Objective To determine the changes of caspase 3 and 8 genes and protein expressions during indomethacin (IND) induced gastric mucosal cell apoptosis in vivo . Methods Healthy male SD rats were treated intragastrically with four different doses of IND. The rats were killed 3 hours after IND administration and the TUNEL technique was applied to detect mucosal cell apoptosis. The change of caspase 3 mRNA expression was detected by in situ hybridization and RT PCR techniques and the changes of caspase 3, 8 protein expression were monitored immunohistochemically. Results In the rats of control group, TUNEL assay revealed that only a few apoptotic cells. Oral administration of IND resulted in the appearance of massive TUNEL positive cells. Computer aided image analysis showed that the mean pixels in 30~120mg/kg groups were 6 3 , 8 0 , 12 6 and 17 1 fold that in control group ( P

Objective To determine the role of endogenous NO in gastric mucosal tolerant cytoprotection under stress and its possible mechanism. Methods SD rats were exposed to WRS repeatedly during which L NAME, a non selective NOs inhibitor, and L Arg, a substrate for NO synthesis, were administered to inhibit or promote the synthesis of NO, GMBF was measured using LDF 3 flowmeter, NO levels in gastric mucosa were tested by Griess reaction and gastric mucosal lesions were evaluated by ulcer index (UI). Results Gastric tolerant cytoprotection was accompanied by increased GMBF and NO levels in gastric mucosa. Inhibition of endogenous NO synthesis by L NAME worsened mucosal lesions induced by WRS. After repeated WRS, adaptive increase of GMBF was abolished and NO content in gastric mucosa significantly reduced. In contrast, enhancement of endogenous NO synthesis by L Arg attenuated mucosal erosions caused by WRS. GMBF and NO content in mucosa increased. After 4th WRS, mucosal lesions could be negligible. Conclusion By regulating GMBF, endogenous NO might play an important role in the gastric mucosal tolerant cytoprotection under stress. Inhibition of NO synthesis delayed the induction of tolerant cytoprotection, while increase NO synthesis will ptomote the induction of tolerant cytoprotection.

SD rats were exposed to single or repeated water immersion restraint stress (WRS) for 4 h every other day for up to 6 days, during which the extent of gastric mucosal lesions was evaluated grossly and histologically. The expressions of EGF and EGFR in gastric mucosa were assayed by immunohistochemistry. The results showed that single exposure to WRS led to haemorrhagic lesions, but tolerance developed following repeated exposures to WRS. Histological evaluations showed that single exposure to stress conditions led to crater like haemorrhagic necrosis almost extending to the muscularis mucosae. The submucosa was apparently congestive and edematous. Following repeated WRS, however, mucosal haemorrhagic necrosis was significantly attenuated and submucosal edema and congestion apparently lessened. Active regeneration and proliferation of mucosal cells were observed at the same time. The expressions of EGF and EGFR could be detected in normal gastric mucosa. EGF was expressed in cytoplasma mainly in the regenerative zone with weaker expression at the basal portions of the gastric glands. EGFR was mainly distributed on the membrane of cells in the regenerative zone. Single WRS led to significant decreased in the expressions of EGF and EGFR. their expressions were absent in the necrotic region. After repeated WRS, enhanced expression of EGF and its receptors could be observed not only in mucosal cells of regenterative zone but also in other areas including the lumen of gastric glands. The results suggested that single WRS could cause severe gastric mucosal lesions, but gastric mucosa became tolerant to repeated WRS by way of toleration cytoprotection. Gastric adaptation was accompanied by active regeneration of mucosal cells mediated by EGF binding with its receptor, EGFR.

To determine the role of GMBF in gastric mucosal tolerant cytoprotection under stress and its possible regulator, SD rats were exposed to repeated WRS, during which L NAME , a non selective NOS inhibitor, or L Arg, a substrate for NO synthesis, was administered to inhibit or promote the synthesis of NO, GMBF was measured using LDF 3 Flowmeter, NO level in gastric mucosa was monitored by Griess reaction, and gastric mucosal lesions were evaluated by UI. The results showed that gastric tolerant cytoprotection was accompanied by increased GMBF and NO level in gastric mucosa. Inhibition of endogenous NO synthesis by L NAME worsened mucosal lesions induced by WRS. After repeated WRS, adaptive increase of GMBF was abolished and NO content in gastric mucosa significantly reduced. In contrast, enhancement of endogenous NO synthesis by L Arg attenuated mucosal erosions produced by WRS and GMBF, NO content in mucosa increased. Good relationships between the changes in GMBF, UI, NO content in mucosa were found. It suggested that GMBF might play an important role in gastric mucosal tolerant cytoprotection. Endogenous NO might be one of its regulators. Inhibition of its synthesis delayed the induction of tolerant cytoprotection, while enhancementpromoted it.

[ ABSTRACT] AIM:To observe the change of skin histology in diabetic rats and to investigate the possible me-chanism of c-Jun N-terminal kinase (JNK) protein in the dorsal root ganglion (DRG) during the process.METHODS:Diabetic animal model was established in the male SD rats by intraperitoneal injection of streptozotocin.Plantar skin speci-mens of the rats were collected from control group, DM 2-week group (DM2), DM 4-week group (DM4), and DM 8-week group ( DM8) .Immunohistochemical staining and HE staining were used to observe the change of PGP 9.5 immunoreactive nerve terminals and the structures of the skin tissues.The protein expression of PGP 9.5 in the plantar skin tissues, and JNK and p-JNK protein in the DRG within lumbar 5, 6 (L5, 6), and sacral 1 (S1) spinal cord segments were detected by Western blotting.RESULTS:PGP 9.5 immunoreactive nerve terminals of the plantar skin of the rats mainly distributed in the basal layer of the epidermis and papillary dermis.Compared with control group, PGP 9.5 positive nerve terminals in DM4 group showed reduced density and sparse distribution.PGP 9.5 positive nerve terminals in DM8 group showed signifi-cantly reduced distribution, thinner nerve diameter, shorter length and distorted shape.Histological changes of the thinner epidermal tissue, reduced epidermal cell layers, uneven cell distribution and arrangement in DM4 group, and significantly reduced epidermal cell layers, swollen and blurred cells, increasing cell gap, lack of stratified epidermis arrangement for part of epidermis, atropal and degenerated dermal collagen fiber, significantly decreased subcutaneous fat in DM8 group were observed.The results of Western blotting showed that the protein expression of PGP 9.5 in the plantar skin tissue of DM rats was progressively decreased along with the disease, while the protein level of p-JNK in L5, 6-DRG or S1-DRG showed a gradual increasing trend.PGP 9.5 immunoreactive positive nerve terminal density of plantar skin in DM rats had a negative correlation with the protein level of p-JNK in L5, 6-DRG and S1-DRG (P<0.01), but showed a significant positive correlation with the plantar skin thickness (P<0.01).CONCLUSION:The protein level of p-JNK within L5, 6-DRG or S1-DRG in DM rats shows a progressive enhancement.At the same time, there is a significant change in the skin tissue density and structure.The changes of skin tissue and nerve morphology in DM rat may be related to the activation of JNK/SAPK pathway in L5, 6-DRG or S1-DRG cells.Blocking or inhibiting JNK/SAPK pathway may delay the diabetic pe-ripheral neuropathy and reduce the risk of skin lesions.

AIM:To investigate the effects of ginsenoside Rg1 on the behavioral changes and the autophagy of hippocampal neurons of the rats with post-traumatic stress disorder (PTSD).METHODS:The Sprague-Dawley rats were randomly divided into 5 groups:control group, model group, fluoxetine group, low-dose ginsenoside Rg1 group and high-dose of ginsenoside Rg1 group.The combination of single prolonged stress and foot stock was performed to induce PTSD-like animal model.The rats in fluoxetine group was administered with fluoxetine by gavage at dose of 10 mg/kg for 21 d, while the rats in low and high doses of ginsenoside Rg1 groups were administered with ginsenoside Rg1 by gavage at doses of 20 mg/kg and 40 mg/kg for 21 d, respectively.The rats in control group and model group were both given saline by gavage for 21 d.The open-field test and stiff behavior test were used to examine the behavioral changes of the rats.The morphological structure and numerical changes of the hippocampal neurons were observed by Nissl-staining method.We adopted immunofluorescence labeling to observe the beclin 1 and LC3 positive hippocampal neurons and the levels of beclin 1 and LC3-Ⅱ/LC3-Ⅰratio in rat hippocampus.RESULTS:Compared with control group, decreased vertical movement time and horizontal movement time in open-field test and increased rate of stiff behavior in the stiff behavior test were observed in model group.Hippocampal neurons in model group were loosely arranged with vacuole-like structures and different degrees of cell shrinkage in contrast with control group.More beclin 1 and LC3 positive cells were identified, and higher protein levels of beclin 1 and ratio of LC3-Ⅱ/LC3-Ⅰ in model group were found as compared with control group.However, increase in movement in open-field test and decrease in stiff behavior were detected in the rats treated with low-and high-dose ginsenoside Rg1 as compared with the model rats.Meanwhile, vacuole structures, the numbers of beclin 1 and LC3 positive neurons, the protein expression of beclin 1 and LC3, and the total cell numbers were increased.Higher dose of ginsenoside Rg1 had more profound effects on these observed results.CONCLUSION:Ginsenoside Rg1 alleviates the abnormal behaviors in the PTSD rats, which might be related to the inhibition of abnormal autophagy of hippocampal neurons.

OBJECTIVE:To establish a method for the determination of sodium valproiate in human serum and to study the bioequivalence of steady state concentration C ssm in of the domestic and imported sodium valproiate sustained-release compound tablets.METHODS:Two periods of multi-oral administration of domestic and imported sodium valproiate sustained-release compound tablets were conducted alternately at random on20healthy male volunteers;the trough concentration of sodium valproate in human serum was determined by HPLC-fluorescence detection and the data were analyzed by3p97pro?gram.RESULTS:The blood concentration was steady after3d oral administration of both the domestic and imported sodium valproate sustained-release compound tablets.The C ssm in of domestic and imported products were(38.17?9.36)、(35.48?9.44)mg/L respectively.C ss min of domestic and imported sodium valproate sustained-release compound tablets were of bioe?quivalence either single or multi-oral administration.CONCLUSION:This HPLC-fluorescence method is quick,sensitive and economical,which can be used to monitor the concentration of sodium valproate in human serum.

Objective To analyze the effect of autologous bone marrow stem cells transplantation in treatment of patients with decompensated cirrhosis. Methodls Seventy-eight patients (aged from 26 to 67) with decompensated cirrhosis, including 56 with hepatitis B, 21 with alcoholic cirrhosis and 1 with schistosomial cirrhosis, were included. Bone marrow was aspirated from poster superior spine. After isolation and purification, the stem cells were transplanted into liver via hepatic artery. The liver function, laboratory parameters and Child-Tureotte-Pugh scores were evaluated in 2,4 and 8 weeks after transplantation. Results At the 4th week after transplantation, the level of albumin was increased obviously from (32.9±5.58) g/L to (38.32±6.45) g/L,whereas the alanine aminotransferase was decreased from (96.92±83.91) U/L to (73.48±18.46)U/L. It was revealed that the prothrombin time was decreased from (16.66±3.91) s to (15.52±3.35) s and fibronegen increased from (2. 22 ± 0. 88) g/L to (2. 58±0. 88) g/L. After transplantation, appetite was improved in 72 cases (92.3%), ascites was decreased in 70 cases (89.7%) and abdomen distention was ameliorated in 68 cases (87.2%). There was no complications related to the transplantation. Conclusion Transplantation of autologous bone marrow stem cells is a safe and effective method in treatment of patients with decompensated cirrhosis.

Objective In recent years , multivariate pattern analysis ( MVPA) method was proposed and considered to be a promising tool for automated identification of various neuropsychiatric populations .Support vector machine ( SVM) is one of the most widely used methods of MVPA .Using SVM classifier for MVPA of amnestic mild cognitive impairment (aMCI) and normal control (NC) group, the present study aims to build an individual diagnostic model with significant discriminative power and investigate the gray matter abnor-malities of aMCI patients . Methods Fifty-one aMCI patients and 68 normal controls were scanned on the 3-Tesla magnetic resonance imaging (MRI) for high-resolution T1-weighted images.Gray matter volume map was calculated for each subject and used as features for subsequent discriminative analysis .We first applied feature selection to remove redundant information and reduce feature dimension , and then trained an SVM classifier . Leave-one-out cross validation ( LOOCV) was used to estimate the performance of the classifier , and finally the most discriminative features were identified . Results The proposed classifier achieved a classification accuracy of 83.19%with a sensitivity of 76.47%and a specificity of 88.24%.In ad-dition, the area under the receiver operating characteristic (ROC) curve was 0.8368.Further analysis revealed that the most discrimi-native features for classification included bilateral parahippocampal gyri , bilateral hippocampi , bilateral amygdala , bilateral thalamus , right cingulate , right precuneus , left caudate , left superior temporal gyrus , left middle temporal gyrus , left insula and left orbitofrontal cortex. Conclusion The proposed classification model has achieved significant accuracy for aMCI prediction , and it also displayed the whole brain gray matter atrophy pattern in aMCI patients .It suggests that the proposed method may have important implications for early clinical diagnosis of aMCI patients .

The simulator testing in vitro and computational simulation of the artificial knee joint wear are important methods to evaluate the wear performance of the prosthesis in vitro and to predict the clinical performance of knee joint products. Based on the method of literature search, this paper compares the mechanical and kinematic loading input curves carried out by Chinese scholars in recent years, standard curves, and Chinese measurement curves of two typical movements of gait. Data of vitro simulator test and computational simulation model are compared, summarized, and analyzed. The results show that the measured data of motion and load cannot be directly used as the loading conditions for the simulator wear test and computational simulation. The mechanics and kinematics data of Chinese people are different from the international standards. The domestic artificial knee joint in vitro simulator wear test methods are similar but the results of different test institutions are somewhat different. The computation wear prediction research is basically synchronized with foreign countries, but the problem that the calculated wear results are lower than that in vitro test is still unsolved. The artificial knee joint wear performance evaluation system based on Chinese knee joint mechanics and kinematics data is the forward direction of the research.