This study was aimed to observe different forms of β-amyloid peptide (Aβ) and insulin degrading enzyme (IDE) in the hippocampus and cortex in order to further explore the role of Aβ and IDE on spleen yin deficiency di-abetes-associated cognitive decline (DACD), and the effect of Zi-Bu Pi-Y in method. The rats were randomly divided into five groups, which were the blank control (Cont) group, diabetes (DM) group, spleen yin deficiency (pi) group, spleen yin deficiency diabetes (piDM) group and Zi-Bu Pi-Y in recipe (ZBPYR) group. Soluble and insoluble Aβ in the hippocampus and cortex of rats were extracted by gradient centrifugation, and then measured by ELISA. The ex-pression of IDE was observed by western blot. The results showed that the content of soluble and insoluble Aβ1-42 in the hippocampus and cortex of the DM group and piDM group were higher than the Cont group. The soluble and in-soluble Aβ1-42 content in the hippocampus and cortex of the ZBPYR group were reduced compared with the DM group and the piDM group. The soluble Aβ1-40 in the cortex of the DM group, pi group and piDM group were in-creased compared with the Cont group (P < 0.05). The soluble Aβ1-40 content of the ZBPYR group was decreased compared with the DM group and the piDM group (P < 0.05). The expression of IDE protein was decreased in the hippocampus of the DM group and the piDM group compared with the Cont group (P< 0.05), and the IDE protein level in the hippocampus of the ZBPYR group was increased compared with the DM group and the piDM group (P<0.05). The expression of IDE protein in the cortex of the DM group, pi group and piDM group was lower than the Cont group (P< 0.05). The IDE protein level in the cortex of the ZBPYR group was reduced compared to the DM group (P< 0.05). It was concluded that the increased Aβ1-42 in brain may be a major pathological change of DACD and spleen yin deficiency DACD. The decreased IDE expression may be one of the reasons to induce increasing of Aβ1-42 level. The Zi-Bu Pi-Y in method may decrease the Aβ1-42 content by upregulating IDE protein expression.

Objective To explore the effects of human neuromelanin (NM) and doparmine melanin (DAM) on ferritin mRNA expression under oxidative stress in dopaminergic neuronal cells and glial cells. Methods SK-N-SH and U373 were chosen as models of human neuronal and glial cells.Fenton reagent (FR) was used as a direct inducer of oxidative stress,and human neuromelanin (NM) or dopamine melanin (DAM) were added to the cultures at the same time. Iron chelator desferrioxamine (DFO) was added in some experiments parallel.The changes of ferfitin mRNA expression were detected by real-time quantitative PCR. Results Ferritin mRNA in SK-N-SH cells were significantly up-regulated with FR or FR+DAM treatment than untreated cells,and FR+DAM induced more Ferritin mRNA expression than FR+hNM did.These differences are significant (P＜0.05).With DFO,ferritin mRNA expressions in FR, NM,FR+NM, DAM treated SK-N-SH cells were decreased significantly comparing to DFO free cultures(P＜0.05).While in U373 cells,the expressions of Ferritin mRNA had no significant differences with or without FR,NM, DAM, FR+NM or FR+DAM treatment (P＜0.05).Furthermore,Ferritin mRNA expressions showed no significant differences in U373 cells with or without DFO treatment(P＜0.05). Conclusions Ferritin has a protective effect on cells,DAM could not used to produce PD model.

OBJECTIVETo investigate the dynamic correlation between pre-S1 antigen, pre-S2 antigen and HBV DNA in the serum of chronic hepatitis B (CHB) patients undergoing nucleoside analogue therapy.

METHODS12 CHB patients with transient virological response after lamivudine treatment, and 20 patients treated with adefovir for 5 years were recruited in this study. Serum samples were collected at four time points when HBV DNA fluctuated sharply during lamivudine treatment, and at 0, 8, 12, 28, 52, 104, 156, 208, 260 weeks following adefovir treatment. HBV DNA was quantified by real-time PCR, pre-S1 and pre-S2 antigens were detected by ELISA.

RESULTSThe titers of pre-S1 and pre-S2 antigens were not correlated with the HBV DNA level in the serum of lamivudine treated patients. Only in one case of the adfovir treated patients, the decrease of pre-S1 and pre-S2 antigens was in parallel with the decrease of HBV DNA. Linear regression analysis indicated that neither pre-S1 antigen nor pre-S2 antigen was correlated with HBV DNA in the serum of lamivudine or adfovir treated patients (P more than 0.05).

CONCLUSIONOur results indicate that the titers of pre-S1 and pre-S2 antigens are not correlated with the serum HBV DNA in CHB patients undergoing nucleoside analogue therapy. Neither pre-S1 nor pre-S2 is a good predictor for the outcome of nucleoside analogue treatment.

DNA, Viral ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Real-Time Polymerase Chain Reaction

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis ; complications ; pathology ; Liver Neoplasms ; etiology ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate whether down-regulation of Twist1 could change sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol.

METHODSHNE1 cells were transfected with the small interfering RNA (siRNA) expression vector pSuppressor-Retro-Si-Twist, containing the short hairpin RNA (shRNA) sequence targeting the Twist gene-coding region by Fugene 6. Positive clones were then selected in Neomycin (400 microg/ml) for 21 days. The low expressions of Twist1 were examined by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blot. Drug sensitivity of si-Twist1 HNE1 to taxol was determined by Annexin V-fluorescein isothiocyanate( FITC)/propidium lodide (PI) double-labeled flow cytometry and detection of DNA ladder. The Effect of Twist1 inactivation on HNE1 cell proliferation was observed by MTT assay and flow cytometry.

RESULTSAnnexin V- FITC-PI assay showed that apoptosis ratio was 40.2% in si-Twist HNE1 after treated with 10 ng/ml taxol, significantly higher than that in the control siRNA group 24.3%. The deference had statistic meaning. After the re-expression of HNE1, apoptosis ratio was 44.80% +/- 4.80% (x +/- s) in low Twist1 protein expression group and that was 27.00% +/- 2.91% in high expression group. The deference had statistic meaning (t = 4.374, P = 0.049). Real time PCR test revealed apoptosis protein bcl-2 expression in si-Twist HNE1 was 0.28 +/- 0.05, significantly lower than that in the control siRNA HNE1 (0.57 +/- 0.08, t = 6.710, P = 0.021), nevertheless, significant bax and bcl-XL changes were not observed (t = 2.000, P = 0.184 and t = 1.502, P = 0.272). MTT and FCM showed that down-regulation of Twist1 did not alter cell proliferation rate (P>0.05).

CONCLUSIONSDown-regulation of Twist1 could increase drug sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol by inducing apoptosis. These results suggested that Twist1 may be a promising treatment target for nasopharyngeal carcinoma therapy.

Apoptosis ; drug effects ; Cell Line, Tumor ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; Nuclear Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA Interference ; RNA, Small Interfering ; Twist-Related Protein 1 ; genetics

Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA, Viral ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Adenoviridae ; isolation & purification ; Adenovirus Infections, Human ; diagnosis ; Female ; Humans ; Infant ; Male ; Pneumonia, Viral ; diagnosis

Objective To explore the protective effect of gallic acid in Phyllanthus emblica on high glucose－induced apoptosis of pancreatic islet β cells, and to provide a reference for the discovery of natural compounds for the treatment of diabetes. Methods In vivo experimental model, wistar male rats were used as in vivo subjects and 50 mg/kg STZ was injected intraperitoneally. After the model was successfully established, 25 mg/kg of gallic acid was given orally, and the positive drug was Sitagliptin. After 4 weeks of administration, the blood was taken and the pancreas was removed for HE staining. Western blot was used to measure the expression of NLRP3 and TXNIP in pancreatic tissue in high sugar state. In vitro model, insulinoma cell line INS－1 cells were used as in vitro targets to establish high levels. In sugar－induced apoptosis model, INS－1 cells were cultured in glucose－free RPMI 1640 complete medium supplemented with 25 mmol/L glucose. Gallic acid was used as the test sample. Experiments were divided into normal controls, high－sugar models, and low, medium and high levels of gallic acid groups. The cell viability was measured by MTT assay. The mRNA expression of NLRP3 and TXNIP in INS－1 cells was detected by QPCR and Western blot, and the expression of NLRP3 and TXNIP protein was detected.Results (1) INS－1 cells were cultured in a medium with glucose concentration of 25mmol/L for 48h, and the apoptosis rate was increased compared with the control group (P＜0.01), indicating that the apoptosis model was established successfully under high glucose conditions. (2) 10, 5, and 2.5 μmol/L GA were used to treat the control group and the high glucose model group cells respectively. The survival rate of the control group did not change significantly (P＞0.05) . Compared with the control group, the expression of NLRP3 and TXNIP in INS－1 cells in the high glucose model group was significantly different (P＜0.05);the protein expression level was significantly downregulated after GA treatment, and there was a statistical difference (P＜0.05) . Compared with the control group, the expression of NLRP3 protein in INS－1 cells in the high glucose model group was statistically different (P＜0.01), and the protein expression level was significantly downregulated after GA treatment (P＜0.01) ; The protein expression level was up－regulated (P＜0.05);the protein expression level after GA treatment was significantly down－regulated (P＜0.05); (4) The expression of NLRP3 and TXNIP mRNA in INS－1 cells was increased in the high glucose model group compared with the control group (P＜0.01) ; The expression of protein was significantly down－regulated after GA treatment (P＜0.01) . Conclusion The cells were cultured for 48 h in glucose－free RPMI 1640 complete medium supplemented with 25 mmol/L glucose. GA has no effect on the proliferation of normal INS－1 cells. GA protects INS－1 cells from apoptosis under high glucose conditions. The mechanism may be related to GA down－regulation of NLRP3 and TXNIP gene expression.

Objective To investigate the correlation between diabetes distress and social support of elderly patients with diabetes mellitus. Methods Three instruments,including the general information questionnaire,the diabetes distress scale (DDS)and the social support rating scale (SSRS)were used to investigate the correlation between diabetes distress and social support of 95 elderly patients with diabetes mellitus from the endocrinology department of two tertiary level first-class comprehensive hospitals from at Novenber 2017.Results The total scores of diabetes distress are (32.7±11.5),at the level of mild distress.The proportion of moderate/severe level is 31.6%(30/95).The diamensions of "routine life distress" and "emotional distress" are most prominent;Social support overall and all diamension scores are lower than the model (all P<0.05);There is a significantly negative correlation between social support and diabetes distress overall,physical distress,routine life distress and interpersonal distress (all P<0.05). Conclusions The elderly patients with diabetes mellitus have different levels of diabetes distress and low level of social support.There is a negative correlation between diabetes distress and social support.We should strengthen the evaluation of diabetes distress and social support of the elderly patients with diabetes mellitus in clinical work,and take individualized interventions to improve the level of social support,in order to decrease diabetes distress and improve the quality of life.

Objective: To collect, compare and analyze the changes in the strength and stability of the forearm Rou-Kneading manipulation before and after the training in Shaolin Neigong (internal Qigong). Methods: Ninety first-year undergraduates were randomized into three groups using the random number table method, with 30 people in each group. The Gongfa (Qigong method) group received training in both Shaolin Neigong and forearm Rou-Kneading manipulation. The manipulation group only received training in forearm Rou-Kneading manipulation. The control group only received training in forearm Rou-Kneading manipulation for one week. The ZTC-1 intelligent Tuina (Chinese therapeutic massage) manipulation parameter detection system was used to collect the wave crest, wave trough, and crest-trough difference of the strength and frequency of the forearm Rou-Kneading manipulation on the Z-axis (up and down), X-axis (left and right) and Y-axis (backward and forward) at weeks one, five and ten from the three groups. The collected data were then processed and analyzed. Results: The intra-group comparisons showed statistical significance in the Gongfa group and manipulation group (P<0.05). The strength and stability shown on the axes Z, X, and Y constantly grew with the increase of training time in the Gongfa group. The wave crest on the axes Z and Y steadily rose in the manipulation group, as well as the frequency on the axes Z, X and Y. The control group failed to show statistical significance in any of the three times of intra-group comparisons (P>0.05). The between-group comparisons showed statistical significance among the three groups at weeks five and ten (P<0.05). At week five, the wave crest on the axes Z, X, and Y, and the crest-trough difference on the axes X and Y were more prominent in the Gongfa group than in the manipulation group, showing statistical significance (P<0.05). At week ten, the wave crest, wave trough, and crest-trough differences on the axes Z, X, and Y were more prominent in the Gongfa group than in the manipulation group, presenting statistical significance (P<0.05). Conclusion: Practicing Shaolin Neigong can help the trainees reach the level of strength and frequency of Tuina clinicians in a shorter time in the forearm Rou-Kneading manipulation training. It can advance the efficiency in studying the forearm Rou-Kneading manipulation and promote the quality of the manipulation.