Objective To investigate the role of tet methylcytosine dioxygenase 2 (TET2) in the regulation of transforming growth factor-β1 (TGF-β1) expression in human glomerular mesangial cells induced by high glucose.Methods Cultured human glomerular mesangial cells were divided into normal control group (5.5 mmol/L glucose) and high glucose group (30.0 mmol/L glucose) which was cultured for 12 h to 72 h.The gene expression of TET2 in mesangial cells were inhibited by small molecule chemical called SC1,and which were divided into high glucose group (30.0 mmol/L glucose+ DMEM),DMSO group (30.0 mmol/L glucose+0.1％DMSO) and SC1 group (30.0 mmol/L glucose+3 μmol/L SC1).The mRNA and protein expression of TGF-β1,TET1 to 3 and α-smooth muscle actin (α-SMA) was detected by quantitative real-time PCR and Western blotting.Methylation of CpG islands in the regulation region of TGF-β1 was detected by bisulfite sequencing PCR (BSP).The activity of mesangial cell proliferation was assessed by colorimetry of thiazolyl blue (MTT).Results Compared with normal control group,the mRNA and protein expression of TET2 in mesangial cells induced by high glucose was increased significantly in a time-dependent manner (all P ＜ 0.05),but the expression of TET1 and TET3 was not affected.Meanwhile methylation rate of 4 CG sites from 24 h to 72 h were decreased in the first exon of TGF-β1 (P ＜ 0.01),but not in the promoter.Compared with high glucose group,when the expression of TET2 was inhibited by SC1,the methylation rate of TGF-β1 was recovered evidently (P ＜ 0.05),the mRNA and protein expression of TGF-β1 and α-SMA was suppressed,and the proliferation of mesangial cells was decreased (all P ＜ 0.05).Conclusions Demethylation of the CpG island mediated by TET2 may play an important role in the expression of TGF-β1 and mesangial cell phenotype transformation induced by high glucose.

Objective To observe the effects of ketamine on the patients with depression re-ceiving modified electric conulsive therapy (MECT).Methods Sixty patients with depression were randomly divided into ketamine group and propofol group (n =30 each group).Atropine 0.5-1.0 mg, propofol 1.0 mg/kg or ketamine 0.8 mg/kg i.v.were given before MECT,Scoline 0.7-1.0 mg/kg i. v.was given after the eyelash reflex disappeared.Hamilton Depression Rating Scale (HAMD)was completed after the 2 nd ,4 th and 6 th MECT,the time of convulsion,twitch index,energy percentage, respiratory recovery time and adverse reactions were recorded.Results The total score of HAMD was significantly decreased with the increasing times of MECT in both groups,compared with propo-fol group,ketamine group's HAMD total score decreased faster,especially after the 4th MECT,the score decreased significantly in ketamine group (P <0.05).The time of convulsion,twitch index,en-ergy percentage, respiratory recovery time, adverse reactions all had no statistical significance between the two groups.Conclusion Compered with propofol,ketamine,as an anesthetic of MECT, can effectively lower the score of HAMD.

Objective To investigate the effect of dexmedetomidine on excitatory aminoacid (EAA)and inhibition of amino acid(IAA)in cerebro-spinal fluid(CSF)of patients undergoing in-tracranial tumor surgery,and to explore the cerebral protective mechanism of dexmedetomidine in neurosurgery.Methods Sixty patients aged 18-64 years old,ASA Ⅰ or Ⅱ,weighing 50-90 kg un-dergoing elective intracranial tumor surgery were randomly divided into dexmedetomidine group (group D)and control group(group C).Dexmedetomidine 1 μg/kg was infused before anesthesia in-duction for more than 10 minutes and pumped continously with 0.2-0.7 μg·kg-1·h-1 in group D, while in group C midazolam 0.03-0.05 mg/kg was injected followed by intermittent administration of 0.03-0.05 mg/kg.BIS value was maintained between 40-50.MAP and HR was recorded at the time points before induction(T0 ),dura mater incision(T1 ),tumor resection(T2 ),at the end of the surgery (T3 ).And we collected CSF at T0 ,T3 ,6 hours after the surgery(T4 ),12 hours after the surgery (T5 ),24 hours after the surgery(T6 ),then the concentrations of EAA and IAA were determined with high-performance liquid chromatography (HPLC)at T0 ,T3 ,T4 ,T5 and T6 .Results The MAP and HR in group D at T1-T3 were much lower than that in T0 and in group C(P <0.05).Compared with T0 ,the Glu and Asp in CSF significantly increased in group C at T3-T6 and were much higher than those in group D (P < 0.05 ),GABA was significantly decreased and much lower than group D(P <0.05).Compared with T0 ,the Glu and Asp in group D at T3-T5 were increased and GABA was decreased, but without statistic significance. At T6 , the values recovered to the level at T0 . Conclusion Dexmedetomidine can be used to maintain hemodynamic stability in intracranial tumor surgery,and may play a role in cerebral protection through inhibiting expression of Glu and Asp (EAA).

OBJECTIVETo investigate the factors associated with insulin resistance (IR) in patients with chronic hepatitis B virus (HBV) infection.

METHODSSixty-eight patients with mild chronic hepatitis B (MCHB) caused by HBV were recruited for study. Sixty-seven healthy individuals with no hepatitis virus infections and normal liver function were enrolled as controls. Demographic, anthropometric, clinical, and blood biochemical parameters were compared between the two groups. IR was determined by the homeostasis model assessment (HOMA-IR). The MCHB group was further divided into patients with IR (HOMA-IR: > 2.7) and patients without IR (HOMA-IR: less than 2.7). Demographic, anthropometric, clinical, and blood biochemical parameters were compared between the two sub-groups. Finally, the potential factors associated with IR were evaluated.

RESULTSCompared to the healthy controls, the MCHB patients had significantly higher serum insulin (Z = -5.451, P less than 0.01), alanine aminotransferase (ALT) (Z = -8.211, P less than 0.01) and HOMA-IR (Z = -5.631, P less than 0.01). IR was detected in 44.12% (30/68) of the MCHB patients. The levels of ALT and body mass index (BMI) were significantly different between the MCHB patients with IR and without IR (t = -2.358, and t = -3.566, P less than 0.05). There was a significant correlation between BMI, ALT, and HOMA-IR in the MCHB patients (r = 0.374, r = 0.282, P less than 0.05), but not with the HBV DNA loads (r = 0.015, P = 0.904). Binary logistic regression analysis indicated that BMI [Exp(B): 1.859, P less than 0.01] and ALT [Exp(B): 1.022, P less than 0.05] were independent risk factors of IR in MCHB.

CONCLUSIONThere is a high prevalence of insulin resistance in patients with mild hepatitis caused by chronic HBV infection. In these patients, IR is correlated with abnormal liver function and BMI, and not HBV load.

Adult ; Alanine Transaminase ; analysis ; Blood Glucose ; Body Mass Index ; Case-Control Studies ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; metabolism ; virology ; Humans ; Insulin ; blood ; Insulin Resistance ; Male ; Middle Aged ; Viral Load

OBJECTIVE: To explore the feasibility and the value of Bayes analysis in clinical decision-making for solitary pulmonary nodules (SPNs). METHODS: We collected 352 consecutive SPN patients (malignancy, n=135; benignity, n=217) retrospectively to form the training set. Utilizing Bayes analysis, the prior odds of malignant SPNs and the likelihood ratios of clinical and CT findings were derived from the training set, which were then used to calculate the probability of malignancy in each SPN. Bayes analysis was also tested prospectively for its diagnostic validation and precision of predictive probability on the test set of 132 SPN patients (malignancy, n=61; benignity, n=71), and compared with the performance of physicians using routine judgment. The actual results of patients diagnosis were analyzed according to the scale of calculated malignant probability in SPNs. RESULTS: The sensitivity, specificity, and accuracy of Bayes analysis for the training samples were 88.9%, 93.1%, and 91.5%, respectively. In the test set, the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of Bayes analysis were 88.5%, 85.9%, 87.1%, 84.4%, and 89.7%, respectively. The accuracy of Bayes analysis had no statistically significant difference with that of senior physician A (80.3%, chi2=2.37, P=0.122) and B (79.5%, chi2=3.12, P=0.076), and was higher than that of junior physician C (74.2%, chi2=7.05, P=0.012) and D (74.2%, chi2=6.56, P=0.009); The Brier score was 0.099, 0.140, 0.137,0.154, and 0.179 for Bayes analysis,senior physician A, senior physician B, junior physician C, and junior physician D, respectively. Excluding the solitary metastasis (n=11) misclassified, the false negative rate of Bayes analysis was 1.0% (5/484) for SPNs with <20% estimated probability of malignancy. CONCLUSION Bayes analysis is accurate in qualitative diagnosis, precise in forecasting the malignant probability, and has low false negative rate for SPNs. It is feasible to use Bayes analysis for the management of SPNs.
Adenocarcinoma ; diagnosis ; Adult ; Aged ; Aged, 80 and over ; Bayes Theorem ; Decision Making ; Diagnosis, Differential ; Feasibility Studies ; Female ; Humans ; Lung Neoplasms ; diagnosis ; Male ; Middle Aged ; Solitary Pulmonary Nodule ; diagnosis ; Tomography, X-Ray Computed ; Tuberculoma ; diagnosis ; Tuberculosis, Pulmonary ; diagnosis

OBJECTIVETo construct the recombinant adenovirus expression vector of a short hairpin RNA (shRNA) targeting phosphatase and tensin homolog deleted on chromosome ten (PTEN) gene for gene therapy of ischemic cerebral injury.

METHODSThe U6 expression promoter and shRNA of pGenesil-1-shRNA, which was constructed and identified in our previous experiment, were subcloned to pAdTrack shuttle plasmid. The product pAdTrack-U6-shRNA was linearized by PmeI for homologous recombination with pAdEasy-1 in pAdEasy-1 competence bacteria. The positive clone was identified by enzyme digestion, PCR analysis and DNA sequence analysis. After linearization by PacI, the recombinant adenovirus DNA shuttle plasmid pAdEasy-U6-shRNA was transfected into 293 cells for packaging and amplification of Ad-U6-shRNA, which was further identified by PCR analysis and DNA sequence analysis. Western blotting was used to detect the expression of PTEN protein in the hippocampal neurons infected with the adenovirus.

RESULTSThe pAdTrack-U6-shRNA and pAd-U6-shRNA plasmids had been successfully constructed as verified by PCR analysis, enzyme digestion and DNA sequence analysis. PCR analysis and DNA sequence analysis confirmed successful packaging of the recombinant adenovirus Ad-U6-shRNA in 293 cells. PTEN protein expression decreased significantly in the hippocampal neurons after infection by the recombinant virus.

CONCLUSIONWe have successfully constructed the recombinant adenovirus Ad-U6-shRNA targeting PTEN gene, which provides a basis for investigating the role of PTEN in neuroprotection after cerebral ischemic injury using RNA interference.

Adenoviridae ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA

To investigate the effect of HBV on the expression of Sterol regulatory element binding proteins( SREBP ) in the hepatocyte of patients with chronic hepatitis B (CHB) combined with hepatic fatty change. 55 cases diagnosed as CHB combined with hepatic fatty change in our department were selected and liver biopsies were carried out. The patients were dividied into 3 groups, group A: HBV DNA is less than or equal to 1000 copies/ml(15 cases), group B: 1000 copies/ml less than HBV DNA less than 100000 copies/ml (18 cases) and group C: HBV DNA is more than or equal to 100000 copies/ml (22 cases). 10 patients with HBV DNA in less than or equal to 1000 copies/ml after antiviral therapy with Nucleoside analogues were seen as group C1 (before treatment) and group C2 (after treatment) respectively; 12 patients with HBV DNA is more than or equal to 100000 copies/ml after antiviral therapy were classified as group C3 (before treatment) and group C4 (after treatment). Lipid droplets in the hepatic tissue were observed with oil red staining. Real time PCR were performed to detect the expressions of SREBP-1c and SREBP-2 mRNA in the liver. The protein expressions of SREBP-1c and SREBP-2 were detected with immunohistochemistry staining. Statistic data were analysed with SPSS11.5 software. (1) Red integrated optical densities (IOD) reflected by lipid drops in group A, B and C are 1004.27+/-218.63, 1937.01+/-401.47 and 4133.79+/-389.28 respectively, the degree of oil red O in each group was different (F = 385.69, P is less than to 0.01), which is increased as HBV DNA load increasing; Red IOD in group C1, C2 and C3, C4 are 4020.84+/-326.64, 1012.02+/-244.89, 4189.18+/-329.21 and 4121.76+/-304.09 respectively. Compared with group C1, the degree of oil red O in group C2 is decreased and the difference is statistically significant (t = 22.55, P is less than to 0.01); However, the difference of the degree of oil red O between group C4 and C3 is not statistically significant. (2) Compared with group A, the expressions of SREBP-1c mRNA in group B and C are raised by 1.218+/-0.130 and 1.798+/-0.118 times respectively, among group A, B, C, the expressions of SREBP-1c mRNA are statistically significant different ( F = 297.47, P is less than to 0.01). The expressions of SREBP-2 mRNA in group B and C are decreased by 0.956+/-0.118 and 0.972+/-0.153 times as compared to group A. However, the difference of SREBP-2 mRNA expression among the 3 groups is not statistically significant ( F = 0.568, P is more than to 0.05). Compared with group C1, SREBP-1c mRNA in group C2 is decreased by 0.714+/-0.081 folds (t=11.224, P is less than to 0.01), while SREBP-2 mRNA in group C2 is raised by1.034+/-0.155 times(t=0.692, P is more than to 0.05). SREBP-1c mRNA and SREBP-2 mRNA in group C4 are raised by 1.012+/-0.206 times and decreased by 0.998+/-0.183 times as compared to group C3 without difference found (t=0.196 or 0.031, P is more than to 0.05). (3) the expressions of SREBP-1c protein in group A, B and C are 36257.21+/-5709.79, 50413.47+/-4989.28 and 71025.83+/-6047.13 respectively, and the difference is statistically significant among the 3 groups (F = 178.26, P is less than to 0.01); the expressions of SREBP-2 protein in group A, B and C are 32913.52+/-3951.21, 32625.91+/-4025.06 and 34173.44+/-5316.25 respectively, but the difference is not statistically significant among the 3 groups ( F = 0.562, P is more than to 0.05), SREBP-1c protein levels in group C1, C2, C3, C4 are 69832.16+/-4941.36, 48735.47+/-5471.41, 70871.69+/-5083.14 and 68913.32+/-5343.22 respectively, the difference of SREBP-1c protein levels between group C1 and C2 is statistically significant (t=10.260, P is less than to 0.01); while the difference between group C3 and group C4 is not statistically significant(t=1.558, P is more than to 0.05). The expressions of SREBP-2 protein in group C1, C2, C3 and C4 are 33 980.21+/-4081.80, 34011.50+/-3859.27, 33610.12+/-4761.10 and 32915.66+/-5023.61 respectively, the difference of SREBP-2 protein levels in group C1 and group C2 is not statistically significant (t=0.038, P is more than to 0.05) and same result exists between group C3 and group C4 (t=0.459, P is more than to 0.05). HBV DNA may participate in the hepatic steatosis formation through interfering with the SREBP-1c expression.
Fatty Liver ; metabolism ; Hepatitis B, Chronic ; metabolism ; Hepatocytes ; metabolism ; Humans ; Sterol Regulatory Element Binding Protein 1 ; metabolism

Objective To study the effect of operating rooms of two different cleanliness on surgical site infection (SSI)in neurosurgical craniocerebral operation,provide reference for the prevention and control of SSI.Methods Neurosurgical craniocerebral operation undertaking in grade I and II operating rooms in a hospital between July 2015 and July 2016 was performed retrospective study,patients in grade I operating rooms were as control group and those in grade II operating rooms were as trial group,occurrence of SSI was compared between two groups of pa-tients.Results A total of 454 patients undergoing craniocerebral operation were investigated,212 patients were in control group (grade I operating rooms)and 242 in trial group (grade II operating rooms).Patients in control group had 25 cases of SSI,incidence of SSI was 11.79%;trial group had 31 cases of SSI,incidence of SSI was 12.81%. Nine cases of SSI were superficial incision infection,there was no deep incision infection,the other 47 cases of SSI were intracranial infection(organ space infection).There was no significant difference in the incidence of SSI be-tween two groups of patients(P＞0.05).Conclusion Grade I and II cleanliness operating rooms have no direct effect on SSI in patients undergoing neurosurgical craniocerebral operation,comprehensive prevention and control mea-sures should be taken to prevent the occurrence of SSI.

Objective To observe the expressions ofinterleukin-17A (IL-17A) and transforming growth factor β (TGF-β) in the peripheral brain tissue of nidus after microsurgical resection of brain arterivenous malformations (AVM),and their significance.Methods We prospectively chose 25 patients with AVM; 12 of them were performed endovascular embolization and then microsurgical removal (Group A),and the other 13 were performed microsurgical removal only (Group B).Another 6 patients with traumatic brain injury and hypertensive cerebral hemorrhage that needed internal decompression were used as controls.The expressions of IL-17A and TGF-β in these brain tissues were detected by immunohistochemistry and RT-PCR.Results The protein and mRNA expressions of IL-17A in Group A and B were significantly increased as compared with those in the controls (P＜0.05);the protein and mRNA expressions of IL-17A in Group A were significantly decreased as compared with those in Group B (P＜0.05).The protein and mRNA expressions of TGF-β in Group A and B were significantly decreased as compared with those in the controls (P＜0.05); the protein expression of IL-17A in Group A was not significantly increased as compared with that in Group B (P＞0.05); the mRNA expression of IL-17A in Group A was significantly increased as compared with that in Group B (P＜ 0.05).Conclusion The increased expression of IL-17A might be associated with the growth and pathologic process of AVM; the increased expression of TGF-β might inhibit the inflammatory reaction after embolization.

OBJECTIVETo investigate the social mental state of drug addicts in a compulsive drug abuse treatment center; evaluate the effectiveness of integrated program for the prevention of abuse relapse and improvement of drug addicts' psychological health.

METHODSThe study subjects were addicts from the Wuhan Compulsive Drug Abuse Treatment Center between October 2003 and June 2004, who satisfied the inclusion criteria. A non-randomized control-intervention study design was adopted. Volunteers willing to take part in intervention were put into the intervention group with their full awareness and willingness to prevent drug abuse relapse. The control group was composed of the addicts who were willing to prevent relapse and to be followed up after their discharge.

RESULTSThe effectiveness of the integrated intervention program in promoting addicts' psychological health: before the intervention, the scores of Self-Rating Anxiety Scale (SAS), the positive and negative dimensionalities of Simple Coping Style Questionnaire (SCSQ) and Chinese Perceived Stress Scales (CPSS) had no significant differences between the intervention group and the control group. After the intervention, except that the SCSQ's positive dimensionality in the intervention group was significantly higher than that in the control group, other indices in the intervention group were lower. Before and after the intervention, the psychological health level in both the groups was lower than that in the normal population; there were significant differences between addicts and normal subjects in regards with all of the indices above.

CONCLUSIONDrug abuse was associated closely with addicts' social mental factors. The integrated intervention program can alleviate anxiety and stress, reduce co-morbid mental disorders and effectively improve their coping style. In conclusion, the program can promote addicts' psychological health significantly.

Adaptation, Psychological ; Adult ; Case-Control Studies ; Female ; Humans ; Male ; Mental Health ; Program Evaluation ; Stress, Psychological ; Substance-Related Disorders ; psychology ; therapy