Network pharmacological analysis and experimental study of Pulsatilla chinensis against inflammatory injury caused by pneumonia in mice infected with influenza virus FM_1 / 中国中药杂志

Network pharmacology and the mouse model of viral pneumonia caused by influenza virus FM_1 were employed to explore the main active components and the mechanism of Pulsatilla chinensis against the inflammatory injury of influenza virus-induced pneumonia. The components and targets of P. chinensis were searched from TCMSP, and the targets associated with influenza virus-induced pneumonia were searched from GeneCards. The common targets between P. chinensis and influenza virus-induced pneumonia were identified with Venn diagram established in Venny 2.1. The herb-component-disease-target(H-C-D-T) network was constructed by Cytoscape 3.7.2. The above data were imported into STRING for PPI network analysis. Gene Ontology(GO) enrichment and KEGG pathway enrichment were performed with DAVID. BALB/cAnN mice were infected with the influenza virus FM_1 by nasal drip to gene-rate the mouse model of pneumonia. Immunohistochemistry was adopted to the expression profiling of inflammatory cytokines in the lung tissues of mice in the blank group, model group, and P. chinensis group 1, 3, 5, and 7 days after infection. The pathological changes of lung and trachea of mice in blank group, model group, and P. chinensis group were observed with light microscope and scanning electron microscope at all the time points. The network pharmacological analysis indicated that 9 compounds of P. chinensis were screened out, with a total of 57 targets, 22 of which were overlapped with those of influenza virus-induced pneumonia. A total of 112 GO terms(P<0.05) were enriched, including 81 terms of biological processes, 11 terms of cell components, and 20 terms of molecular functions. A total of 53 KEGG signaling pathways(P<0.05) were enriched, including TNF signaling pathway, influenza A signaling pathway, NF-κB signaling pathway, MAPK signaling pathway and other signaling pathways related to influenza/inflammation. In the P. chinensis group, the expression of TNF-α and IL-1 in the lung tissue was down-regulated on the 3 rd day after infection, and that of IL-6 in the lung tissue was down-regulated on the 5 th day after infection. Light microscopy and scanning electron microscopy showed that P. chinensis significantly alleviated the pathological damage of lung and trachea compared with the model group. This study reflects the multi-components, multi-targets, and multi-pathways of P. chinensis against influenza virus-induced pneumonia. P. chinensis may reduce the production of proinflammatory cytokines and mediators and block the pro-inflammatory signaling pathways to alleviate viral pneumonia, which provides reference for future research.

Surveillance of adverse events following immunization with measles-containing vaccines in Shanghai， 2011-2018 / 上海预防医学

Objective:To analyze the safety of measles-containing vaccines （MCV） in Shanghai city. Methods:Adverse event following immunization cases related to measles attenuated live vaccine （MV）， measles and rubella combined attenuated live vaccine （MR） and measles， mumps and rubella combined attenuated live vaccine （MMR） vaccination in Shanghai during 2011—2018 were collected from the National AEFI Surveillance System for epidemiological analysis. Results:A total of 15 055 MVC-related AEFI cases were reported， for an incidence of 139.74 per 100 000 doses. Among all MVC-related AEFI cases， the male to female ratio was 1.17∶1， and 12 068 （111.70 per 100 000 doses） were common reactions， with 1 050 from MV（64.66 per 100 000）， 5 484 from MR（134.36 per 100 000）and 5 534 from MMR（108.60 per 100 000. Most of common reactions were fever （96.01 per 100 000）， but 2 939 （27.20 per 100 000） were rare reactions， with 408 from MV （25.12 per 100 000）， 1 983 from MR （48.59 per 100 000） and 548 from MMR （10.75 per 100 000）. Most of rare reactions were allergic rash （25.00 per 100 000）. Conclusion:MCV has a good safety profile but we should continue to strengthen MCV-related AEFI surveillance.

Expert consensus on Antiviral Oral Liquid in treatment of influenza in clinical practice / 中国中药杂志

Antiviral Oral Liquid is modified on the basis of Baihu Decoction in Treatise on Febrility Diseases by ZHANG Zhongjing and Qingwen Baidu Yin in Qing Dynasty, with effects in clearing toxic heat, repelling dampness and cooling blood. It is widely used in clinical treatment of common colds, influenza and upper respiratory tract infection, mumps, viral conjunctivitis and hand-foot-mouth disease, with a good clinical efficacy and safety. Based on a questionnaire survey of clinicians and a systematic review of study literatures on Antiviral Oral Liquid, the international clinical practice guidelines development method was adopted to analyze the optimal available evidences and expert experiences in the "evidence-based, consensus-based and experience-based" principles. The consensus was jointly reached by more than 30 multidisciplinary experts nationwide, including clinical experts of traditional Chinese and Western medicine in the field of respiratory diseases and infectious diseases, and methodological experts. In the study, literatures were retrieved based on clinical problems in the clinical survey as well as PICO clinical problems. The GRADE system was used for the classification and evaluation of evidence, and fully combined with clinical expert experience, so as to reach expert consensus by the nominal grouping method. This expert consensus recommended or suggested indications, usage and dosage, course of treatment, intervention time for treatment, and the safety and precautions of Antiviral Oral Liquid for treatment of influenza, and can provide reference for the rational use of this drug in clinical practice.

Current quality standards of cattle bile and sheep bile quality standards and discussion on related problems / 中国中药杂志

To analyze quality standards of cattle bile and sheep bile, and to discuss the related problems in the standards. The results showed that physical forms of the related medicinal materials of cattle bile and sheep bile were chaotic, and the technical methods adopted in the quality standards were generally backward. In addition, there were still problems that some medicinal material standards lacked necessary test items, which were especially obvious in the relevant medicinal material standards of sheep bile and brought difficulties to quality evaluation and control. We suggest that physical forms of cattle bile and sheep bile in quality standards should be determined, and inspection items should be completed. Based on mainstream analytical technology, some technical methods of these standards should be improved.

Effects of nanofibers on mesenchymal stem cells: environmental factors affecting cell adhesion and osteogenic differentiation and their mechanisms / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Nanofibers can mimic natural tissue structure by creating a more suitable environment for cells to grow, prompting a wide application of nanofiber materials. In this review, we include relevant studies and characterize the effect of nanofibers on mesenchymal stem cells, as well as factors that affect cell adhesion and osteogenic differentiation. We hypothesize that the process of bone regeneration in vitro is similar to bone formation and healing in vivo, and the closer nanofibers or nanofibrous scaffolds are to natural bone tissue, the better the bone regeneration process will be. In general, cells cultured on nanofibers have a similar gene expression pattern and osteogenic behavior as cells induced by osteogenic supplements in vitro. Genes involved in cell adhesion (focal adhesion kinase (FAK)), cytoskeletal organization, and osteogenic pathways (transforming growth factor-β (TGF-β)/bone morphogenic protein (BMP), mitogen-activated protein kinase (MAPK), and Wnt) are upregulated successively. Cell adhesion and osteogenesis may be influenced by several factors. Nanofibers possess certain physical properties including favorable hydrophilicity, porosity, and swelling properties that promote cell adhesion and growth. Moreover, nanofiber stiffness plays a vital role in cell fate, as cell recruitment for osteogenesis tends to be better on stiffer scaffolds, with associated signaling pathways of integrin and Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ). Also, hierarchically aligned nanofibers, as well as their combination with functional additives (growth factors, HA particles, etc.), contribute to osteogenesis and bone regeneration. In summary, previous studies have indicated that upon sensing the stiffness of the nanofibrous environment as well as its other characteristics, stem cells change their shape and tension accordingly, regulating downstream pathways followed by adhesion to nanofibers to contribute to osteogenesis. However, additional experiments are needed to identify major signaling pathways in the bone regeneration process, and also to fully investigate its supportive role in fabricating or designing the optimum tissue-mimicking nanofibrous scaffolds.

Analysis of pathogenesis clauses and prescriptions of “Treatise on differentiation and treatment of epidemic febrile disease” related to clinical symptoms of COVID-19 / 中草药

Objective: To collect the clinical symptoms of novel coronavirus pneumonia, explore and analyze the clauses and prescriptions related to the clinical symptoms of 2019 novel coronavirus pneumonia in “Treatise on differentiation and treatment of epidemic febrile disease”, in order to provide a reference for clinical treatments of the disease. Methods: Searching literature from the databases CNKI and PubMed by entering “novel coronavirus” (in Chinese) and “2019 Novel Coronavirus” as key words, classifying typical symptoms depending on their reported frequencies of occurrence, comparing with related pathogenesis in “Treatise on differentiation and treatment of epidemic febrile disease”, and analyzing clauses and prescriptions combined with the sixth version of “New coronavirus pneumonia diagnosis and treatment scheme”. Results: We optimized 20 literatures published in Chinese or English for the review, in which, a total of 647 clinical cases were reported. Based on that, 28 typical symptoms were summarized, including respiratory symptoms, such as fever and cough; Gastrointestinal symptoms, such as poor appetite, nausea, vomiting, diarrhea; Other symptoms, such as myalgia, fatigue, and so forth. Among them, fever (558 cases), cough (395 cases), and fatigue (257 cases) were the top three clinical symptoms. By analysis, we optimized 15 clauses, 18 prescriptions, and 46 traditional Chinese medicines from “Treatise on differentiation and treatment of epidemic febrile disease”, including two clauses in the upper energizer, nine in the middle energizer, and four in the lower energizer; A total of seven prescriptions for treating dampness-heat, five for cold-dampness, and six for warm-heat diseases; as well as Chinese medicines Pinellia ternata (mentioned for six times), Cinnamomi Ramulus (five times), Tetrapanacis Medulla (five times), Poria cocos (four times), Talcum (four times), Coptis chinensis (four times) and Coicis Semen (four times), etc. P. ternata, Cinnamomi Ramulus, C. chinensis, Scutellaria baicalensis, and Zingiberis Rhizoma were selected to treat dampness-heat diseases, in order to “pungency opening and bitter discharging”; P. cocos, Tetrapanacis Medulla, Talcum, and Coicis Semen were selected for “eliminating dampness with bland medicinal”; Almond, Amomum kravanh, and Magnolia officinalis were used to “regulating qi movement”; Anemarrhenae Rhizoma, Gypsum Fibrosum, and Forsythiae Fructus were used to treat warm-hot diseases for “purging fire and removing toxin”. The formula of Sanren Decoction, Yiyizhuye Powder can be used for people in the stage of medical observation period; Sanren Decoction, Yiyizhuye Powder, Xingrenyiyi Decoction, Wuling Powder, Baihujiaguizhi Decoction, and Lingjiangzhugui Decoction for treating mild cases; Huangqin Huashi Tang, Xingren Huashi Decoction, and Fulingpi Decoction for severe cases; Jiaomei Decoction, Jianweiwumei Pill, Sini Decoction, and Taohua Porridge for critical patients, and Maidong Maren Decoction, Wuzhi Yin, and Niuru Yin for convalescents. Conclusion: The prescriptions of “Treatise on differentiation and treatment of epidemic febrile disease” can provide reference for the clinical treatments of 2019 novel coronavirus pneumonia.

Application Status of Chinese Medicine on Cancer Rehabilitation: A Preliminary Questionnaire Survey / 中国结合医学杂志

OBJECTIVE@#To investigate the application status of Chinese medicine (CM) in cancer rehabilitation, so as to provide references for improving the level of CM cancer rehabilitation.@*METHODS@#A survey questionnaire regarding "application status of CM rehabilitation in cancer patients" for oncologists (doctor version) and cancer patients (patient version) were developed, respectively. From September 2014 to November 2016, a total of 1,000 doctors from oncology department in 48 hospitals and 2,000 cancer patients from CM oncology department from 8 hospitals in China were recruited in this survey. The psychological, nutrition and exercise rehabilitation guidance for cancer patients provided by doctors, their mastery conditions regarding cancer staging treatment and CM syndrome differentiation, and recommendation from doctors on CM rehabilitation were investigated. Cancer patients' awareness on the importance of psychological, nutrition and exercise rehabilitation, as well as their knowledge and needs for rehabilitation were also analyzed. The impact of gender, age, professional title, hospitals grades of physicians on their knowledge of cancer staging treatment and CM syndrome differentiation, and the relationship between gender, age, education level and economic conditions and patient's knowledge along with the needs of CM rehabilitation were further analyzed.@*RESULTS@#Totally 1,000 questionnaires were issued to doctors and 963 questionnaires returned, among which 948 were valid representing a response rate of 94.80%. A total of 2,000 questionnaires were issued to patients and 1,705 valid data finally returned with a response rate of 85.25%. The survey showed that cancer patients generally paid much attention to psychological, nutritional and sports rehabilitation, and had a strong demand for CM rehabilitation. Knowledge of CM rehabilitation was not well provided by oncologists, and the rehabilitation guidance as well as CM rehabilitation measures were obviously insufficient in cancer patients. Educational and economic levels were positively correlated with cognition level of CM rehabilitation knowledge among cancer patients (Kendall-tau_b correlation coefficients=0.130, 0.057, respectively; P<0.05). Gender and education level were positively correlated with the patients' willingness for taking CM measures (Kendall-tau_b correlation coefficient=0.057, 0.105, respectively; P<0.05). Age was negatively correlated with intention of applying CM measures (kendall-taub correlation coefficient=-0.105, P<0.05).@*CONCLUSIONS@#Health education and professional training for both cancer patients and oncologists should be strengthened and CM rehabilitation knowledge among cancer patients and oncologists should be improved, so as to give full play to CM in cancer rehabilitation.

Correlation of serum C-telopeptide cross-linked collagen type Ⅰ with bone mineraI density in a rat modeI of disuse osteoporosis / 中国组织工程研究

BACKGROUND: C-telopeptide and N-telopeptide cross-linked collagen type Ⅰ (CTx and NTx, respectively) are specific biochemical bone markers that can reflect bone formation and resorption. OBJECTIVE: To analyze the association of CTx with disuse osteoporosis. METHODS: Male Sprague-Dawley rats, weighing 180-220 g, were randomly divided into control and disuse osteoporosis groups. Right hind limbs of the rats in the disuse osteoporosis group were immobilitzed for 4 weeks by ankle-tail fixation to establish the rat model of disuse osteoporosis. Peritoneal venous blood was collected before and after modeling, and the femur was then removed to measure the serum CTx level and bone mineral density of the bilateral femurs. RESULTS AND CONCLUSION: The serum CTx level did not differ significantly between groups before modeling (P > 0.05). At 4 weeks after modeling, the serum CTx level in the disuse osteoporosis group was significantly higher than that in the control group and at baseline (P <0.01). The serum CTx level showed no significant change in the control group before and after modeling (P > 0.05). The increment of serum CTx in the disuse osteoporosis group exhibited a negative correlation with the bone mineral density of the bilateral femurs (r=0.426, P < 0.01). The bone mineral density of the right femur in the disuse osteoporosis group was significantly lower than that of the left one in the disuse osteoporosis group and the right one in the control group (P < 0.01), and there was no significant difference in the bone mineral density between left and right femurs in the control group (P > 0.05). These results imply that the model of disuse osteoporosis by ankle-tail fixation is established successfully. Disuse osteoporosis can promote the production of CTx further reducing bone mineral density; CTx is positively correlated with the degree of bone loss, so it can be used for therapeutic assessment and diagnosis of osteoporosis.

Intervention effect of CD147 on learning and memory ability in rat model of Alzheimer's disease / 天津医药

Objective To investigate the intervention effect of CD147 on learning and memory ability in rat model of Alzheimer's disease. Methods A total of 60 healthy Sprague-Dawley rats were randomly divided into sham operation group, model group and CD147 group, 20 rats in each group. All of the rats were anesthetized with intraperitoneal injection of 10％chloral hydrate (0.3 g/kg). The rats in the model group and the CD147 group were injected with Aβ1-40 (10μg) in the bilateral hippocampal CA1 regions, while the rats in the sham operation group were injected with the same amount of saline at the same sites. After 48 h, the rats in CD147 group were injected with CD147 cDNA in the bilateral ventricles, while the rats in model group and sham operation group were injected the same amount of saline at the same sites. Morris water maze test was used to detect the ability of learning and memory of rats. The expressions ofβamyloid protein (Aβ) andγ-secretase were detected by Western blot assay. Results The escape latency was significantly longer in model group than that of sham operation group, while which was significantly lower in CD147 group than that of model group (P<0.05). The number of times across the platform and the time of staying on platform were significantly lower in model group than those of sham operation group, while which was significantly higher in the CD147 group than that of model group (P<0.05). The expressions of Aβandγ-secretase were increased significantly in model group compared to those of sham operation group, while which were significantly decreased in CD147 group compared with those of model group (P<0.05). Conclusion Exogenous CD147 can significantly improve the learning and memory ability of AD rats, and its specific mechanism may be related to regulating the activity ofγ-secretase and down regulating the expression of Aβ.

Effects of Yiqi Huayu Jiedu Prescription on MHCC97-H Migration Ability and Expressions of Chemokines CXCL12, CXCR4 and CXCR7 / 中国中医药信息杂志

Objective To study the effects of Yiqi Huayu Jiedu Prescription on the migration ability of MHCC97-H and the expressions of CXCL12, CXCR4 and CXCR7; To discuss its relevant mechanism of action. Methods Setting Sorafenib as a positive control, CCK-8 method was used for determining the effects of Yiqi Huayu Jiedu Prescription on the cell proliferation of MHCC97-H and the optimum concentration. Scratch assay was used to observe the migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 were detected by Western blot after 24 h of medicine intervention. Results Yiqi Huayu Jiedu Prescription and Sorafenib can inhibit the cell proliferation of MHCC97-H , and the inhibitory concentration was 0.095 g/mL and 10 μmol/mL at 24 hours. Yiqi Huayu Jiedu Prescription can inhibit migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 in hepatocellular carcinoma cells decreased after the action of Yiqi Huayu Jiedu Prescription. Conclusion Yiqi Huayu Jiedu Prescription can inhibit MHCC97-H cell proliferation and migration, which may be realized by down-regulating chemokine axis of CXCL12/CXCR4/CXCR7.

Role of up-regulation of THEM4 on extracellular matrix accumulation in kidney of diabetic mice / 中国药理学通报

Aim To observe the effect of thioesterase superfamily member 4 ( THEM4 ) expression on extra-cellular matrix ( ECM ) accumulation in the kidney of diabetic mice .Methods For in vivo vector delivery experiment , male CD1 mice were randomly divided in-to four groups: normal control mice ( Control group ) , diabetic mice ( DM group ) , diabetic mice receiving pYr-ads-4-THEM4 vector ( DM+THEM4 vector ) and diabetic mice receiving pYr-adshuttle-4 vector ( DM+V vector ) .pYr-ads-4-THEM4 vector or pYr-adshuttle-4 vector ( 1 mg · kg -1 ) were mixed with TransIT-EE Hydrodynamic Delivery Solution from Mirus Co .and injected into tail vein once a week for four weeks after STZ injection.Four weeks later, mice were sacrificed and Western blot , immunohistochemistry and real-time PCR were used to detect the expression of phospho-Akt(Ser 473), THEM4, TGF-β1, α-SMA, Col Ⅲ, FN proteins and THEM4 mRNA in kidneys respectively . Results THEM4 decreased in kidney of diabetes mel-litus accompanied with increased phospho-Akt ( Ser 473), TGF-β1, α-SMA and ECM.The delivery of pYr-ads-4-THEM4 vector increased THEM4 expression and decreased phospho-Akt (Ser 473), TGF-β1, α-SMA and ECM deposit in kidneys of diabetic mice . Conclusion The up-regulation of THEM4 may pre-vent ECM deposit by inhibiting the phosphorylation of Akt and down-regulating the expression of TGF-β1 andα-SMA in kidneys of diabetic mice .

Content Determination of As and Hg of Lilium Lancifolium Samples from Longshan County by AFS / 中国中医药信息杂志

Objective To analyze and discuss the content determination of As and Hg from Lilium Lancifolium in Longshan County; To optimize microwave digestion conditions. Methods Automatic microwave digestion appratus was used. Lilium Lancifolium samples from Longshan County were digested in teflon microwave tube with HNO3-H2O2, and As and Hg were measured with atomic fluorescence spectrometry (AFS). Results The content of As was in the range of 0.031–0.507 mg/kg, and the highest content of Hg was 0.024 mg/kg. The regression equation was Y=221.23X+170.72(r=0.995 9),Y=503.52X-682.43,(r=0.999 2).For the production base,the recoveries of As and Hg were 94.32% and 92.48% in the samples, and RSD were 2.14% and 2.70%; for the breeding base, the recoveries of As and Hg were 94.95% and 93.52% in the samples, and RSD were 1.15% and 1.97%. Conclusion The method is simple and reliable, which can be used to the content determination of As and Hg from Lilium Lancifolium, and provide references for the choice of base of production and breeding of Lilium Lancifolium in Longshan County.

Effects of Qingjin Granules on Methicillin-resistant Staphylococcus Aureus Pneumonia Mice / 中国中医药信息杂志

Objective To observe the effects of Qingjin Granules on methicillin-resistant staphylococcus aureus (MRSA) pneumonia mice. Methods Pneumonia model was prepared by intranasal drip of bacterial fluid. Experimental mice were randomly divided into control group, model group, Western medicine group and TCM group. On the third day after infection, the Western medicine group was treated with linezolid, and the TCM group was given Qingjin Granules for gavage. The control and model group were given the same amount of normal saline for gavage. Body weight and clinical manifestations every day of each mouse after infected were recorded. Lung scans at day 3 and day 10 were taken; the pathological changes trough HE staining were observed; microbial load of lung tissue was detected. Results There were significant weight losses on mice except control group on the first day after infection. CT scan showed that the lung inflammation was filled with mice on the third day after infection. The lung inflammation in the TCM group was more reduced than model group, with less inflammatory exudate and only scattered inflammatory exudates. Meanwhile, pulmonary inflammatory pathology was reduced and the amount of bacteria were reduced in the lungs of TCM group (P<0.05), but it was less effective than linezolid (P<0.05). Conclusion Qingjin Granules can significantly reduce the microbial load of the lung and infiltration of inflammatory cells in lung tissue of mice, achieving the efficacy of treating MRSA pneumonia in mice.

Assessment of Height Prediction Model Based on SNPs Loci / 法医学杂志

Objective To establish a height prediction model of Chinese Han male based on the reported 547 height-associated single nucleotide polymorphisms (SNPs) loci in Europeans, and assess its accuracy for height estimation. Methods The DNA typing was analyzed in 59 Han male samples of Shandong province by Affymetrix SNP Array 6.0 chip and HiSeq 4000 sequencing platform. Prediction model was established using 547 height-associated SNPs loci as predictors and weight allele sums (WAS) as com-puting method. The accuracy of height prediction model was analysed using receiver operating characteris-tic (ROC) curve and area under curve (AUC). Results There was no height-associated SNPs locus was found by genome-wide association studies. In present study, height prediction model was established by WAS and obtained an AUC of 0.67 (95％CI:0.53-0.90). Conclusion It has reference value for predicting the height of Han male in Shandong province by WAS model based on 547 SNPs loci, while it is still necessary to further promote the accuracy of the prediction model by screening more height-associated SNPs loci with population heterogeneity.

The self-powered and implantable stimulator driven by infrared ray / 西安交通大学学报(医学版)

Objective To explore the feasibility of fabricating a self-powered and implantable stimulator . Methods Based on pyroelectric effect and high penetrability of infrared ray , an implantable stimulator was designed and fabricated .The electrical performance of stimulator was measured under infrared ray illumination .We conducted the animal experiments in vitro and in vivo to observe the response of gastrocnemius contraction under stimulation .Results The stimulator could output the electrical signal under the periodical infrared ray illumination .The output voltage and current were proportional to the intensity of infrared ray ,which could reach up to 1 .2 V . A real-time electrical stimulation of frog gastrocnemius was conducted and obvious contraction was observed .The tension values increased with the increase of infrared intensity .We further carried out the in vivo experiment with a frog in order to evaluate the performance of the stimulator after being implanted into the body . The gastrocnemius would also be made to contract even though the infrared intensity decayed when through the skin .Conclusion The proposed pyroelectric stimulator can be self-powered and controlled through near-infrared illumination .This study can provide some guidance for solving the problems of implantable power .

Clinic Efficacy and Action Mechanism of Promoting Repairment of Knee Osteoarthritis's Cartilage Corpuscle via Modified Duhuo Jisheng Mixture / 中国中医药信息杂志

Objective To discuss the clinic efficacy and its mechanism of action in promoting the cell proliferation of knee osteoarthritis's cartilage corpuscle via modified Duhuo Jisheng Mixture. Methods Totally 60 cases were divided into treatment group and control group by random number table method, with 30 cases in each group. Treatment group was given modified Duhuo Jisheng Mixture, 62.5 mL each time, twice a day, orally, and sulfoglycan sulfate capsules, 1 capsule a day, three times a day, orally; while control group was given diacerein capsules, 1 pill a day, twice a day, orally, and sulfoglycan sulfate capsules, the same as treatment group. Patients were required to do quadriceps muscle systolic and functional exercise, and keep warm. The treatment lasted for 6 weeks. VAS score and WOMAC of the two group were observed before and after treatment. IL-1, NO, Wnt5a, β-catenin, Sox9, Collagen Ⅱ in knee joint fluid were detected. Adverse reactions were monitored. Results Compared with pre-treatment, the contents of IL-1, NO, Wnt5a and β-catenin in two groups were reduces (P<0.05); while the contents of Sox9 and Collagen Ⅱ were raised (P<0.05). The VAS scores and WOMAC were reduced in two groups after treatment (P<0.05). There was no statistical significance between the two group in the above indexes (P>0.05). There were no obvious adverse reactions in both groups.Conclusion Modified Duhuo Jisheng Mixture can reduce inflammatory factors of the joint fluid and inhibit the Wnt signal pathway through regulating the expression of Sox9 and Collagen Ⅱ in the pathway, which can promote the cell proliferation of knee osteoarthritis's cartilage corpuscle and benefit the repairment of cartilage corpuscle in order to relieve pains and improve functions of knee joint.

Role of kallistatin in protecting hepatic stellate cells against oxidation through inhibition of oxidative stress and activation of Akt-eNOS signaling / 药学学报

The present study was aimed to investigate the role and mechanisms of kallistatin in protection against oxidative stress-induced hepatic stellate cell damage. The effects of kallistatin on the viability, the intracellular superoxide level and Akt, eNOS molecules were investigated in human hepatic stellate cell line LX-2 and the incompletely activated primary rat hepatic stellate cells. Two different oxidative-stress related models, the hydrogen peroxide model and the iron-overload model were used in the experiments. The results show that kallistatin protected the hepatic stellate cells from oxidative damage and repaired the cell damage by oxidative stress. The main mechanism is antioxidant activity of kallistatin, which can remove the oxidized substances inside the cells. On the other way, kallistatin activates Akt and eNOS molecules to generate the antioxidant effect. Our results help to explore new anti-fibrotic targets.

Comparative study of degenerated versus non-degenerated nucleus pulposus cells in inducing mesenchymal stem cells differentiation / 医学研究生学报

Objective Normal nucleus pulposus cells (nNPCs) can induce the differentiation of mesenchymal stem cells (MSCs).However, the inductive effect of degeneration nucleus pul-posus cells ( dNPCs ) on MSCs has not been determined .This study aimed to compare nNPCs with dNPCs in inducing MSCs into nucleus pulposuslike cells. Methods MSCs (cell line) were co-cultured with NPCs (cell line/isolated from patients with intervertebral disc degeneration) in 3D in vitro.The cells were divided into 5 groups:nNPCs control group, dNPCs control group, MSCs control group, nNPCs-MSCs group (MSCs induced by nNPCs), dNPCs-MSCs group (MSCs induced by dNPCs).After co-cultured for 7 days, the expression level of aggrecan (ACAN), type II collagen (COL-2), SOX-9, matrix metalloproteinase-1 (MMP-1) and MMP-3 in all the 5 groups'cellswere detected by RT-PCR and Western-blot. Results The RT-PCR results showed the expression of ACAN , COL-2 and SOX-9 in nNPCs-MSCs group was higher than that in dNPCs control group and dNPCs -MSCs group (all P<0.05).The expression of MMP-1 and MMP-3 in nNPCs-MSCs group and dNPCs-MSCs group was lower than that in dNPCs control group ( all P<0.01) , but there was no difference when compared with nNPCs control group and MSCs control group (all P>0.05).Western-blot analysis showed that the expression of ACAN , COL-2 and SOX-9 in nNPCs-MSCs group was higher than that in dNPCs control group and dNPCs-MSCs group (all P<0.05).The expression of MMP-1 and MMP-3 in nNPCs-MSCs group and dNPCs-MSCs group was lower than that in nNPCs control group and dNPCs control group ( all P<0.01) , but there was no difference compared with MSCs control group ( all P>0.05) . Conclusion Both nNPCs and dNPCs could induce the differentiation of MSCs into nucleus pulposus cells .After co-cultured with nNPCs, the expression level of ACAN, COL-2 and SOX-9 of MSCs was similar to that of nNPCs, which was superior to the induc-tion of MSCs by dNPCs under the same culture conditions .

Prognostic Value of EZH2,Ki-67 and CD34 Expression in Patients with Surgically Treated Colorectal Cancer / 中山大学学报(医学科学版)

[Objective]To evaluate the expression of enhancer of zeste homolog 2(EZH2),Ki-67 and intratumoral microvaseu?lar density(iMVD)in human colorectal cancer(CRC),and discuss their relationship with the biological behavior and prognosis.[Methods]The expression of EZH2,Ki-67 and iMVD(labeled by CD34 protein)was measured with immunohistochemical MaxVision method in CRC patients followed up over 5 years.[Results]Positive expression rates of EZH2,Ki-67 and iMVD were 40.91%(45/110),57.27%(63/110)and 46.36%(51/110),respectively. EZH2 expression was positively correlated with distant metastasis(P=0.024). Ki-67 expression was positively correlated with tumor size,infiltration depth,AJCC stage and differentiation(P=0.033~0.015). The iMVD expression was positively correlated with infiltration depth ,AJCC stage and differentiation (P=0.016~0.034). Spearman correlation analysis showed that EZH2 expression was positively correlated with Ki-67 expression(r=0.195,P=0.041), whereas negatively correlated with iMVD expression. Ki-67 expression had a positive correlation with iMVD(r=0.213,P=0.025). Multivariate analysis suggested that the EZH2 expression was an adverse independent factor for survivals of CRC patients(HR 1.965, 95%CI:1.019-3.789,P=0.044 for OS).[Conclusion]EZH2 expression was significantly correlate with the proliferation of tumor cells and may plays an important role in the metastasis and prognosis of CRC.

Role of up-regulation of THEM4 on collagen secretion in human renal tubular epithelial cells treated with high glucose / 中国药理学通报

Aim To investigate the effect of thioester-ase superfamily member 4(THEM4)expression on col-lagen secretion in human renal proximal tubular epithe-lial cells (HKC)treated with high glucose. Methods In order to examine the direct effect of THEM4 ex-pression vector on PI3K/ Akt pathway and collagen se-cretion,pYr-ads-4-THEM4 expression vector was con-structed and transfected into the HKC with lipo-fectamine 2000 in vitro. HKC cells were randomly di-vided into four groups:normal glucose group (Con-trol),high glucose group (HG),high glucose plus pYr-ads-4-THEM4 vector group (HG + THEM4 vec-tor) and high glucose plus pYr-adshuttle-4 vector group (HG + V vector). After 48 h with HG stimula-tion,the cells were collected for extraction of protein and phospho-Akt (Ser 473),THEM4,TGF-β1 andα-SMA protein expression were examined by Western blot and immunofluorescence staining respectively. Col Ⅰ and Col Ⅲ were detected using the competitive sandwich ELISA kit according to the manufacturer's instructions. Results High glucose inhibited THEM4 expression,and induced increased phospho-Akt (Ser 473),TGF-β1,α-SMA and secreted ColⅠand secre-ted Col Ⅲ in HKC cells. Up-regulation of THEM4 re-versed high glucose-induced decreased THEM4,in-creased phospho-Akt (Ser 473),TGF-β1,α-SMA, secreted Col Ⅰ and secreted Col Ⅲ in HKC cells. Conclusion The up-regulation of THEM4 may de-crease Col Ⅰ and Col Ⅲ secretion by inhibiting the phosphorylation of Akt and down-regulating the expres-sion of TGF-β1 and α-SMA in high glucose-induced HKC cells.