Diabetic microangiopathy is one of the most common chronic diabetic complications,which is associated with the genetic predispositions and environmental factors.Well-controlled blood glucose alone can not reduce the morbility or prevent the process of diabetic microangiopathy.The therapy aiming at the pathogenesis is an urgent request.The various proteins related to the development and progression of diabetic microangiopathiy were reviewed,aiming at providing the potential target for early diagnosis and efficient therapy.

Objective To use gene engineering technique to express the humanin.Methods Humanin segment was synthesized and cloned into the prokaryocyte expression vector pGEMEX-1 and pet44a. The plasmid was transformed to E. coli BL21(DE3) by screening and identifying to obtain the high expressing positive clone.Results The humanin segment was cloned into vector successfully,and expressed efficiently with the inducement of IPTG. The molecular weight of the expressed recombinant protein of pGEMEX-1-humanin was about 28 kd while it was 63 kd in pet44a-humanin,and the expressed quantity of fusion protein was 40 percentage of the total bacterial protein while it was 20 percent in pet44a-humanin.Conclusion The humanin has been cloned and expressed successfully and the high efficiently expressed recombinant protein might contain the intact important structure domain of the humanin.

Objective To investigate the efficacy and safety of the combined therapy of cyclophosphamide and thalidomide in the treatment of refractory Crohn's disease (CD).Methods This study was a prospective and open study.A total of 15 patients with refractory CD were enrolled.All patients received intravenous cyclophosphamide 200 mg every other day for two weeks,then followed by intravenous 400 mg once a week until the cumulative dose reached 6 to 8 g.when the cyclophosphamide treatment started,at the same time thalidomide was taken 25 to 75 mg every night according to the tolerance of patients.Before the treatment,two weeks' after the treatment and at the time when the cumulative dose of cyclophosphamide reached 6 to 8 g,Crohn's disease activity index (CDAI),hemoglobin (Hb),white blood cell (WBC) count,erythrocyte sedimentation rate (ESR) and high-sensitivity C-reactive protein (hs-CRP) were recorded.Endoscopy examination was conducted before the treatment and at the time when the cumulative dose of cyclophosphamide reached 6 to 8 g.The condition of mucosa healing was observed and scored by simple endoscopic score for crohn's disease (SES-CD).Adverse effects of all patients were monitored.Paired t test was performed for statistical analysis.Results Before the treatment,the CDAI of 15 patients with refractory CD was 235.87±59.87,two weeks after the treatment the CDAI declined to 135.33 ± 29.23,and the difference was statistically significant (t=7.50,P＜0.01).Before the treatment,ESR and hs-CRP was (42.13±22.80) mm/1 h and (13.73± 2.18) mg/L.Two weeks after treatment they declined to (23.80±16.63) mm/1 h and (5.77±4.77) mg/L,and the differences were statistically significant (t=2.43 and 6.17,both P＜0.05).After two-week treatment,10 patients achieved clinical remission.After the cumulative dose of cyclophosphamide reached 6 to 8 g combined therapy,CDAI of patients was 108.14 ± 47.10,which decreased significantly compared with that before treatment (t=6.30,P＜0.01).ESR,hs-CRP and WBC count was (19.35± 19.18) mm/1 h,(6.16± 5.02) mg/L and (6.28 ± 3.42) × 109/L,respectively,which decreased compared with those before treatment,and the differences were statistically significant (t=5.90,5.40 and 3.71,all P＜0.01).Twelve patients achieved clinical remission.And the lesions of 12 patients improved under endoscope,furthermore,the mucosa of four patients healed.Before the treatment,SES-CD was 9.14 ± 5.39,which declined to 5.07 ± 4.58 after the treatment,and the difference was statistically significant (t =3.14,P ＜ 0.01).During the treatment,five patients had adverse effects.Alanine aminotransferases (ALT) increased in three patients,WBC count decreased in one patient and one patient got a severe urinary infection.Conclusions Patients with refractory CD could achieve clinical remission,mucosa healing under endoscopy and better efficacy with the combined therapy of cyclophosphamide and thalidomide.However,adverse effects should be monitored during the treatment.

AIM: To investigate the mechanism of the AP-1 signal transduction pathway inhibited by JIP in nasopharyngeal carcinoma cells. METHODS: AP-1 activity was triggered by Dox-induced LMP1 expression in Tet-on-LMP1-HNE 2 cells (L7). The retention of phospho-JNK in the cytoplasm caused by JIP was examined with immunofluroscence assay. RESULTS: 24 h after transfection of L7 cells with the JIP expression plasmid, the translocation of activated JNK was inhibited, which resulted in the retention of phospho-JNK in the cytoplasm and down-regulation of the AP-1 activity. CONCLUSION: JIP down-regulates the activity of AP-1 through the inhibition of the translocation of JNK.

OBJECTIVETo develop the knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for occupational groups, and to provide a convenient and effective tool for the survey of knowledge, attitude, and behavior on the prevention and control of occupational diseases in occupational groups and the evaluation of intervention effect.

METHODSThe initial questionnaire which was evaluated by the experts was used to carry out a pre-survey in Guangzhou, China. The survey results were statistically analyzed by t test, identification index method, correlation analysis, and Cronbach's a coefficient method. And then the questionnaire was further modified, and the content of the questionnaire was determined finally.

RESULTSAfter modification, there were 18 items on knowledge, 16 items on attitude, and 12 items on behavior in the "Knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for enterprise managers"; there were 19 items on knowledge, 10 items on attitude, and 11 items on behavior in the "Knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for workers".

CONCLUSIONThe knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for occupational groups is developed successfully, and it is a convenient and effective tool for the survey of knowledge, attitude, and behavior on the prevention and control of occupational diseases in occupational groups and the evaluation of intervention effect.

China ; Health Knowledge, Attitudes, Practice ; Humans ; Occupational Diseases ; prevention & control ; Surveys and Questionnaires

OBJECTIVETo seek effective drugs for anti-hepatitis C virus by screening 20 Chinese herbs often used for clearing heat and dissipating toxin with nude mice model of hepatitis C viral (HCV) infection.

METHODSAfter the model mice had been treated with selected drug for 3 months, transmission electron microscope was used to observe whether the HCV-like particles in human fetal hepatocytes (HFH) transplanted into mice spleen still existed, and quantitative RT-PRC technique was used to detect the serum content of HCV-RNA before and after treatment.

RESULTS(1)HCV-like particles existed in all the model mice after treatment. (2) Serum content of HCV-RNA decreased after treated with Radix Gentianae, Radix Scutellariae, Radix Sophorae tonkinensis, Fructus Gardeniae and Fructus Sophorae flavoscentis, but unchanged after treatment with other drugs.

CONCLUSIONAll the 20 herbs screened has not effect in directly eradicating HCV, but Radix Gentianae, Radix Scutellariae, Radix Sophorae tonkinensis, Fructus Gardeniae and Fructus Sophorae flavoscentis could significantly inhibit the replication of HCV-RNA.

Animals ; Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gentiana ; chemistry ; Hepacivirus ; drug effects ; Hepatitis C ; virology ; Male ; Mice ; Mice, Nude ; Phytotherapy ; Scutellaria baicalensis ; chemistry ; Sophora ; chemistry ; Virus Replication ; drug effects

OBJECTIVETo construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin.

METHODSThe recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing.

RESULTSRecombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence.

CONCLUSIONSA eukaryotic expression plasmid of Humanin was successfully constructed.

Base Sequence ; Cloning, Molecular ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; genetics ; Protein Engineering ; methods ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry

?ig-h_3 was first identified as a transforming growth factor-beta1-inducible gene in human lung adenocarcinoma cell line. It encodes for a secreted extracellular matrix (ECM) protein, which is thought to act on cell attachment and ECM composition. Previous study showed that ?ig-h_3 were highly expressed in human hepatoma cell lines and lowly expressed in human normal hepatic cells. The present study aimed to transfect ?ig-h_3 into 7721 cells to investigate its effect on secretion of MMPs in the transfected human hepatoma cells. Full-length ?ig-h_3 gene,cloned by reverse transcription polymerase chain reaction (RT-PCR) was inserted into the eukaryotic expression vector pEGFP-C_2. The recombinant plasmid was transfected into 7721 cells with Lipofectamine2000 and Gelatin-Zymography were adopted to detect the production of MMPs in the transfected cells. Results showed that ?ig-h_3/pEGFP-C_2 recombinant expression plasmid was successfully constructed and achieved high transfection efficiency. MMPs expression of the transfected cells was promoted significantly. These results suggest that overexpression of ?ig-h_3 promoted the production of MMPs, indicating that ?ig-h_3 may play roles in the invasive and metastatic processes of hepatoma.

The zinc chloride and cystine resistant strain of S.cerevisiae YZM-14(ZnCl2r,Cysr) was screened with the mutant processing of the protoplast of S.cerevisiae by combinative mutagens of ultraviolet and nitrite.The glutathione(GSH) production(84.72 mg/L),dry cell weight(7.63 g/L) and the intracellular GSH content(11.10 mg/g) of YZM-14 were 2.79,1.63 and 1.71 times compared with that of the initial strain.The biosynthetic process of GSH was divided into three phases according to the time course of the specific cell growth rate and GSH yielding coefficient.In the second phase,the metabolic flux of the pentose phosphate pathway and the GSH precursors biosynthetic pathway of the mutant strain increased by 8.1 mmol/(g?h),compared with that of the initial strain.Furthermore,the metabolic flux of the organic acids secretion of the mutant strain decreased.Through these mechanisms,the utilization efficiency of the carbon sources was enhanced and high production of GSH was obtained.

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.