The human brain-derived neurotrophic factor (hBDNF) gene was cloned by polymerase chain reaction and the recombinant adeno-associated viral vector inserted with hBDNF gene (AAV-hBDNF) was constructed. Cultured rat hippocampal neurons were treated with Abeta(25-35) and serued as the experimental Abeta-induced neuronal damage model (AD model), and the AD model was infected with AAV-hBDNF to explore neuroprotective effects of expression of BDNF. Cell viability was assayed by MTT. The expression of bcl-2 anti-apoptosis protein was detected by immunocytochemical staining. The change of intracellular free Ca ion ([Ca2+]i) was measured by laser scanning confocal microscopy. The results showed that BDNF had protective effects against A-induced neuronal damage. The expression of the bcl-2 anti-apoptosis protein was raised significantly and the balance of [Ca2+]i was maintained in the AAv-hBDNF treatment group as compared with AD model group. These data suggested that recombinant AAV mediated a stable expression of hBDNF in cultured hippocampal neurons and resulted in significant neuron protective effects in AD model. The BDNF may reduce neuron apoptosis through increasing the expression of the bcl-2 anti-apoptosis protein and inhibiting intracellular calcium overload. The viral vector-mediated gene expression of BDNF may pave the way of a novel therapeutic strategy for the treatment of neurodegenerative diseases such as Alzheimer's disease.

ObjectiveTo investigate the clinical value of serum complement C3 and C4 levels for predicting the severity of hepatic fibrosis in patients with chronic hepatitis B.MethodsHistopathological diagnosis was confirmed in 442 patients with chronic hepatitis B.Serum complement C3 and C4 levels were determined by Beckman-Coulter Immage 800 immunochemistry system.ROC curve was used to analyze the value of serum complement C3 and C4 levels in predicting the severity of hepatic fibrosis.ResultsThe areas under ROC curve of complement C3 and C4 for predicting significant fibrosis ( ≥ S2),severe fibrosis ( ≥ S3) and cirrhosis (S4) were all significantly larger than the area under diagonal reference line ( P =0.009,0.000,0.000 and P =0.005,0.000,0.000,respectively).According to ROC curves,the optimal cut-offs of serum complement G3 for predicting severe fibrosis and cirrhosis were ≤0.74 g/L and ≤0.64 g/L,and the corresponding sensitivity,specificity,positive predictive value,negative predictive value,accuracy were 0.585,0.681,0.617,0.650,0.636 and 0.509,0.775,0.423,0.830,0.710,respectively.The optimal cut-offs of serum complement C4 for predicting severe fibrosis and cirrhosis were ≤0.14 g/L and ≤0.12 g/L,and the corresponding sensitivity,specificity,positive predictive value,negative predictive value,accuracy were 0.565,0.634,0.576,0.623,0.602 and 0.463,0.781,0.407,0.818,0.704,respectively.ConclusionSerum complement C3 and C4 may be used for predicting severe fibrosis and cirrhosis in patients with chronic hepatitis B,but its stability and reliability need to be improved.

Objective To study the effects of rhubarb on the mitogen-activated protein kinase (MAPK) sig-naling transducfon pathway in rats with severe acute pancreatitis (SAP),and to investigate the treatment mecha-nism of rhubarb on SAP. Method One hundred SD rats were provided by from the Animal Center of Nanjing Uni-versity. All animals were randomly divided into sham operation (n=33), SAP (n=33) and rhubarb groups (n=34). SAP model was induced by retrograde injection of 5% sodittm taurocholate. Rhubarb was given with 10% rhubarb decoction (2 mi/100 g) at the time of pancreafitis induction in the rhubarb groups. At 1, 3, 6, and 12 h after the models were established,animals were killed. MAPK activity in pancreatic tissue was examined by West-em blotting and the mRNA levels of TNF-α and IL-6 in pancreatic tissues were detected by RT-PCR. All data were analyzed by SPSS statistical software and statistical differences between values from two sroups were determined by the Student's t -test. Results MAPK activity, TNF-α and IL-6 mRNA levels in pancreatic tissues were signifi-cantly enhanced in the SAP group compared with the sham operation group (all P<0.01). Rhubarb treatment markedlyinhibited MAPK activation,TNF-α,IL 6 mRNA (all p<0.01). Conclusions Rhubarb can alleviate the inflammatory response of SAP by down-regulating MAPK activity.

Objective To investigate the effect of propentofylline on nerve growth factor (NGF) and IL-1βrelease from rat cerebral cortical astrocytes. Methods Primary cultured rat astrocytes from SD rats (1-3 d,weighing 6-8 g) after 4 passages were randomly divided into 8 groups ( n = 6 wells each): group Ⅰ control (group C); group Ⅱ , Ⅲ, Ⅳ the astrocytes were exposed to propentofylline 10, 100 and 1000 μmol/L respectively (group P1, P2, P3 ); group Ⅴ the astrocytes were exposed to LPS 1 μg/ml and group Ⅵ, Ⅶ, Ⅷ the astrocytes were exposed to propentofylline 10, 100 and 1000 μmol/L in addition to LPS 1 μg/ml (group P1 + LPS, P2 + LPS,P3 + LPS). The astrocytes were then incubated for 3 days in all 8 groups. The concentrations of IL-1β and NGF in the supernatant were detected at 1 and 3 days of incubation using ELISA. Results LPS activated astrocytes resulting in decrease in NGF release and increase in IL-1β release. Propentofylline significantly increased NGF release and decreased IL-1β release from astrocytes incubated alone or with LPS by suppressing activation of astrocytes. Conclusion Propentofylline can enhance NGF release and inhibit IL-1β release from rat cerebral cortical astrocytes.

Aim To investigate the effect of intrathecal injection of fluorocitrate(Fc)on mechanical and thermal hyperalgesia induced by complete Freund's adjuvant(CFA)injection in rats.Methods The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured before and after CFA or Fc treatment.The changes of glial fibrillary acidic protein(GFAP)and OX-42(a microglial marker)expression in the spinal cord dorsal horn were evaluated by immunohistochemistry analysis.Results Rats with CFA-induced arthritis showed mechanical allodynia and thermal hyperalgesia,which was correlated with the increased GFAP and OX-42 expression in the spinal cord dorsal horn.Intrathecal injection of Fc markedly suppressed CFA-induced thermal hyperalgesia and mechanical allodynia.Fc significantly attenuated the activation of GFAP and OX-42 in the spinal cord dorsal horn.Conclusions The glia activation in spinal cord is closely related to the progress of CFA-induced peripheral hyperalgesia.Fc may exert antihyperalgesic effect by inhibiting the activation of astrocyte and microglia.

Objective To investigate the current status of type 2 diabetic patients who failed to achieve the glycemic control target, and provide theoretic evidences for making corresponding strategies. Methods The 2 diabetic patients who failed to reach the glycemic target were recruited from 181 hospitals in 26 cities and received a standard questionnaire, the conditions of their blood glucose level, lifestyle intervention, blood sugar monitoring, and drug therapy were recorded. Totally 3 861 questionnaires with complete information were collected. And the causes which account for glycemic control status were analyzed. Results Among these patients, the mean HbA1c was 7.9%, the mean fasting plasma glucose was 8.2 mmol/L, and the mean postprandial plasma glucose was 11.5 mmol/L. Only 25.6% of patients take their diet control strictly as prescribed and 44. 5% of patients have little exercise. 35. 8% and 47.8% of patients did not monitor their fasting and postprandial plasma glucose,respectively. Glycemic control in the patients aged ＞ 60 years was similar to the younger patients, but the hypoglycemia incidence in the elder group reached 35.5%, which was higher than those in the other 2 groups (20.8% and 21.4%, both P＜0. 05 ). The proportion of patients with mono-therapy and combination therapy was 46. 1% and 51.7%, while the proportion with combination therapy rose in the patients aged ＞60 years (58.7%;Compared with the other age-groups, all P＜0.05 ). 75 % of patients have adjusted their drug administration regimen since initial treatment. Conclusions Inadequate or inappropriate drug therapy regimen is a major cause responsible for this poor glycemic control status. In addition, the unhealthy life styles, insufficient blood sugar monitoring, and poor compliance were also important causes. Thus, for these patients, it is necessary to further enhance patients' education, to improve life style intervention, as well as to select more effective, safer, and compliant drug therapy regimens. Finally, the glycemic control target for the elder patients should be more flexible.

The human brain-derived neurotrophic factor (hBDNF) gene was cloned by polymerase chain reaction and the recombinant adeno-associated viral vector inserted with hBDNF gene (AAV-hBDNF) was constructed. Cultured rat hippocampal neurons were treated with Aβ25-35 and serued as the experimental Aβ-induced neuronal damage model (AD model), and the AD model was infected with AAV-hBDNF to explore neuroprotective effects of expression of BDNF. Cell viability was assayed by MTT. The expression of bcl-2 anti-apoptosis protein was detected by immunocytochemical staining. The change of intracellular free Ca ion ([Ca2+]i) was measured by laser scanning confocal microscopy. The results showed that BDNF had protective effects against Aβ-induced neuronal damage. The expression of the bcl-2 anti-apoptosis protein was raised significantly and the balance of [Ca2+]i was maintained in the AAV-hBDNF treatment group as compared with AD model group. These data suggested that recombinant AAV mediated a stable expression of hBDNF in cultured hippocampal neurons and resulted in significant neuron protective effects in AD model. The BDNF may reduce neuron apoptosis through increasing the expression of the bcl-2 anti-apoptosis protein and inhibiting intracellular calcium overload. The viral vector-mediated gene expression of BDNF may pave the way of a novel therapeutic strategy for the treatment of neurodegenerative diseases such as Alzheimer's disease.

Objective To evaluate the role of gliocytes in the spinal cord in the development of inflammatory pain (IP) in rats. Methods Adult male SD rats weighing 180-220 g were used in this experiment. A catheter was implanted in the subarachnoid space according to the method described by Yang. Animals with abnormal motor function of the hindlimb after intrathecal (IT) catheter implantation were excluded. IP was induced by subcutaneous (sc) injection of complete Freund's adjuvant (CFA) 50 μl at the lateral side of the ankle joint of the right hindpaw. Sixty-five rats were randomly divided into 5 groups ( n = 13 each): group I IP control normal saline (NS) 50μl was injected sc instead of CFA; group II IP; group IE PC (IT) + IP control fluorinated citric acid (FC, a gliocyte metabolism inhibitor) 1 nmol/10μl was injected IT at 15 min before NS 50 μl sc injection; group IV NS (IT) + IP and group V FC (IT) + IP. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured 2 d before induction of IP (T_0, baseline) .before and at 2, 4, 6, 8, 10, 12, 24 and 26 h (T_(1-9)) after sc NS or CFA injection. Five enimals in each group were killed at T_5 (8 h after sc NS/CFA injection) and the lumbar segment (L_(4,5)) was removed for determination of glial fibrillary acidic protein ( CFAP) and OX-42 expression by immuno-histochemistry. Results In group Ⅱ and Ⅳ sc CFA significantly decreased MWT and TWL. Mechanical and thermal hyperalgegia induced by sc CFA was significantly suppressed by intrathecal FC in group V . IP significantly increased GFAP and OX-42 expression in the spinal cord. Intrathecal FC significantly attenuated IP-induced up-regulation of GFAP and OX-42 expression in the spinal cord. Conclusion The activation of gliocytes in the spinal cord is involved in the development of CFA-induced hyperalgesia in rats.

Retinal vein occlusion (RVO) is characterized by obstruction of retinal vein blood flow, distended flexion, retinal hemorrhage, edema, and neovascularization, and its pathogenesis is not completely clear. Recent studies have found that endothelin (ET)-1, ETA receptor and ETA signaling pathways in the ET system may be involved in the occurrence and development of RVO by stimulating vasoconstriction to increase retinal vein pressure and inducing the expression of pro-inflammatory factors such as TNF-α, IL-6 and IL-1β. In-depth understanding of the correlation between the ET system and the occurrence and development of RVO can provide new ideas for further research on the pathogenesis of RVO.

OBJECTIVETo analyze the efficacy and safety of laparoscopic adjustable gastric placation (LAGBP), a new procedure for surgical treatment of obesity.

METHODSClinical and 1-year follow-up data of 10 patients who underwent LAGBP in our department between September and November 2011 were analyzed retrospectively.

RESULTSThe mean operative time was (93.0±13.4) min, while the mean intraoperative blood loss was (15.5±4.7) ml. The mean excessive body weight loss rate(%EWL) at 3, 6, 9 and 12 months after the operation was 25.1%, 40.6%, 45.3% and 50.8% respectively. There were no severe post operative complications.

CONCLUSIONSLAGBP is associated with high safety and good short-term efficacy.