Target versus sub-target dose of renin-angiotensin system inhibitors on survival in elderly patients with heart failure with reduced ejection fraction: a systematic review and meta-analysis / 老年心脏病学杂志（英文版）

BACKGROUND@#The efficiency of the target versus sub-target dose of renin-angiotensin system inhibitors (RASIs) in elderly patients with heart failure (HF) with reduced ejection fraction (HErEF) remains unclear.@*METHODS@#PubMed, Embase, and the Cochrane Central Register of Controlled Trials were searched from database inception through March 2022 for randomized controlled trials (RCTs) and observational studies considering the effect of the target versus sub-target dose of RASIs on survival in elderly patients (≥ 60 years) with HErEF. The primary outcome was all-cause mortality. The secondary outcomes were cardiac mortality, HF hospitalization, and the composite endpoint of mortality or HF hospitalization. A meta-analysis was conducted to generate combined hazard ratio (HR) and 95% CI.@*RESULTS@#Seven studies (two RCTs and five observational studies) enrolling 16,634 patients were included. A pooled analysis suggested that the target versus sub-target dose of RASIs led to lower rates of all-cause mortality (HR = 0.92, 95% CI: 0.87-0.98, I2 = 21%) and cardiac mortality (HR = 0.93, 95% CI: 0.85-1.00, I2 = 15%) but not reduced rates of HF hospitalization (HR = 0.94, 95% CI: 0.88-1.01, I2 = 0) and the composite endpoint (HR = 1.03, 95% CI: 0.91-1.15, I2 = 51%). However, the target dose of RASIs was associated with a similar primary outcome (HR = 0.85, 95% CI: 0.64-1.14, I2 = 0) in a subgroup of very elderly patients > 75 years of age.@*CONCLUSIONS@#Our analysis suggests that the target dose of RASIs has a better survival benefit in elderly patients with HFrEF compared to the sub-target dose of RASIs. However, the sub-target dose of RASIs is associated with a similar mortality rate in very elderly patients > 75 years of age. Future high-quality and adequately powered RCTs are warranted.

Ketogenic diet improves low temperature tolerance in mice by up-regulating PPARα in the liver and brown adipose tissue / 生理学报

The aim of the present study was to investigate the effects of short-term ketogenic diet on the low temperature tolerance of mice and the involvement of peroxisome proliferator-activated receptor α (PPARα). C57BL/6J mice were divided into two groups: normal diet (WT+ND) group and ketogenic diet (WT+KD) group. After being fed with normal or ketogenic diet at room temperature for 2 d, the mice were exposed to 4 °C low temperature for 12 h. The changes in core temperature, blood glucose, blood pressure of mice under low temperature condition were detected, and the protein expression levels of PPARα and mitochondrial uncoupling protein 1 (UCP1) were detected by Western blot. PPARα knockout mice were divided into normal diet (PPARα-/-+ND) group and ketogenic diet (PPARα-/-+KD) group. After being fed with the normal or ketogenic diet at room temperature for 2 d, the mice were exposed to 4 °C low temperature for 12 h. The above indicators were also detected. The results showed that, at room temperature, the protein expression levels of PPARα and UCP1 in liver and brown adipose tissue of WT+KD group were significantly up-regulated, compared with those of WT+ND group. Under low temperature condition, compared with WT+ND, the core temperature and blood glucose of WT+KD group were increased, while mean arterial pressure was decreased; The ketogenic diet up-regulated PPARα protein expression in brown adipose tissue, as well as UCP1 protein expression in liver and brown adipose tissue of WT+KD group. Under low temperature condition, compared to WT+ND group, PPARα-/-+ND group exhibited decreased core temperature and down-regulated PPARα and UCP1 protein expression levels in liver, skeletal muscle, white and brown adipose tissue. Compared to the PPARα-/-+ND group, the PPARα-/-+KD group exhibited decreased core temperature and did not show any difference in the protein expression of UCP1 in liver, skeletal muscle, white and brown adipose tissue. These results suggest that the ketogenic diet promotes UCP1 expression by up-regulating PPARα, thus improving low temperature tolerance of mice. Therefore, short-term ketogenic diet can be used as a potential intervention to improve the low temperature tolerance.

Sacubitril/valsartan attenuates left ventricular remodeling and improve cardiac function by upregulating apelin/APJ pathway in rats with heart failure / 中华心血管病杂志

Objective: To investigate the effect and mechanism of sacubitril/valsartan on left ventricular remodeling and cardiac function in rats with heart failure. Methods: A total of 46 SPF-grade male Wistar rats weighed 300-350 g were acclimatized to the laboratory for 7 days. Rats were then divided into 4 groups: the heart failure group (n=12, intraperitoneal injection of adriamycin hydrochloride 2.5 mg/kg once a week for 6 consecutive weeks, establishing a model of heart failure); heart failure+sacubitril/valsartan group (treatment group, n=12, intragastric administration with sacubitril/valsartan 1 week before the first injection of adriamycin, at a dose of 60 mg·kg-1·d-1 for 7 weeks); heart failure+sacubitril/valsartan+APJ antagonist F13A group (F13A group, n=12, adriamycin and sacubitril/valsartan, intraperitoneal injection of 100 μg·kg-1·d-1 APJ antagonist F13A for 7 weeks) and control group (n=10, intraperitoneal injection of equal volume of normal saline). One week after the last injection of adriamycin or saline, transthoracic echocardiography was performed to detect the cardiac structure and function, and then the rats were executed, blood and left ventricular specimens were obtained for further analysis. Hematoxylin-eosin staining and Masson trichrome staining were performed to analyze the left ventricular pathological change and myocardial fibrosis. TUNEL staining was performed to detect cardiomyocyte apoptosis. mRNA expression of left ventricular myocardial apelin and APJ was detected by RT-qRCR. ELISA was performed to detect plasma apelin-12 concentration. The protein expression of left ventricular myocardial apelin and APJ was detected by Western blot. Results: Seven rats survived in the heart failure group, 10 in the treatment group, and 8 in the F13A group. Echocardiography showed that the left ventricular end-diastolic diameter (LVEDD) and the left ventricular end-systolic diameter (LVESD) were higher (both P<0.05), while the left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were lower in the heart failure group than in the control group (both P<0.05). Compared with the heart failure group, rats in the treatment group were featured with lower LVEDD and LVESD (both P<0.05), higher LVEF and LVFS (both P<0.05), these beneficial effects were reversed in rats assigned to F13A group (all P<0.05 vs. treatment group). The results of HE staining showed that the cardiomyocytes of rats in the control group were arranged neatly and densely structured, the cardiomyocytes in the heart failure group were arranged in disorder, distorted and the gap between cells was increased, the cardiomyocytes in the treatment group were slightly neat and dense, and cardiomyocytes in the F13A group were featured similarly as the heart failure group. Masson staining showed that there were small amount of collagen fibers in the left ventricular myocardial interstitium of the control group, while left ventricular myocardial fibrosis was significantly increased, and collagen volume fraction (CVF) was significantly higher in the heart failure group than that of the control group (P<0.05). Compared with the heart failure group, the left ventricular myocardial fibrosis and the CVF were reduced in the treatment group (both P<0.05), these effects were reversed in the F13A group (all P<0.05 vs. treatment group). TUNEL staining showed that the apoptosis index (AI) of cardiomyocytes in rats was higher in the heart failure group compared with the control group (P<0.05), which was reduced in the treatment group (P<0.05 vs. heart failure group), this effect again was reversed in the F13A group (P<0.05 vs. treatment group). The results of RT-qPCR and Western blot showed that the mRNA and protein levels of apelin and APJ in left ventricular myocardial tissue of rats were downregulated in heart failure group (all P<0.05) compared with the control group. Compared with the heart failure group, the mRNA and protein levels of apelin and APJ were upregulated in the treatment group (all P<0.05), these effects were reversed in the F13A group (all P<0.05 vs. treatment group). ELISA test showed that the plasma apelin concentration of rats was lower in the heart failure group compared with the control group (P<0.05); compared with the heart failure group, the plasma apelin concentration of rats was higher in the treatment group (P<0.05), this effect was reversed in the F13A group (P<0.05 vs. treatment group). Conclusion: Sacubitril/valsartan can partially reverse left ventricular remodeling and improve cardiac function in rats with heart failure through modulating Apelin/APJ pathways.

Exercise preconditioning attenuates the lipid metabolic disorder induced by chronic high fat diet feeding in mice / 生理学报

Mounting evidence has shown that exercise exerts extensive beneficial effects, including preventing and protecting against chronic diseases, through improving metabolism and other mechanisms. Recent studies have shown that exercise preconditioning affords significant cardioprotective effects. However, whether exercise preconditioning improves high fat diet (HFD)-induced obesity and lipid metabolic disorder remains unknown. The study was aimed to explore the effects of exercise preconditioning on HFD-induced obesity and lipid metabolic disorder in mice. 4-week-old C57BL/6 mice were subjected to swimming or sedentary control for 3 months, and then were fed with normal diet (ND) or HFD for 4 more months. The results showed that the blood glucose was decreased, and the glucose tolerance and grip strength were increased in exercised mice after training. Exercise preconditioning failed to improve HFD-induced body weight gain, but improved HFD-induced glucose intolerance. Exercise preconditioning showed no significant effects on both exercise capacity and physical activity in ND- and HFD-fed mice. HFD feeding increased total cholesterol and low density lipoprotein (LDL) levels in circulation, promoted subcutaneous fat and epididymal fat accumulation in mice. Exercise preconditioning increased circulating high density lipoprotein (HDL) and decreased circulating LDL, without affecting the subcutaneous fat and epididymal fat in HFD-fed mice. HFD feeding increased liver weight and hepatic total cholesterol contents, and dysregulated the expressions of several mitochondria function-related proteins in mice. These abnormalities were partially reversed by exercise preconditioning. Together, these results suggest that exercise preconditioning can partially reverse the HFD-induced lipid metabolic disorder and hepatic dysfunction, and these beneficial effects of exercise sustain for a period of time, even after exercise is discontinued.

Ecological Suitability Regionalization of Ziziphus jujuba / 中国实验方剂学杂志

Objective:To analyze the main factors affecting the <italic>Ziziphus jujuba</italic> distribution and expand the understanding of its distribution and the corresponding influencing factors by comparing the distribution sites of <italic>Z. jujuba</italic> predicted by models with those recorded in the literature. Method:More than 200 distribution sites of <italic>Z. jujuba</italic> accompanied by 55 environmental factors were obtained from literature and specimen review. The environmental factors that affect the distribution of <italic>Z. jujuba</italic> were explored by maximum entropy （MaxEnt） model， and the potential distribution areas of <italic>Z. jujuba</italic> in China were analyzed by ArcGIS， followed by the verification of the main environmental factors using receiver-operating characteristic （ROC） curve and Jackknife method. Result:The area under the curve （AUC） values for the test data and training data were both greater than 0.9， which perfectly satisfied the standard， indicating that the research results were accurate and reliable. Conclusion:The annual average temperature， the average temperature in May， the average temperature in the warmest season， vegetation type， soil type， average temperature in June， average temperature in September， and average temperature in August are proved to be the main environmental factors affecting the distribution of <italic>Z. jujuba</italic>， which can be found almost all over China， except for Heilongjiang and Tibet. <italic>Z. jujuba</italic> is most suitable to be planted in southeastern Sichuan， Chongqing， southern Gansu， Ningxia， most areas of central Shaanxi， eastern and southwestern Shanxi， Henan， eastern and northern Hubei， northern and eastern Anhui， Shandong， Hebei， Beijing， Tianjin， western Liaoning， and Zhejiang. As revealed by literature review， the most suitable growing areas of <italic>Z. jujuba</italic> are southeastern Sichuan， central Shaanxi， southwestern Shanxi， western and northern Henan， Shandong， and southwestern and eastern Hebei.

Silybin alleviates hepatic lipid accumulation in methionine-choline deficient diet-induced nonalcoholic fatty liver disease in mice via peroxisome proliferator-activated receptor α / 中国天然药物

Nonalcoholic fatty liver disease (NAFLD) is regarded as the most common liver disease with no approved therapeutic drug currently. Silymarin, an extract from the seeds of Silybum marianum, has been used for centuries for the treatment of various liver diseases. Although the hepatoprotective effect of silybin against NAFLD is widely accepted, the underlying mechanism and therapeutic target remain unclear. In this study, NAFLD mice caused by methionine-choline deficient (MCD) diet were orally administrated with silybin to explore the possible mechanism and target. To clarify the contribution of peroxisome proliferator-activated receptor α (PPARα), PPARα antagonist GW6471 was co-administrated with silybin to NAFLD mice. Since silybin was proven as a PPARα partial agonist, the combined effect of silybin with PPARα agonist, fenofibrate, was then evaluated in NAFLD mice. Serum and liver samples were collected to analyze the pharmacological efficacy and expression of PPARα and its targets. As expected, silybin significantly protected mice from MCD-induced NAFLD. Furthermore, silybin reduced lipid accumulation via activating PPARα, inducing the expression of liver cytosolic fatty acid-binding protein, carnitine palmitoyltransferase (Cpt)-1a, Cpt-2, medium chain acyl-CoA dehydrogenase and stearoyl-CoA desaturase-1, and suppressing fatty acid synthase and acetyl-CoA carboxylase α. GW6471 abolished the effect of silybin on PPARα signal and hepatoprotective effect against NAFLD. Moreover, as a partial agonist for PPARα, silybin impaired the powerful lipid-lowering effect of fenofibrate when used together. Taken together, silybin protected mice against NAFLD via activating PPARα to diminish lipid accumulation and it is not suggested to simultaneously take silybin and classical PPARα agonists for NAFLD therapy.

Ecology suitability study of Polygonatum cyrtonema / 中国中药杂志

Using the 260 geographical distribution records of Polygonatum cyrtonema in China, combined with 53 environmental factors, the maximum entropy modeling(MaxEnt) was used to study the ecological factors affecting the suitability distribution of P. cyrtonema. The ArcGIS software was used to predict the potential distribution of the population of P. cyrtonema. The dominant factors were chosen by using the Jackknife test and the Receiver Operating Characteristic(ROC) curve was used to evaluate the simulation. The results showed that high value of area under curve(AUC) denoted good results, which significantly differed from random predictions. Based on the evaluation criterion, the accuracies of the predictions of P. cyrtonema potential distribution in the current periods were excellent. The main environmental factors affecting the suitable growth of P. cyrtonema were the monthly precipitation, the wettest monthly precipitation, the annual average temperature range and the precipitation of November, March, February, April, May and October. There are 9 environmental factors in soil type. The potential fitness of P. cyrtonema in China is high, mainly concentra-ted in Hunan, western Hubei, Guangdong, northeastern Guangxi, southeastern Guizhou, Jiangxi, southwestern Anhui, Fujian, Zhejiang, Shaanxi, southwestern Henan and Chongqing. The growth distribution of the potential distribution area of P. cyrtonema was divided, and the zoning map of the growth suitability of P. cyrtonema was formed. Through the comparative analysis of the potential distribution range based on MaxEnt and the distribution range of literature records, the understanding of the distribution range of P. cyrtonema was expanded.

Identification of Astragalus and its adulterants based on ITS2sequence and secondary structure / 药学学报

To effectively identify the Astragalus and its adulterants based on ITS2 sequence and secondary structure, in this study, 32 portions of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Beg.) Hsiao and Astragalus membranaceus (Fisch.) Bge. collected were conducted ITS2 sequence amplification and bidirectional sequencing, whose results were then spliced by CExpress software remove the 5.8S and 28S sequences at both ends to obtain a complete ITS2 sequence. In addition, 3 ITS2 sequences for each of the adulterants of Astragalus, respectively, Oxytropis coerulea, Caragana sinica, Hedysarum polybotrys, Althaea rosea were downloaded from GenBank. The intra-specific and inter-specific genetic distances were calculated by the software MEGA7 to analyze the difference of each sequence; the Neighbor-joining (NJ) method was used to construct the phylogenetic tree based on ITS2 sequence (primary structure) as well as joint ITS2 sequence and its secondary structure. The results showed that the average ITS2 sequence length of both A. mongolicus and A. membranaceus was 216 bp, and their average GC content was 50.00% and 50.46%, respectively. The similarity of ITS2 sequence length and GC content between the two kind of Astragalus and Oxytropis coerulea was the highest, while the ITS2 sequence length and GC content of Althaea rosea showed great differences with those of Astragalus. The inter-specific distance between Astragalus and Oxytropis coerulea was the smallest, while that between the medicinal Astragalus and Hedysarum polybotrys, Caragana sinica as well as Althaea rosea was great. The phylogenetic trees constructed based on the ITS2 sequence (primary structure) and joint ITS2 sequence and its secondary structure showed that the topological relations of the two phylogenetic trees were basically the same, and both could effectively identify the Astragalus and its adulterants. What’s more, the addition of secondary structure information made end branch of the phylogenetic tree become more in its construction, and the distinguish ability and approval rating were also improved, which further reflected the genetic relationship of Astragalus and its adulterants. This provides some scientific basis for classification and accurate identification of Astragalus and its adulterants.

Reseach on origin identification of commercially available Zaocys dhumnades / 中国中药杂志

Through market investigation, the adulteration of Zaocys dhumnades on markets was found out, and samples of authentic and adulterated Z. dhumnades on markets were collected. The origin and properties of the adulterated Z. dhumnades were studied in order to provide reference for the identification of Z. dhumnades. The counterfeit Z. dhumnades sold on markets were as follows: Ptyas korros, P. mucosus, Najanaja atra, Sinonatrix annularis, Dinodon septentrionalis, etc. It is found that there existed a obvious difference between the traits of the Z. dhumnades and counterfeits. Genuine Z. dhumnades with "sword ridge" "iron tail", strongly ribbed scales and other features, is the key point to identify the difference from adulterants.

Identification and antimicrobial resistance of Escherichia coli isolates from chicken in different provinces / 中国人兽共患病学报

The aim of this study was to determine the prevalence and antibiotic susceptibility of Escherichia coli from retail raw chickens in different provinces of China.A total of 1 152 whole chicken samples were collected and screened for the prevalence of E.coli,and then the E.coli isolates were further tested for the antimicrobial susceptibility using agar dilution method.Results showed that the overall positive rate for E.coli in retail chickens was 65.97％ (760/1 152);resistance was most common to nalidixic acid (66.84％),followed by amoxicillin-clavulanic acid (66.05％),tetracycline (65.00％),trimethoprim-sulfamethoxazole (63.16 ％),ampicillin (60.66 ％),amoxicillin (51.32 ％),streptomycin (50.39 ％),chloramphenicol (48.32 ％),kanamycin (38.29 ％),gentamicin (26.31％),ciprofloxacin (25.79 ％) and cefoxitin (21.05 ％).The last were gatifloxacin,cefoperazone and amikacin (each ＜20％).Overall,70.53％ of the isolates were resistant to at least three antimicrobials.A large proportion of multidrug resistant isolates were resistant to 8 kinds of antimicrobials (10.26 ％).No strain was resistant to 15 kinds of antimicrobials.Furthermore,isolates recovered from different regions exhibited different resistance levels to most antimicrobials.Our findings indicate that retail chicken in China was commonly contaminated with E.coli,and many E.coli strains exhibited multiple drug resistance.Presence of multiple drug E.coli in raw chickens may pose a potential threat to human health.

Cumulative Meta-analysis on Clinical Efficacy of Relieving Fever with Sweet and Warm Drugs for Noninfectious Fever Caused by Vital-energy Deficiency / 中国中医药信息杂志

Objective To systematically evaluate the clinical efficacy of relieving fever with sweet and warm drugs (Ganwen Chure Therapy) for noninfectious fever caused by vital-energy deficiency based on Meta-analysis. Methods Literature about RCTs of Ganwen Chure Therapy for noninfectious fever caused by vital-energy deficiency in CNKI, Wanfang Database, CBM, VIP, Pub Med, Cochrane Library, and Embase was retrieved by computers from establishment of database to April 2017. After two researchers independently conducted literature screening, cross-checking, data extraction, and literature quality evaluation, cumulative Meta-analysis was performed on the outcome indicators in order of publication time and sample size, and the trend of the results was tested. Then the quality of the literature based on GRADE was under an overall evaluation. Results Totally 27 articles were included in this study, involving 2599 patients. The results of cumulative Meta-analysis showed that the total effective rates of using Ganwen Chure Therapy only [OR=3.875, 95％CI (2.87, 5.24), Z=8.82, P=0.000 1]and Ganwen Chure Therapy combined with routine therapy [OR=5.791, 95％CI (3.55, 9.45), Z=7.03, P=0.000 1]were better than the routine therapy, with statistical significance. Trend test showed that there was variability in the timing of drug combination study, showing that the cumulative Meta-analysis results were not stable. Conclusion Ganwen Chure Therapy has certain efficacy for noninfectious fever caused by vital-energy deficiency. However, the overall quality of the included studies was low, with relatively high homogeneity. There are biases in publication, yet more high-quality clinical research is needed for further verification.

Effects of endostatin on C6 glioma-induced edema / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Glioma-induced edema is considered as one of the most pathological characteristics of glioma and a significant source of morbidity and mortality. New strategies are needed for the treatment of peritumoral edema in glioma. Endostatin has been proven to be beneficial as an anti-angiogenic agent in experimental gliomas, but the effects are unclear. This study aimed to investigate the effects of endostatin on C6 glioma-induced edema.</p><p><b>METHODS</b>Tumorigenic mice were established by subcutaneous injection of three glioma cell lines, C6-null cells and stable transfected-C6 cells overexpressing mock vector (C6-mock cells) and endostatin (C6-endo cells). Endostatin expression in xenograft C6 glioma was determined by immunostaining and Western blotting. Glioma-induced edema and tumor vessel permeability were assayed. The effect of endostatin on vascular enodothelial growth factor (VEGF) expression in vivo was analyzed by quantitative polymerase chain reaction (Q-PCR) and enzyme-linked immunosorbent assay (ELISA). The number of vesiculo-vascuolar organelles (VVOs) formed in tumor endothelia was calculated using electron microscopy. Data were analyzed by using one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test for multiple comparisons to the control groups.</p><p><b>RESULTS</b>Overexpression of endostatin (C6-endo cells) significantly suppressed tumor growth and reduced tumor edema and vessel permeability. ELISA analysis showed that the level of VEGF protein was markedly decreased in tumor from C6-endo cells compared with tumor from C6-null cells and C6-mock cells. Similar results were obtained by Q-PCR. Furthermore, the number of VVOs observed in tumor from C6-endo mice was significantly reduced compared with tumor from C6-null cells or C6-mock cells.</p><p><b>CONCLUSIONS</b>Our data provide primary evidence that endostatin reduces glioma-induced edema and vascular permeability. Using endostatin may be an effective strategy for treating glioma edema.</p>

Screening for differentially expressed proteins in nasopharyngeal carcinoma by laser capture microdissection and proteomic analysis / 中南大学学报(医学版)

OBJECTIVE@#To search for the differentially expressed proteins of nasopharyngeal carcinoma (NPC),and provide scientific evidence for identifying molecular biomarkers for NPC.@*METHODS@#Laser capture microdissection (LCM) was used to purify the target cells from NPC and normal nasopharyngeal epithelial tissues (NNET). Two-dimensional gel electrophoresis (2-DE) was used to separate the total proteins of microdissected NPC and NNET, PDQuest software was applied to analyze 2-DE images,and the differential proteins between the 2 types of tissues were identified by both MALDI-TOF-MS and ESI-Q-TOF-MS. Western blot and immunohistochemistry of tissue microarray were used to detect the expression of the differential protein SCCA1 in NPC and NNET.@*RESULTS@#2-DE patterns of microdissected NPC and NNEC were established,and 36 differential proteins in the NPC and NNEC were identified,20 of which only expressed or up-regulated in NPC and 16 only expressed or up-regulated in NNET. The differentially expressed level of SCCA1 in the NPC and NNET was confirmed by Western blot and immunohistochemistry of tissue microarray.@*CONCLUSION@#Thirty-six differentially expressed proteins identified in this study may be associated with the carcinogenesis of NPC,and may be candidate molecular biomarkers for NPC.

Differential proteomic analysis of nasopharyngeal carcinoma cell lines 5-8F and 6-10B / 中南大学学报(医学版)

OBJECTIVE@#To compare the proteome difference of nasopharyngeal carcinoma (NPC) cell lines 5-8F and 6-10B, and to screen these proteins associated with NPC metastasis.@*METHODS@#Two-dimensional gel electrophoresis (2-DE) was used to separate the total proteins from NPC cell lines 5-8F and 6-10B with different metastatic potentials and same genetic background, respectively. PDQuest software was applied to analyze 2-DE images, and the differentially expressed protein spots between 5-8F and 6-10B were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The expression levels of partial identified proteins in the 2 cell lines were detected by Western blot.@*RESULTS@#2-DE maps of total proteins from 5-8F and 6-10B were established. A total of 65 differential protein spots in the 2 cell lines were found, and 15 non-redundant differential expression proteins were identified by MALDI-TOF-MS. Western blot showed that Annexin A1 and 14-3-3 protein sigma were differential expression proteins in 5-8F and 6-10B, which was consistent with the Results from the comparative proteomic analysis.@*CONCLUSION@#Fifteen non-redundant differential expression proteins are useful for studying the metastatic mechanism of NPC.

Mechanism of hepatocyte transformation by HCV NS3 using two-dimensional electrophoresis and mass spectrometry / 中南大学学报(医学版)

OBJECTIVE@#To investigate the proteome of hepatocyte transformation by hepatitis C virus (HCV) nonstructural protein 3 (NS3).@*METHODS@#Human hepatocyte line QSG7701 stably expressing HCV NS3 C-terminal deleted protein was constructed, which was named pRcHCNS3/QSG. Two-dimensional electrophoresis (2-DE) was used to separate the total protein of pRcHCNS3/QSG and pRcCMV transfected cells (pRcCMV/QSG) respectively. Differentially expressed protein spots were identified by mass spectrometry. Western blot confirmed the differentially expressed proteins.@*RESULTS@#2-DE profiles with high resolution and reproducibility were obtained. The average spots of pRcHCNS3/QSG and pRcCMV/QSG were (1183+/-77) and (1095+/-82) respectively, and (920+/-60) spots were matched. Twenty-one differentially expressed protein spots were chosen randomly and 15 were identified by mass spectrometry. Some proteins such as Ras, P38 and HD53 which were involved in signal transduction were increased in pRcHCNS3/QSG cells. Western blot also showed strong expression of phosphorylated P44/42 and P38 in pRcHCNS3/QSG cells. Other differentially expressed proteins were related to cell cycle regulation, immunoreaction, tumor invasion and metastasis, and liver metabolizability.@*CONCLUSION@#HCV NS3 might be involved in cell malignant transformation through affecting protein expression and signal transduction such as MAPK cascade. Further study on the signal transductions and their relationship would not only be helpful to explore the mechanism of HCV related HCC, but also provide a new idea for the molecular treatment of HCC.

Effects of bone marrow stromal cells on endothelial cells proliferation and microvessel formation in vitro / 中国康复理论与实践

@#ObjectiveTo observe the effects of bone marrow stromal cells (BMSCs) on vessel endothelial cells proliferation and microvessel formation in vitro.MethodsBMSCs and brain vessel endothelial cells were separated from adult and divided into co-culture group of BMSCs and endothelial cells, medium group of BMSCs, comparison group. Endothelial cells proliferation and microvessel formation were observed. ResultsEndothelial cells were promoted to proliferate and formate the microvessel in medium group and co-culture group. And the effect was prominence in co-culture group.ConclusionBMSCs can promote the proliferation and microvessel formation of endothelial cells.

Establishment of protein expression profile of human normal colonic epithelia / 中南大学学报(医学版)

OBJECTIVE@#To establish a protein expression profile of human normal colonic epithelia.@*METHODS@#Two-dimensional gel electrophoresis (2-DE) was applied to separate the total proteins of 20 human normal colonic epithelial tissues. The expression proteins in the human normal colonic epithelia were identified by both matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and electrospray ionization tandem mass spectrometry (ESI-Q-TOF), and the biological function and subcellular locations of the identified proteins were analyzed by bioinformatics.@*RESULTS@#A 2-DE reference map of human normal colonic epithelium was established. On the 2-DE map, 1020+/-50 protein spots were detected, 204 protein spots representing 162 non-redundant proteins were identified, and 37 proteins had posttranslational modification. The identified proteins were categorized into several protein groups according to their functions or subcellular locations, whose data were available at our website (http://www.xyproteomics.org).@*CONCLUSION@#A protein expression profile of human normal colonic epithelia is established for the first time, which provides useful information for investigating the physiological functions and pathologic process of colonic epithelia.

Identification of aging related proteins in human normal colonic epithelium / 中南大学学报(医学版)

OBJECTIVE@#To explore the molecular mechanisms of colonic epithelial aging related proteins and aged colonic epithelial susceptibility to tumor.@*METHODS@#The proteins of normal human colonic epithelial tissue from young and old people were separated by 2-dimensional gel electrophoresis (2DGE), respectively. Then gels were stained by silver, scanned by imagescanner and analyzed with PDQuest software. The differentially expressed protein spots of colonic epithelium between the old and the young groups were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching.@*RESULTS@#Well-resolved and reproducible 2DGE maps of normal human colonic epithelium from the young and the old were acquired. Nineteen more than 2 fold differentially expressed protein spots were identified representing 17 different proteins by MALDI-TOF-MS. The functions of these proteins involve in metabolism, energy generation, transportation, antioxidation, translation and protein folding.@*CONCLUSION@#Seventeen aging related proteins of human colonic epithelium identified indicate that injury of mitochondrial function and decline of antioxidant capability are important reasons for the aging of human colonic epithelium. These data provided useful clues for elucidating the mechanisms of colonic epithelial aging and aged colonic epithelial susceptibility to cancer.

Expression and purification of varicella-zoster virus glycoprotein I gene in insect cells / 中华预防医学杂志

<p><b>OBJECTIVE</b>To express the cloned gene glycoprotein I (gpI) of varicella-zoster virus (VZV), Beijing VZV 84-7 strain in insect cells and to purify its expression product.</p><p><b>METHODS</b>The gene coding for gpI of VZV was amplified from viral DNA by PCR and cloned into baculovirus transfer vector (pBacPAK9), and recombinant transfer vector plasmid pBacVZVgpI was obtained. The inserted gpI gene in the pBacVZVgpI was sequenced. Insect cells Sf 9 were co-transfected with the recombinant transfer vector plasmid pBacVZVgpI and wild type linear baculovirus BacPAK6 (digested with Bsu36I) DNA. The recombinant baculoviruses containing the VZV 84-7 gpI gene was isolated through several rounds of limited dilution. Recombinant protein gpI was expressed in insect cells Sf 9, postinfected with recombinant baculoviruses. The expressed recombinant gpI was purified by lectin affinity chromatography and its antigenicity and immunogenicity were investigated.</p><p><b>RESULTS</b>The gene coding for gpI of VZV was obtained by PCR and the gpI gene of pBacPAK9 was confirmed by DNA sequencing. The recombinant gpI was expressed in insect cells Sf 9, post-infected with recombinant baculovirus and identified by SDS-PAGE and western blotting, with its product in cell culture reaching the peak in 72 hours and with a molecular mass of 58 kd and 70 kd, the same as theoretical values. Results of immunoassay with cell lysates infected by recombinant baculoviruses indicated that recombinant protein expressed in insect cells had ability of eliciting specific antibodies against native VZV in mice and complement-dependent neutralizing antibodies. The purified recombinant gpI gave a product with a purity of more than 80%. ELISA and Western-blot analysis demonstrated that purified protein had specific VZV antibody-binding activity. This suggested that the recombinant gpI expressed in insect cells had the same biological characteristics as its native counterpart.</p><p><b>CONCLUSION</b>Baculovirus-insect cells could be used to express the gene of VZV gpI, which could provide a basis for quantitative analysis of VZV antigen, and preparation of its subunit vaccine.</p>

Expression of apoptosis and Bcl-2, Bax in hemangioma and vascular malformations / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the expression of apoptosis and their regulatory genes, Bcl-2, Bax in hemangioma and vascular malformations.</p><p><b>METHODS</b>The specimens were taken from 68 cases of strawberry hemangioma or vascular malformations and 11 cases of normal skin. The expression of Bcl-2, Bax and Ki-67 in endothelial cells were investigated by immunohistochemical S-P method. The apoptosis index (AI) in endothelial cells was investigated by TUNEL method.</p><p><b>RESULTS</b>Ki-67 was positive in all the proliferating strawberry hemangioma, negative in the other groups. The expression of Bax was significantly higher in strawberry hemangioma than in vascular malformations and normal skin (P < 0.01). Bcl-2 was negative in all groups. The AI in endothelial cells of strawberry hemangioma was significantly higher than that of vascular malformations and normal skin. There were no statistically significant differences between the proliferating and involution strawberry hemangioma. There were no statistically significant differences between various types of vascular malformations and normal skin. With the increasing of the expression of Bax in strawberry hemangioma, the AI increased.</p><p><b>CONCLUSION</b>These findings clearly demonstrate a much higher apoptotic activity in strawberry hemangioma than in vascular malformations and normal skin and suggest that apoptosis might be a cause of spontaneous involution of strawberry hemangioma and might not be a cause of the pathogenesis of vascular malformation. Bcl-2, Bax might play an important regulatory role in apoptosis in endothelial cells of strawberry hemangioma.</p>