Objective To investigate the mechanism of preconditioned neuroprotection of cerebellar fastigial nucleus electrical stimulation(FNS) by means of observation of the effect of FNS on the expressions of PKC? and PKC? in the somatosensery cortex and basal ganglia in rats. Methods FNS models in rats were established. Brain tissue containing the somatosensery cortex and basal ganglia was removed at 0 h and 1, 3, 7 and 10 d respectively after FNS and 20 ?m coronal sections were obtained with a sliding cryo microtome. Immunocytochemical analysis of PKC? and PKC? in 2 sections from each rat was performed and the average optical density of immunostaining was quantified by computer assisted image analysis system. Sham stimulation rats and cerebral dentate nucleus (DN) stimulation rats were used as the control. Results At 0 h after FNS, no significant changes of the expressions of PKC? and PKC? were found in the contralateral somatosensery cortex and basal ganglia. The expressions of PKC? and PKC? increased significantly at 1 d and decreased at 3 d but were still higher than those in sham stimulation group at 7 d and decreased to the basal level of the control at 10 d. However, the expressions of PKC? and PKC? in the ipsilateral somatosensery cortex and basal ganglia were also elevated at 1 d after FNS but the increase was significantly lower than that in the contralateral ones and reached the basal level at 3 d. There was no change of PKC? and PKC? in sham stimulation group at 1 d after DN stimulation. Conclusion The increased expressions of PKC? and PKC? in the somatosensery cortex and basal ganglia induced by FNS may participate in the preconditioned neuroprotection of FNS.

Objective To investigate the change of plasma cholinesterases (CHEs) in the people with diabetes or fatty liver or overweight , and explore the role of CHE in these diseases .Methods The plasma CHEs in 2834 subjects were detected , and these subjects were divided into five groups , including diabetes , fatty liver , overweight , diabetes with fatty liver , and the normal groups . Results The plasma CHE activities in diabetes group , fatty liver group , overweight group , and diabetes with fatty liver group were all higher than the normal group [(8943 ±1896)U/L, (9716 ±1673)U/L, (8798 ±1710)U/L, (9385 ±1687)U/L vs (8028 ±1621) U/L], and the CHE level in the fatty liver group was highest among five groups .However, the CHE level in diabetes group or fatty liv-er group was not significantly different from that in the diabetes with fatty liver group .The CHE level of the people with components of metabolic syndrome (MS) was significantly higher than that without MS component [(8786 ±1514)U/L, (9141 ±1771)U/L, (9705 ±1628)U/L, (9138 ±1768)U/L, (9530 ±1607)U/L vs (7821 ±1324)U/L]),but the CHE level was not increased gradually with the increased MS component.The plasma CHE had a negative correlation with age ( P =0.00),but it had a positive correlation with triglyceride (TG), total cholesterol (TC), and body mass index (BMI)( P =0.00).Conclusions The plasma CHE activity was el-evated in diabetes group , fatty liver group , and overweight group , which might be a risk factor in these diseases .Controlling the plas-ma CHE might help to treat the metabolism diseases .

Objective To investigate the effects of cyclooxygenase-2 (COX-2) selective inhibitor combined with 5-lipoxygenase (5-LOX) inhibitor docebenone (AA861) on cell proliferation and migration of human colon cancer cell line HT-29. Methods Cultured in vitro of HT-29 cells in high metastatic colon cancer, A A861 and celecoxib on colon cancer cell drugs for 48 h, the cell proliferation was assayed by MTT; the migration of cell was detected by Transwell; immunofluorescence staining of intracellular changes in ICAM-1 protein, RT-PCR detect the gene expression of ICAM-1 and VEGF. Results MTT and Transwell experiments showed that the inhibition on celecoxib and AA861 on colon cancer was dose-dependent and time-dependent. And the inhibition of AA861 and celecoxib 100 μmol/L alone on colon cancer cells proliferation were 43.2 % and 42.8%, and when the two drugs 100 μmol/L combined on colon cancer cells the inhibition was 53.8%, and the difference between alone group and combined group was statistical (P ＜0.001). The inhibition of AA861 and celecoxib 100 μmol/L alone on colon cancer cells migration were 32.0 % and 29.3%, and when the two drugs 100 μ,mol/L combined on colon cancer cells the inhibition is 57.8 %, and the difference of between alone group and combined group was statistical (P ＜0.001). Immunefluorescence staining and RT-PCR results suggested that the two drugs can inhibit ICAM-1 and VEGF protein and gene expression, and when the two drugs combined, a stronger inhibition effect appeared than used alone (P＜0.001). Conclusion Low-dose celecoxib and AA861 combined has a synergistic inhibited effect on colon cancer cells invasion and metastasis, and the mechanism relates with the VEGF and ICAM-1 expression.

Objective To study the effect of 5-lipoxygenase(5-LOX) inhibitor nordihyroguaiaretic acid (NDGA) combined the selective cyclooxygenase-2 (COX-2) inhibitor Celecoxib on the apoptosis of human colon carcinoma cell line HT-29. Methods Different concentration of NDGA and Celecoxib combinations were used to process cancer cell, and thiazolyl blue tetrazlium bromide (MTT) and phase contrast microscope and Annexin V/PI fluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to study the proliferation inhibited effect and apoptosis induced effect caused by combination of NDGA combined Celecoxib. Results MTT results showed that the viability of NDGA group, Celecoxib group and the group of NDGA combined Celecoxib (0.432±0.024,0.425±0.013,0.303±0.014 vs 0.693±0.018,t＝18.79,25.75,37.64,P＜0.01) was obviously lower than control group. The group of NDGA combined Celecoxib was significantly lower than NDGA group or Celecoxib group (t＝10.21, 14.14,P＜0.01). Under inverted phase contrast microscope, cell morphology significantly changed, and the group of NDGA combined Celecoxib changed most obviously. Apoptosis was observed by laser scanning confocal microscope (LSM) after NDGA and Celecoxib were used to process the HT-29. RT-PCR showed that up-regulation of Caspase-3 after treatment, and the combination of two drugs increased the most. Conclusions NDGA combined Celecoxib inhibited proliferation and induced apoptosis in human colon carcinoma cell line HT-29, and combined therapy had better effect than that of any drug used separate-ly. The mechanism may be associated with up-regulation of Caspase-3.

Nuclear factor κB (NF-κB) is an important cellular transcription factor. The important role of NF-κB-mediated cell signal transduction pathway in apoptosis is a hot topic at home and abroad. In order to discover new regulators in NF-κB signaling pathway, a high-throughput cell-based screening model based on dual luciferase reporters system was established, a number of genes that can activate NF-κB signal pathway were obtained by screening of 439 novel function genes. Among them, TMEM9B can obviously activate NF-κB signaling pathway. Further experiments showed that TMEM9B activated NF-κB signaling pathway in a dose-dependent pattern. Western blotting and EMSA experiments confirmed that TMEM9B can promote the degradation of IκBα (a cytoplasm inhibitor of NF-κB), and cause NF-κB shift from the cytoplasm to nucleus. At the same time, flow cytometry result demonstrated TMEM9B can induce apoptosis in HEK293T and HeLa cells. In short, a stable and effective screening system for NF-κB has been established, through which TMEM9B was identified to be able to significantly activate NF-κB signal transduction pathway and thus cause cells apoptosis.

Objective To investigate the correlation of the expression of KAI1/CD82 and laminin receptor (LNR) in cholangiocarcinoma, and study its role in the invasion and metastasis of cholangiocarcinoma. Methods The expressions of KAI1/CD82 and LNR in 48 cholangiocarcinoma tissue samples were detected by SP immunohistochemistry, and their relationships with clinicopathological factors were analyzed. Results The positive expression rates of KAI1/CD82 and LNR in cholangiocarcinoma were 31% (15/48) and 54% (26/48), respectively. In highly differentiated cholangiocarcinoma, the positive expression rate of KAI1/CD82 was high (χ2=3.911, P<0.05), while that of the LNR was low (χ2=6.970, P<0.05). The positive expression rate of KAI1/CD82 in cholangiocarcinoma with metastasis was significantly lower than that in cholangiocarcinoma without metastasis (χ2=5.765, P<0.05), while the positive expression rate of LNR in cholangiocarcinoma with metastasis was significantly higher than that in cholangiocarcinoma without metastasis (χ2= 9.952, P<0.05). The expression level of KAI1/CD82 was negatively correlated with that of the LNR ( r = -0.462, P < 0.01 ). Conclusions The up-regulated expression of LNR in cholangiocarcinoma correlates with the decreased expression of KAI1/CD82, and plays an important role in the invasion and metastasis of cholangio-carcinoma.

Objective To observe the genetic predisposition of complement C5 gene polymorphisms in proliferative diabetic retinopathy (PDR) in Chongqing Han population.Methods 400 type 2 diabetes (T2D) patients (case group) and 600 age-and sex-matched healthy controls (control group) were enrolled in this study.There were 8 PDR patients in case group.All the subjects were Han ethnic people.The immune-related representative SNP locus of C5 gene including rs2269067,rs7040033,rs7027797 were screened by linkage disequilibrium analysis.Locus rs1017119 was selected by TagSNP and was around the above three loci.Subjects' peripheral venous blood was collected and DNA was extracted.Genotyping was examined by PCR-restriction fragment length polymorphism method.The level of C5 plasma protein was measured by enzyme-linked immunoabsorbent assay.Results The frequency of GG genotype of rs2269067 was significantly increased in PDR patients in cases group compared with controls (Pc=3.4 × 10-5,OR=1.87,95％CI=1.43-2.44;P=3.1 × 10-6).There was no differences in frequency of G,CC and CG genotype of rs2269067 between two groups (P=1.4 × 10-4,1.000,1.0 × 10-6).There were no differences in frequency of G,CC,CG,GG genotype of rs7040033,rs1017119,and rs7027797 between two groups (P＞0.05).The production of C5 plasma protein was significantly increased in case group as compare with control group (P=0.0004).An increased production of C5 plasma protein was observed in rs2269067 GG genotype cases compared to CG or CC cases (P=0.003,0.001).Conclusion C5 rs2269067 GG genotype may be associated with the PDR of T2D in Chongqing Han population.

Objective To summarize the causes of abdominal pain in the early postoperative period after laparoscopic cholecystectomy(LC), so as to employ preventative measures. Methods The clinical data of 35 patients with early postoperative abdomial pain after LC were analyzed retrospectively and the literature was reviewed . Nine cases underwent reoperation, and conservative treatment was successfully performed in the other 26 cases. Results Traumatic factors were involved in 8 cases, including 6 cases of postoperative biliary fistula , and 2 cases with calculus retained in the trocar hole of the abdominal wall. 27 cases had no traumatic factor and included 2 cases of ascariasis of common bile duct, 5 cases with calculus of common bile duct, 1 case of acute pancreatitis, 17 cases of bile duct dysfunction, 1 case of duodenal ulcer and 1 case of stomach cancer. All were cured. Conclusions The key to decrease complications after LC is a complete examination before LC and pay attention to each aspect of perioperative management.

AIM: To optimize the extraction of Weining-Dispersible Tablet(Radix et Rhizoma notoginseng,Radix et Rhizoma ginseng) with the help of central composite design-response surface methodology. METHODS: The main influencing factors of extraction effect included the quantity of ethanol,the concentration of ethanol and extraction time and the extraction conditions were evaluated by the extract yield and the contents of total notoginseng saponins.The optimal extraction of Weining-Dispersible Tablet was selected by central composite design-response surface methodology. RESULTS: The optimal conditions consisted of extraction two times,2 h each time with ten fold of 70% ethanol. CONCLUSION: It is simple,convenient and high predictive to optimize the extraction craft of Weining-Dispersible Tablet by central composite design-response surface methodology.