Objective To study the relationship of TNF-? G308A and G238A polymorphism with type 2 diabetes mellitus in elderly patients.Methods Blood specimens were collected from 60 elderly patients with type 2 diabetes mellitus and 30 normal elderly volunteers.Genomic DNA was extracted.The primers of PCR for the SNPs of G308A and G238A were optimized and the polymorphism of amplicons was analyzed by DNA sequencing.Results There was a significant difference of TNF-?-308 gene in DM patients compared with control group,while no difference of TNF-?-238.Conclusions The 308 G-A polymorphism of TNF-? promoter gene may be associated with type 2 diabetes mellitus in elderly patients.

Objective To investigate the effects of rosiglitazone (RSG) on the levels of serum HbA1c, interleukin-6(IL-6) and high sensitive C-reactive protein (hsCRP) in type 2 diabetic patients who were inadequately controlled only with insulin therapy. Methods 40 patients with type 2 diabetes mellitus (T2DM), whose blood glucose was poorly controlled (HbA1c≥7 5%) after 4 weeks of standardized insulin therapy, were additionally given 4mg daily PSG. The levels of serum HbA1c, IL-6, hsCRP, total cholesterol (Tch), triglyceride (TG), low-density lipoprotein cholesterol (LDL-c) and high-density lipoprotein cholesterol (HDL-c) were measured before and after 12 weeks treatment with RSG. hsCRP was measured by high-sensitivity immunoturbidimetric method, and IL-6 by enzyme-linked immunoadsorbent assay (ELISA). Results ⑴ RSG 4mg daily significantly improved glycemic control, treatment with RSG 4 mg plus insulin resulted in a mean 1 59% reduction of HbA1c from baseline (P

Objective To investigate effects and its mechanisms of hypertonic saline hydroxyethyl starch 200/0.5 solution on intracranial pressure and brain water content in rats with ischemic cerebral edema.Methods All experiments were conducted in the animal experimental center of Sun Yat-sen University.The 28 male Sprague-Dawle (SD) rats were randomly (random number) divided into hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group,control group and sham operation group,each n =7.Ischemic cerebral edema model was reproduced by middle cerebral artery occlusion (MCAO),followed by reperfusion after ischemia for 2 hours (If the moldel was not successful,other rats were operated to fill the missing models).Then reperfusion after ischemia 2 hours and received hypertonic saline hydroxyethyl starch and hydroxyethyl starch via tail vein at the beginning of reperfusion.The colloidal osmotic pressure (COP) and intracranial pressure (ICP) were evaluated on 0,2,6,12,18,24 hours after the surgery.The water content of the right hemisphere was measured on 24 h after the surgery.Results The ICP of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 2,6,12,18,24 h after the surgery.The ICP of hypertonic saline hydroxyethyl starch group was significantly lower than those of hydroxyethyl starch group and control group on 2,6,12,18 and 24 h.But there was no significant difference in ICP of the hydroxyethyl starch group compared with that of control group at all time points.The COP of hypertonic saline hydroxyethyl starch group and hydroxyethyl starch group were significantly higher than the control group and sham operation group at each time point; There was no significant difference in COP (mmHg) of the hydroxyethyl starch group compared with that of hypertonic saline hydroxyethyl starch group at all time points.The brain water content (BWC) of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 24 hours after the surgery [(81.24±0.36)％,(83.04±0.10)％,(83.14±0.41)％ vs.(78.37±0.37)％,all P=0.000],BWC of hypertonic saline hydroxyethyl starch group lower than these of hydroxyethyl starch group [(81.24±0.36)％ vs.(83.04 ±0.10) ％,P =0.000] and control group [(81.24 ±0.36)％ vs.(83.14 ±0.41) ％,P =0.000].There was no significant difference in BWC of the hydroxyethyl starch group compared with that of control group [(83.04 ± 0.10) ％ vs.(83.14 ± 0.41) ％,P =0.578].Conclusion Hypertonic saline hydroxyethyl starch solution could significantly ameliorate ischemic cerebral edema and reduce ICP,but the relationship between its elevated COP and reduced ICP has not been confirmed.

Objective To evaluate the relationship of oncosis and the expression of microvessel density (MVD) in colorectal carcinoma and its clinical significance Methods The expression of MVD and oncosis were detected by immunohistochemical method and transmission electron microscope in 64 cases of colorectal carcinoma Results Oncosis existed in colorectal carcinoma Oncosis index (OI) in tissues of colorectal carcinoma decreased with the decrease of differentiation grades ( F=8 590?2, P

Objective To assess the efficacy and safety of urinary kallidinogenase combined with sodium ozagrel for cerebral infarction (CI), and provide references for clinical rational drug use. Methods Retrieved from Cochrane library, PubMed, CBM, FMJS, VIP, Wangfang database and CNKI ( published until January 2015), randomized controlled trails (RCT)about urinary kallidinogenase combined with sodium ozagrel for treatment of CI were included,then methodological quality were evaluated and statistical analysis of those studies were carried out by Rev Man 5.3.4 software. Results 19 RCTs were included,involving 1 747 patients. Results of Meta-analysis showed that urinary kallidinogenase combined with sodium ozagrel could significantly improve total effective rate[RR= 1.18, 95%CI(1.13, 1.23), Z= 7.97, P<0.000 01], cure rate[RR = 1.42, 95%CI(1.23, 1.64), Z= 4.86, P<0.000 1], neurological deficit scores[MD= -4.40, 95%CI(-5.36, -3.43), Z= 8.90,P<0. 000 01] and activity of daily living scores[MD = 19.14, 95%CI(17.39, 20.90), Z = 21.36, P<0.000 01]. Conclusion Urinary kallidinogenase combined with sodium ozagrel was effective in the treatment of CI, and no significant adverse reactions were observed. The combination therapy was worthy of clinical application.

Objective To determine the likelihood of G-protein coupled receptor 56 (GPR56 ) induces axonal development and myelination in the corpus callosum of mouse brain.Methods A total of 64 Gpr56 +/-and Gpr56 -/-mice were selected and randomly divided into two groups:Gpr56 +/-group (n=32)and Gpr56 -/-group (n=32).According to number of days after birth,each group was further divided into 4 subgroups including P7d,P14d,P21d and P28d subgroups.Levels of neurofilament-200 (NF -200)and proteolipid protein (PLP ) of myelin basic protein in corpus callosum were measured with immunohistochemistry staining and Western blot in P7d、P14d、P21d、P28d Gpr56 +/- and Gpr56 -/-mice.Gpr56 +/-and Gpr56 -/-neurons were cultured using P1 d Gpr56 +/-and Gpr56 -/-mouse brain.The lengths of Gpr56 +/- and Gpr56 -/-neuronal axon were measured and compared with Image J software. Axonal myelination in the corpus callosum of mouse brain in each group was observed under electronic microscopy and the axonal diameters between subgroups were compared.Results The levels of NF-200 and PLP in the corpus callosum in P7d、P14d、P21d、P28d Gpr56 -/-mice decreased significantly compared with Gpr56 +/- mice.The length of Gpr56 -/-neuronal axon was shortened compared with Gpr56 +/-neuronal axon.The number of myelinated axons was obviously reduced in the corpus callosum in P28d Gpr56 -/-mice.The diameter of axon in the corpus callosum of P28d Gpr56 +/-mouse is longer than that of P28d Gpr56 -/-mouse. Conclusions GPR56 may be involved in axonal development and myelination in the corpus callosum of mouse brain.

Objective To investigate the clinical value of the ratio of plasma vascular endothelial growth factor level to platelet count (VEGF/PLT) in predicting 28-day prognosis in patients with sepsis.Methods A prospective cohort study was conducted.From September 2009 to March 2013,164 sepsis patients in Intensive Care Unit (ICU) of Guangdong General Hospital were included for study.Patients with age younger than 18 years old,the illness already reaching final stage of chronic diseases,suffering from two or more organs dysfunction within 3 days,acute pancreatitis without infection,or less than 28 days of expected survival time were excluded.Finally,135 patients were included in the further analysis.Peripheral blood samples were collected at admission.Routine blood tests were done,and then VEGF levels in plasma were measured by enzyme linked immunosorbent assay (ELISA).Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores were recorded every day for 7 days.Patients' prognosis was assessed during the following 28 days.The patients were divided into 28-day survival group and non-survival group.Comparison between two groups was done by single factor analysis.Spearman rank correlation was used to analyze the correlation between VEGF levels and PLT.Mutivariate logistic regression analysis was performed to identify the independent risk factor for 28-day prognosis.Receiver operating characteristic curve (ROC curve) was plotted,and the effect of related indexes on predicting 28-day survival was evaluated by area under ROC curve (AUC).Results There were no significant differences in VEGF (ng/L:471.73 ± 198.34 vs.383.49 ± 266.54,t=-1.918,P=0.057),PLT (× 109/L:220.40±127.60 vs.246.42± 100.72,t=1.275,P=0.204),leucocyte counts (× 109/L:12.48 ±4.62 vs.13.70 ±5.97,t=1.063,P=0.292),mean arterial pressure [mmHg (1 mmHg=0.133 kPa):86.50 ± 12.04 vs.91.03 t 13.10,t=1.557,P=0.123] and blood lactic acid (mmol/L:1.79 ± 1.30 vs.1.50 ± 0.60,t=-1.768,P=0.079) at admission between the non-survival group (n=42) and survival group (n=93).VEGF/PLT (2.59 ± 1.44 vs.1.73 ± 1.13,t=-3.756,P=0.000) as well as APACHE Ⅱ scores (15.50 ± 4.50 vs.13.28 ± 4.61,t =-2.022,P=0.045) of the non-survival group were significantly higher than those of survival group,and oxygenation index (PaO2/FiO2) of the non-survival group was significantly lower than that of survival group (kPa:32.38 ± 11.12 vs.37.04 ± 10.97,t=2.278,P=0.024).Correlation analysis showed that the concentration of VEGF was positively correlated with PLT (r=0.271,P=0.001).It was shown by multivariate logistic regression analysis that only VEGF/PLT was the independent risk factor in predicting 28-day prognosis in patients with sepsis [odds ratio (OR) was 1.591,95％ confidence interval (95％CI) 1.164-2.175,P=0.004].AUC of VEGF/PLT was 0.704 ± 0.047 (P=0.000,95％CI:0.611-0.797) for predicting 28-day survival.The optimal cut-off point was 1.32,and the sensitivity and specificity were 81.0％ and 48.4％,respectively.Conclusion VEGF/PLT can be used as one of the indicators to predict 28-day survival in patients with sepsis.

BACKGROUND:Bone marrow mesenchymal stem cels (BMSCs) have the ability of multi-directional differentiation. Polygonatum sibiricum polysaccharide can promote osteogenetic differentiation of mouse BMSCs by activating Wnt/β-catenin signaling pathway, which is expected to become a new drug for the treatment of osteoporosis.
OBJECTIVE:To investigate the effects of Polygonatum sibiricum polysaccharide on Wnt/β-catenin signaling pathway in the osteogenic differentiation of mouse BMSCs.
METHODS:The mouse BMSCs were cultured and induced in osteoblast medium containing final concentrations (5, 10, 25, 50mg/L) of Polygonatum sibiricum polysaccharide. The mouse BMSCs treated without Polygonatum sibiricum polysaccharide was set as the negative control group. The morphological changes of cels were observed under an inverted microscope. Alkaline phosphatase (ALP) activity assay was performed by PNPP method. The mineralization nodules were observed and stained with alizarin red S and the number and area fraction were recorded under an inverted microscope. The mRNA expressions of osteogenesis-related genes ALP, Runx2, and osteocalcin were evaluated by quantitative real-time PCR (qRT-PCR). qRT-PCR and western blot were used to determine the expression level of β-catenin. The downstream β-catenin/TCF transcriptional activity was evaluated with the Dual-Luciferase Reporter Assay System.
RESULTS AND CONCLUSION: Compared with the control group, polygonatum sibiricum polysaccharide significantly enhanced the alkaline phosphatase activity, the mineralization ability of cels, and the mRNA expression of ALP, Runx2 and osteocalcin in the differentiated BMSCs in a dose dependent manner (P <0.05). After induction, the mRNA expression of β-catenin was the highest on the 3rd day. Polygonatum sibiricum polysaccharide significantly increased the expression of β-catenin (P < 0.05) in the process of promoting the differentiation of BMSCs into osteoblasts, and also promoted the high-level expression of luciferase reporter gene (TOPFlash) which contains wild type TCF binding sites (P < 0.05). These results demonstrate that Polygonatum sibiricum polysaccharide can promote the osteoblast differentiation of mouse BMSCs by activating the Wnt/β-catenin signaling pathway.

Objective To establish a oligochip method for detection of HBV Lamivudine drug resistance and evaluate the clinical role of the chip system.Methods 388 HBV DNA positive sera from patients receiving lamivudine treatment and 559 chronic hepatitis B patients not receiving lamivudine treatment,and 359 sera from HBV DNA negative controls were assayed for HBV mutations utilizing oligonucleotide microarray.Meanwhile,these results were contrasted with Quantitative PCR and DNA sequencing method.Results The results of clinical evaluation shows that for the codons 528,552 and 555,the agreements between the microarray and sequencing data are 96.6%,98.5% and 100%,respectively.In the 559 samples,which were detected positive for HBV DNA by quantitative PCR,all but three weak positive samples were positive by the microarray,demonstrating an agreement of 99.7%.All the 359 HBsAg negative samples were shown to be negative for HBV DNA by the microarray method.Conclusions The HBV-Lamivudine oligochip is eligible to detecting wild type HBV and HBV lamivudine-mutants in patient's sera.It has great potential application of administration for lamivudine treatment in HBV patients.

Objective To investigate the characteristic changes in cerebral infarction and brain edema. Method A total of 122 Healthy adult male Spraque-Dawley rats were randomly (random number) divided into three groups: normal group ( n = 12), sham operated group (n=12) and cerebral ischemia group ( n = 98). Cerebral infarction and brain edema were induced by a permanent occlusion of right middle cerebral artery (POM-CA) with ligature. According to the duration of POMCA, the rats of cerebral ischemia group were further divided into seven sub-groups, 2 h, 4 h, 6 h, 12 h, 18 h, 24 h and 30 hours. The hemispheric ratio was detected by staining with 2% 2,3,5-triphenyltetrazolium chloride solution, and brain water content was assayed by dry/wet ratio 2 h, 4 h, 6 h, 12 h, 18 h, 24 h and hours after POMCA. Results There was a focal cerebral infarction in the rats of cerebral ischemia group 4 hours after POMCA. There was no significant difference in hemispheric ratio between 4 hours and 6 hours after POMCA by One-way ANOVA (P = 0.091). Compared with 6 h sub-group, the hemispheric ratio increased significantly in 12 h, 18 h, 24 h and 30 h sub-groups (P < 0.01), and the peak was in the 24 h sub-group. The brain water content began to increase 4 hours after POMCA and aggravated 6 hours later, and reached the peak 24 hours after POMCA. The brain water content of the non-ischemic hemisphere increased 18 h,24 h and 30 hours after POMCA. Furthermore, there was a significant correlation between the hemispheric ratio and brain water content ( r = 0.834, P < 0.01). Conclusions The critical point of cerebral infarction and brain edema aggravated is 6 hours after POMCA. Both brain edema and cerebral infarction reach the most serious degree 24 hours after POMCA. It is an important experimental evidence for evaluating the milieu conducive to the pathogenesis, and choosing the suitable time window for the treatment of cerebral infarction and brain edema.