment. The expression of Lhx8 was detected by immunofluorescence, qPCR and Western blotting. The proliferation of cells after overexpressing or interfering Lhx8 was measured by EDU assays and cell cycle assay. The migration and invasion of cells after transfection were measured by wound scratch experiments and Transwell assay. Results The expression of Lhx8 in SKOV3 cells in the LV-Lhx8 group was significantly higher than that in the LV-NC and WT groups (P<0.01), and its mRNA and protein expressions were significantly decreased after interfering Lhx8 (P<0.01). Compared with the WT and LV-NC groups, the proliferation of SKOV3 cells was significantly decreased in the LV-Lhx8 group and was significantly increased in the Lhx8-siRNA group (P<0.01). The cell cycle assay showed that Lhx8 overexpression significantly inhibited cell proliferation by increasing the number of cells in the G₀/G₁ phase, while the number of cells in the S phase in the Lhx8-siRNA group was significantly higher than that in the WT and NC groups (P<0.01). The migration and invasion of SKOV3 cells and the expression of matrix metalloproteinase (MMP)-2 and MMP-9 in the LV-Lhx8 group were significantly lower than those in the WT and LV-NC groups (P<0.01), while those in the Lhx8-siRNA group were significantly higher (P<0.01). Conclusion Lhx8 can inhibit the proliferation, migration and invasion of ovarian cancer SKOV3 cells, and down-regulate the expressions of MMP-2 and MMP-9.

Abstract:Objective To explore the feasibility of photoplethysmography in monitoring the changes of noxious stimulation intensity caused by veress needle insertion during pediatric laparoscopic hernia repair. Methods Ninety pediatric patients (2-4 years old) with the American Society of Anesthesiologists (ASA) grade ofⅠ-Ⅱ scheduled for laparoscopic hernia repair surgery were randomized into three groups (n=30): group A, B, and C. After tracheal intubation of general anesthesia induction, the pediatric patients in group A received intravenous 0.1 μg/kg sufentanil 5 min before veress needle insertion, group B received 0.1 μg/kg sufentanil immediately after placing the veress needle, while group C received veress needle insertion without sufentanil. The photoplethysmographic amptitude (PPGA) from right forefinger, and surface electromyography (sEMG) of abdominal muscles of pediatric patients were recorded at 6 min prior to veress needle insertion (that is 1 min before receiving sufentanil in the group A, and the injection time was not included, T1), immediately after veress needle insertion (T2), and at 5 min after veress needle insertion (that is 5 min after administration of sufentanil in the group B, T3). Results The values of PPGA had no significant difference between T1, T2 and T3 in group A (P>0.05). Compared with T1 and T3, the value of PPGA at T2 was significantly decreased in group B (P<0.05). The values of PPGA at T2 and T3 were significantly decreased compared with T1 in group C (P<0.05). PPGA was negatively correlated with sEMG in three groups (r=-0.601, -0.512, -0.613; P<0.05). Conclusion Photoplethysmography can quantify the changes of noxious stimulation intensity in pediatric laparoscopic hernia repair and has good application values.

Abstract:Objective To explore the effects of chronic sleep deprivation (CSD) on the ultrastructure and downstream signaling pathway of dopamine D₁ receptor of the hippocampus in rats. Methods Thirty-five male SD rats were selected in this experiment, of which 11 with the lightest weight, shortest weight-bearing swimming time or no finding the platform within 90 s in Morris water maze experiment were excluded. The other 24 rats were randomly divided into tank control (TC) group, CSD group and CSD+dopamine D₁ receptor agonist SKF38393 (SKF) group. The CSD rat model was established by modified multi-platform water environment, and then the rats in the SKF group were intraperitoneally injected with SKF38393 (1 mg/kg) at 15-21 d of CSD. At 21 d after CSD, the ultrastructure of hippocampus was observed by transmission electron microscopy, and the expression of key factors in dopamine D₁ receptor-related signal pathway in the hippocampus was detected by Western blotting and qPCR. Results The mitochondrial swelling, degeneration and destruction of membrane structure of the hippocampus neurons induced by CSD were improved by SKF38393. Compared with the TC group, the mRNA expression levels of adenylate cyclase 5 (Adcy5), protein kinase cAMP-dependent catalytic α (Prkacα), dopamine and cAMP-regulated phosphoprotein (Darpp32), Ras-related protein (Rap) 1a, extracellular signal regulated kinase 1 and 2 (ERK1/2), phospholipase C β1 (PLCβ1), calcium/calmodulin-dependent protein kinase Ⅱa and Ⅳ (CaMKⅡa, CaMKⅣ) in the CSD group were significantly decreased (P<0.05), and the protein levels of total and phosphorylatied protein kinase A catalyzes subunit α (PKAcα), phosphorylated ERK1/2, phosphorylated PLCβ1, and phosphorylated CaMKⅣ were significantly decreased (P<0.05). Compared with the CSD group, the mRNA expressions of Prkacα, Darpp32, Rap1a, Rap1b, ERK1 and CaMKⅣ in the SKF group were significantly increased (P<0.05), and the protein levels of total and phosphorylated PKAcα and phosphorylated CaMKⅣ were significantly increased (P<0.05), while the expression of PLCβ1 and total CaMKⅣ was similar in the two groups. Conclusion CSD damages the ultrastructure of the hippocampus neurons in rats, which can be effectively improved by dopamine D₁ receptor agonist SKF38393, and the protective mechanism may be related to the PKA pathway and phosphoinositol pathway.

Abstract:Objective To investigate the spatial distribution, cell localization and time-dependent changes of 5-lipoxygenase (5-LOX) expression in brain tissues of rats with surgical brain injury (SBI) and its role in the pathogenesis of SBI. Methods Seventy-two healthy male SD rats were randomly divided into Sham group, 1-day post surgery (SBI-1d) group, 3-day post surgery (SBI-3d) group and 7-day post surgery (SBI-7d) group, each group with 18 rats. SBI rat model was established by right frontal lobectomy in the SBI-1d, SBI-3d and SBI-7d groups, while rats in the Sham group only with the corresponding skull removed with the dura intact. Brain water content (BWC) of ipsilateral and contralateral brain tissues was measured by wet-dry weight formula. The neurobehavioral functions of all rats were evaluated by modified Garcia score and beam balance test. The spatial distribution and cellular location of 5-LOX were detected by immunofluorescence staining. The expressions of 5-LOX and NF-κB in the damaged brain tissues were detected by Western blotting analysis. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) around the lesion areas were determined by biochemical method. Results (1) Compared with the Sham group, neurological dysfunction was significant in the SBI-1d, and SBI-3d and SBI-7d groups (P<0.01), and the BWC of injured brain tissue of rats was significantly increased in the SBI-1d and SBI-3d groups (P<0.05). The modified Garcia score in the SBI-7d group was significantly higher than that in the SBI-1d and SBI-3d groups (P<0.05). (2) 5-LOX was mainly distributed around the lesion areas, which was mainly localized in the cytoplasm of neurons, followed by glial cells and microglia. (3) The expressions of 5-LOX and NF-κB were significantly increased in the SBI-1d and SBI-3d groups (P<0.05) versus the Sham and SBI-7d groups. (4) Compared with the Sham group, the activity of SOD was significantly decreased in the other three groups (P<0.05); while the content of MDA was significantly increased in the SBI-1d and SBI-3d groups (P<0.05). Conclusion 5-LOX is mainly expressed in the neurons around the lesion areas after SBI, followed by glial cells and microglia, with the highest expression at 1 day after surgery. The mechanism by which 5-LOX aggravates brain injury may be related to increased expression of NF-κB and oxidative stress injury.

Abstract:Protein phosphatases play critical roles in regulating cell division, cell apoptosis, and cell cycle in eukaryotic cells, participating in numerous signal transduction processes and exerting a large amount of significant biological functions. A variety of protein phosphatases are identified to maintian the phosphorylation level of key proteins with a moderate level in the antiviral innate immune response to virus infection. In this paper, we reviewed the roles of serine/threonine protein phosphatases, tyrosine protein phosphatases, and lipid phosphatases in regulating antiviral innate immune responses.

Abstract:Advances in biotechnology give much importance to the therapeutic biomacromolecules in the therapy of diseases, such as proteins, oligonucleotides, and peptides. But their effects are limited in practical application because of cell membrane barrier. Cell-penetrating peptides (CPPs) are promising oligopeptides with a remarkable capacity for membrane translocation, which can carry various macromolecules into cells. In this paper, we reviewed the classification and transmembrane mechanism of CPPs as nanoparticles, with particular focus on their recent progress in tumor-targeted therapy.

Abstract:Bile acids, as one of the constituents of bile, can promote the digestion and absorption of fat and cholesterol metabolism. Studies find that bile acid is a signal molecule that not only combines with nuclear hormone receptors such as farnesoid X receptor, and G protein coupled bile acid receptor 5, to regulate their own metabolism, but also plays an important role in diabetes, obesity, non-alcoholic fatty liver disease and other metabolic diseases. The research on its mechanism will provide a solid theoretical basis for the diagnosis and treatment of metabolic diseases. In this review, we summarized the research progress of bile acids in glucolipid metabolic diseases.

Abstract:Objective To explore the clinical efficacy of proprioceptive neuromuscular facilitation (PNF) and acupuncture in the treatment of naval soldiers with stiff neck. Methods Fifty naval soldiers with neck stiffness were divided into PNF group and acupuncture group (received PNF or acupuncture treatment, respectively), each group with 25 cases. The Visual Analogue Scale (VAS) score of patients in the two groups was compared immediately and at 3 months after treatment, the degree of range of motion was compared immediately after treatment, and the recurrence rate was compared at 5 months after treatment. Results The VAS scores of patients in the two groups immediately after treatment were significantly lower than those before treatment (1.9±0.5 vs 7.6±1.6, 2.7±0.7 vs 8.0±1.4; both P<0.01). The VAS score in the PNF group was significantly higher than that in the acupuncture group immediately after treatment (2.7±0.7 vs 1.9±0.5, P<0.05), with no significant difference at 3 months after treatment. The range of motion in all directions of patients in the acupuncture group immediately after treatment were significantly improved versus those before treatment (P<0.01), and were significantly better than those in the PNF group (P<0.01). The recurrence rate in the PNF group (4.2%, 1/24) was lower than that in the acupuncture group (25.0%, 6/24), but the difference was not significant (P>0.05). Conclusion Acupuncture has a better immediate effect than PNF in treating patients with stiff neck, but PNF is simple to operate and may reduce the recurrence rate of stiff neck, indicating that PNF can be used as an alternative in the treatment of stiff neck.

Abstract:Objective To analyze the epidemiological changes, clinical features and disease patterns of 10-year renal biopsy series in our center and to compare the changes of pathological diagnosis spectrum of renal biopsy, so as to explore the trend of epidemiological characteristics of renal biopsy across ten years. Methods From January 2007 to December 2016, 1 786 patients (aged ≥18 years) undergoing percutaneous renal biopsy were involved and divided into 3 period groups according to the date of biopsy, earlier group (2007-2010), mid-term group (2011-2013) and recent group (2014-2016). The patients were also divided into 3 age groups (15-39 years, 40-64 years and ≥65 years). The clinical and pathological data of all patients were collected, and than statistical analysis was performed by SPSS 18.0 software. Results A total of 1 786 cases with complete clinical data were enrolled, of which 973 were male and 813 were female, with a median age of 42 years (range 15-88 years). There were 1 548 patients with primary glomerular disease, with IgA nephropathy (IgAN) being the most common one, accounting for 29.1% (520/1 786), 17.3% (309/1 786) having minimal change disease (MCD), and 17.0%(304/1 786) having membranous nephropathy (MN). Totally 238 patients had secondary renal disease, of which lupus nephritis was the most common one, accounting for 3.4% (61/1 786), followed by diabetic nephropathy (2.9%, 51/1786) and Henoch-Schnlein purpura nephritis (2.2% 39/1 786). There were no significant differences in gender or age between three period groups. Compared with the earlier group (66/684, 9.6%), the proportions of MN in the midrtem group (90/547, 16.5%) and the recent group (148/555, 26.7%) were significantly increased (P<0.01). Nephritis syndrome (NS; 45.5%, 812/1786) was the most frequent clinical manifestation in all cases, followed by nephrotic syndrome (688/1786, 38.5%). The most common clinical manifestation in patients suffered from NS was MCD in 15-39 years group and MN in 40-64 years group and ≥65 years group. Meanwhile, the most common diagnosis for patients suffered from nephritis syndrome was IgAN in 15-39 years group and 40-64 years group, but was MN in ≥65 years group. Conclusion IgAN remains the most common glomerulopathy in our study. However, the prevalence of MN has grown quickly in recent years. IgAN is the main diagnosis for young and middle aged patients with nephritis syndrome, while the MN is the main for middle aged and elderly patients with NS.

Abstract:Objective To evaluate the efficacy and safety of tacrolimus (TAC) therapy in patients with refractory IgA nephropathy. Methods Nine IgA nephropathy patients were included in this study were treated from Jun. 2008 to Sep. 2013 in Changzheng Hospital of Second Military Medical University. All patients received TAC therapy after the renin-angiotensin system (RAS) blockade therapy and steroid therapy failed. The main outcome was complete or partial remission. Secondary outcomes included the time required to remission, the frequency of recurrence, TAC dosage and adverse events. Results The initial dosage of TAC was (1.89±0.33) mg/d. After treatment with TAC for 6 months, 6 patients achieved complete remission, 2 partial remission and 1 treatment resistance, and most of the remission patients achieved remission during the first 2 months of TAC therapy. The urine protein level of enrolled patients was significantly decreased ([3.05±1.35] g/24 h vs [0.85±1.54] g/24 h, P<0.05) and the serum album level of all patients was significantly improved ([27.00±8.37] g/L vs [37.33±8.08] g/L, P<0.05). One patient receiving TAC therapy presented worsened hypertension, and no other adverse event was observed in this study. Three of 8 proteinuria remission patients had relapses and achieved remission by adjusting the dosages of steroids and tacrolimus. Conclusion TAC can improve proteinuria in patients with refractory IgA nephropathy, with less adverse reactions.

Abstract:Objective To develop a new type of anti-fatigue nutrient chewable tablet and to explore its anti-fatigue effect in mice. Methods Ginseng polysaccharide, acanthopanax extract and taurine were mixed together in a proportion of 6:8:1 as remedium cardinale, mannitol was used as filler, sucrose as a correctant and magnesium stearate as a lubricant. Hardness, appearance and taste were utilized as score indexes, the dosage of mannitol, sucrose, and magnesium stearate were used as study factors. The orthogonal test was applied to screen the best proportion of raw materials to create the formulation according to the mouse behaviors. A total of 60 male mice were randomly divided into control group (normal saline) and three doses of Shenjia chewable tablet groups (200, 600 and 1 800 mg/kg). Lavage administration was lasted for 7 days. We then carried out the weight loading swimming test to record the swimming time of mice, and also recorded the body mass and the serum biochemical indexes, including adenosine triphosphate (ATP), lactic acid (Lac), free triiodothyronine (FT3), free thyroid hormone (FT4), cortisol and melatonin (MT). Results The formulation was optimized as 30% ginseng polysaccharide, 40% acanthopanax extract, 5% taurine, 19% mannitol, 5% sucrose, and 1.0% magnesium stearate. The Shenjia chewable tablet tasted cool and sweet, round in shape, yellow in color and smooth in appearance. Compared with the control group, there were significant increases in the loaded-swimming time in the three doses groups (P<0.01), in the levels of ATP, FT3, FT4 and MT in middle and high dosage groups (P<0.05, P<0.01), and in Lac level in high dosage group (P<0.05). Cortisol was increased in the three doses groups (P<0.05, P<0.01). All mice in three doses groups had no significant increase in body weight. Conclusion The Shenjia chewable tablet is an ideal anti-fatigue nutrient with reasonable formulation, simple preparation technology, high drug loading and good taste.

Abstract:Objective To explore the effect of Shuxuening on early myocardial injury markers and oxidative stress in severe sepsis patients with myocardial injury. Methods A total of 102 severe sepsis patients with myocardial injury, who received therapy in our hospital from Mar. 2013 to Jun. 2016, were randomly divided into two groups: control group (51 cases) and therapy group (51 cases). The patients in the control group received routine treatments and the therapy group were given Shuxuening based on routine treatment. We detected the levels of plasma N-terminal pro-brain natriuretic peptide (NT-proBNP), heart-type fatty acid binding protein (hFABP), cardiac troponin I (cTnI), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of patients before treatment (at admission), after treatment for 6 h and 72 h (after admission), and analyzed the change of left ventricular ejection fraction (LVEF), acute physiology, chronic health evaluation (APACHE) Ⅱ score, length of ICU stay and 28-day mortality of the patients. Results At admission, there was no difference in the levels of plasma NT-proBNP, hFABP, cTnI, MDA, SOD, GSH-Px, APACHE Ⅱ score or LVEF between the two groups (P>0.05). After treatment for 6 h, the levels of plasma NT-proBNP, hFABP, cTnI and MDA in the therapy group were significantly lower and the activities of plasma SOD and GSH-Px were significantly higher than those in the control group (P<0.05); the two groups both had significant increases in the levels of NT-proBNP, hFABP, cTnI and MDA and significant decreases in the activities of SOD and GSH-Px compared with those before treatment (P<0.05). After treatment for 72 h in the two groups, the levels of plasma hFABP and MDA were significantly lower and the levels of plasma NT-proBNP and cTnI, the activities of SOD and GSH-Px were significantly higher than those after treatment for 6 h (P<0.05). After treatment for 72 h, the levels of plasma NT-proBNP, cTnI and MDA in control group were significantly higher and the activities of plasma SOD and GSH-Px were significantly lower than those in the therapy group (P<0.05), with the difference in the hFABP levels being similar in the two groups (P>0.05). APACHE Ⅱ scores after treatment were significantly lower than that before treatment in the two groups (P<0.05), and no significant difference was found in the LVEF or the length of ICU stay after treatment between the two groups (P>0.05). The therapy group had a significantly lower 28-day mortality than the control group (P<0.05).The therapy group had a significantly lower mortality than the control group (P<0.05). Conclusion Therapeutic effect of Shuxuening on the severe sepsis patients with myocardial injury is superior to the routine treatment. Shuxuening can decrease the levels of NT-proBNP, hFABP, cTnI, and MDA and APACHE Ⅱscore, and can increase the activities of SOD and GSH-Px.

Abstract:Objective To evaluate the effect of topical olive oil, vaseline cream and menthol ointment on skin barrier function of patients with senile xerosis. Methods Totally 32 participants with senile xerosis were enrolled in this randomized double-blind self-controlled study for 8 weeks. Four black square spaces were marked on bilateral forearms of each volunteer, and three were treated with olive oil, vaseline cream and menthol ointment, respectively, once a day for 8 weeks, and the last one was taken as control. Skin barrier function was evaluated at 2, 4, and 8 weeks after experiment by measuring the water content of the stratum corneum, transepidermal water loss (TEWL), and skin pH value. Results Compared with the control group, the water contents of stratum corneum in the olive oil, vaseline cream and menthol ointment groups were significantly increased (P<0.05), and the TEWL value and skin pH value were significantly decreased (P<0.05) at 8 weeks. There were no significant difference in the water content of stratum corneum, TEWL value, or pH value between 3 experiment groups. Conclusion Topical olive oil can improve the skin barrier function of patients with senile xerosis, and vaseline cream and menthol ointment also can repair the skin barrier function.

Abstract:Objective To screen the differentially expressed microRNAs (miRNAs) in colorectal cancer tissues with high expression of gastrin (GAS) by gene chip technique. Methods The level of GAS in tumor tissues from 71 patients with colorectal cancer was detected by enzyme linked immumosorbent assay (ELISA), and the relationship between positive expression of GAS and clinicopathological features was analyzed. Human colorectal cancer tissues with high positive GAS expression (high-expression group, n=4) or negative GAS expression (control group, n=4) were analyzed by miRNA microarray, and the selected differentially expressed miRNAs were validated by qPCR. Results There were 17 cases (23.9%) with GAS content ≥50.00 pg/g (positive GAS expression), while the remaining 54 cases (76.1%) had negative GAS expression. The expression level of GAS was related to the degree of differentiation, Dukes stage and histological type (P<0.05). There were 236 miRNAs had significant differential expression in the high-expression group (GAS content >200.00 pg/g) compared with the control group, of which 159 miRNAs were up-regulated and 77 were down-regulated. A total of 24 miRNAs were screened out, which differentially expressed more than 3 folds in the high-expression group, with 17 up-regulated and 7 down-regulated. Three up-regulated or down-regulated miRNAs were selected for qPCR verification, and the results were consistent with the microarray analysis. Conclusion The miRNAs are differentially expressed in the colorectal cancer tissues with high expression of GAS, and they would be potential targets for the treatment of the colorectal cancer with high GAS expression.

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