OBJECTIVE:To obverse the efficacy and safety of Liuwei dihuang pill combined with TCM prescription acupoint application in the treatment of type 2 diabetic constipation. METHODS:120 patients with type 2 diabetic constipation were random-ly divided into control group(60 cases)and observation group(60 cases). All patients received controlling blood pressure and regu-lating blood lipids drugs,diet control combined with exercise,and fixed dose of Metformin hydrochloride tablet(0.5 g/times,2 times a day)and Acarbose tablet(100 mg/times,3 times a day);based on it,control group received 5 mg Mosapride citrate tablet, 3 times a day;observation group additionally received 8 Liuwei dihuang pills(3 times a day)combined with TCM prescription acu-point application (in double Tianshu Point,double Yongquan point,double Shenque Point and double Zusanli). 7 d was regarded as a treatment course,and it lasted 3 courses. Clinical efficacy,blood glucose [2 h postprandial glucose(2 h PG),blood glucose (FPG)],the clinical symptom indexes(defecation interval time,each defecation time,weekly defecation frequency)before and af-ter treatment,and the incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than in control group,and the difference was statistically significant (P<0.05). Before treatment, there were no significant differences in the 2 h PG,FPG,defecation interval time,each defecation time and weekly defecation fre-quency between 2 groups(P>0.05). After treatment,2 h PG and FPG in 2 groups were significantly lower than before,and obser-vation group was lower than control group;defecation interval time and each defecation time were significantly shorter than before, and observation group was shorter than control group;weekly defecation frequency was significantly higher than before,and obser-vation group was higher than control group,The differences were statistically significant (P<0.05). During treatment,6 patients showed adverse reactions in control group,and 5 in observation group,and there was no significant difference in the incidence of adverse reactions between 2 groups(P>0.05). CONCLUSIONS:Liuwei dihuang pill combined with TCM priscription acupoint ap-plication shows good efficacy in the treatment of type 2 diabetic constipation. It can effectively improve constipation,reduce defeca-tion interval time and each defecation time,increase weekly defecation frequency and have no influence on blood glucose control with small adverse reactions and high safety.

OBJECTIVEExpress and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.

METHODSThe expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.

RESULTSSuitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.

CONCLUSIONSWe have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.

DNA, Single-Stranded ; genetics ; metabolism ; DNA-Binding Proteins ; chemistry ; isolation & purification ; metabolism ; Hepacivirus ; Hepatitis C ; metabolism ; virology ; Humans ; Molecular Weight ; Protein Binding ; RNA, Viral ; genetics ; metabolism

Objective To study the effects of shRNA-CtBP2 on the growth of prostate cancer PC3 cells. Methods There were three experimental groups in this study,which include blank control group, empty plasmid transfected group and transfected shRNA group. CtBP2 mRNA sequence is targeted by 3 pairs of designed interfering shRNA to built shRNA-Ct-BP2 recombinant plasmid then it is transfected into PC3 cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot assays were used to detect the transcription and expression levels of CtBP2 mRNA and protein, respective-ly. PC3l proliferation was measured by MTT assay. Results Builting shRNA-CtBP2 recombinant plasmid and transfect-ing PC3 cells were successful. Transcription and expression levels of CtBP2 mRNA and protein were significantly decreased in shRNA-CtBP2 transfected PC3 cells. After CtBP2 silencing, cell proliferation was blocked in the shRNA-CtBP2 cells compared to that of blank control group (P<0.01). Conclusion shRNA-CtBP2 could significantly inhibit CtBP2 expres-sion, suppress the growth of PC3 cells, which suggests that CtBP2 may be a new target for PCa gene therapy.

Objective To study the expression of monocyte chemotactic factor -1(MCP-1) and fibronectin (FN ) in secondary acquired middle ear cholesteatoma epithelium ,and to investigate the ability of cholesteatoma of e-rosion .Methods MaxVisionTM immunohistochemical method was used to detect the expression of MCP -1 and FN in the secondary acquired middle ear cholesteatoma tissues from 30 patients ,in the retroauricular skin from 20 pa-tients and in the retroauricular skin from 16 normal subjects .Then we scanned it into a computer by an image scan-ner and quantified the gray value of them using commercial software .Results MCP-1 appeared to be localized in all epithelial layers of middle ear cholesteatoma ,particularly in the spinous layers .The positive expression rates of MCP-1 was 70% ,the gray value was 147 .2 ± 20 .1 ,which were siginificantly higher than those of in the retroau-ricular skin from patients(35% ,200 .8 ± 18 .4)and from normal subjects(37 .5% ,193 .3 ± 15 .5)(P<0 .05) .The ex-pression of FN in all epithelial layers of middle ear cholesteatoma were abundantly stained ,especially in the basal and spinous layers and the matrix of cholesteatoma .The positive expression rates of FN was 76 .7% ,the gray value was 147 .2 20 .1 ,which were siginificantly higher than those of in the retroauricular skin from patients (30% ,195 .0 ± 12 .9)and from normal subjects(31 .3% ,191 .6 ± 13 .5)(P<0 .05) .It showed statistically significant correlation between the expression of MCP -1 and FN and the erosion ability of middle ear cholesteatoma (rmcp-1 = -0 .682 , rfn = -0 .531 ,P<0 .01) .There was not linear correlation between the expression of MCP -1 and FN .Conclusion MCP-1 and FN are overexpressed in middle ear cholesteatoma .There was correlation between the expression of MCP-1 or FN and the erosion ability of middle ear cholesteatoma ,indicating that MCP -1 and FN may play an im-portant roles in invasive behavior of cholesteatoma .

The clinical data of 459 patients,who were first diagnosed as HIV/HBV co-infection from January 2007 to December 2013,were retrospectively analyzed.Among all patients,there were 89 cases with CD4 ＜ 50/μl,134 cases with CD4 50-200/μl and 236 cases with CD4 ＞ 200/μl,when HIV infection was diagnosed.In these three groups with different CD4 levels,the HBV DNA positive rates were 49.3％ (37/75),50.5％ (54/107) and 33.7％ (66/196);the HBV viral load were (6.37 ± 1.71) log10 copies/ml,(5.82 ± 1.86) log10 copies/ml and (4.36 ± 1.64) log10 copies/ml;the rates of abnormal liver function were 29.2％ (26/89),29.1％ (39/134) and 10.6％ (25/236);the occurrence rates of end-stage-liver-diseases were 16.9％ (15/89),14.9％ (20/134) and 5.1％ (12/236);the mortality rates were 10.1％ (9/89),9.7％ (13/134) and 3.8％ (9/236),respectively.The HBV DNA positive rates,HBV viral load,the rates of abnormal liver function,the occurrence rates of end-stage-liver-diseases and the mortality rates in CD4 ＞ 200/μl group were lower than that in CD4 ＜ 50/μl group and 50-200/μl group.The results suggest that for HIV and HBV co-infection patients,HBV replication level and prognosis of liver diseases are associated with CD4 + T lymphocyte count.

Melanin is formed by oxidative polymerization of phenolic compounds,basically different kinds of melanin come from different organisms.DOPA melanin and DHN melanin have same physical and chemical characters although they have different biosynthetic pathway.DHN melanin is common in plant fungi and plays an important role on infection.The melanin accumulates in the fungal cell walls and prevents organic and inorganic molecules penetrating out,that insures appressorium's pressure and infection ability.This paper has reviewed the kinds and characters,especially discussed the role of melanin during pathogen infection based on our some research.

?AlM: To evaluate the long - term effects of laser peripheral iridectomy ( LPl ) and trabeculectomy in treating early chronic angle-closure glaucoma.
?METHODS: Ninety-eight patients (102 eyes) with early chronic primary angle-closure glaucoma were randomly divided into two groups. Group A of 50 patients (54 eyes) was treated with LPl and group B of 48 patients (48 eyes) with trabeculectomy. After 3 - 8y of follow - up observation, comparison would be made from the perspectives of postoperative eyesight, intraocular pressure, anterior chamber angle, visual field and cup/disc ratio ( C/D) .
?RESULTS:ln group A, 24 eyes with eyesight declining, 22 eyes with theintraocular pressure>21mmHg (1mmHg=0. 133kPa), 21 eyes with chamber angle synechia >180o, 21 eyes with visual field narrowed, 21 eyes with C/D ratio enlarged. The results of group B for the same items were 10, 5, 4, 4, 4 eyes respectively. The comparative difference was statistically significant (P<0. 05).
?CONCLUSlON:Good effects will be achieved for early-stage chronic angle - closure glaucoma with surgical method. Trabeculectomy is obviously better than LPl for the long-term effects.

A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

AIM:To observe related biological parameters of 3 minutes dark-room provocative test in patients with laser peripheral iridectomy(LPI) in the fellow eyes of acute primary angle-closure (APAC) by ultrasound biomicroscopy (UBM).To explore the risk factors in primary angle closure suspect(PACS) patients with progressive angle closure after LPI.METHODS: Seventy-eight eyes of APAC patients without peripheral anterior synechia were selected.Each eye underwent 3 minutes dark-room provocative test after LPI.Anterior segment parameters, including anterior chamber depth (ACD), anterior chamber angle open distance500 (AOD500), peripheral iris thickness (PIT), iris convex (IC), the position of iris insertion and trabecular-ciliary process distance (TCPD), and the number of positional angle closure(NPAC) were observed and analyzed by statistic methods.RESULTS:Patients with APAC were examined by UBM after LPI and 26 eyes(33%) occurs at least one positional angle closure,19 eyes(24%)were positive in 3 minutes dark-room provocative test among them.It occurs a positive relationship between the elevation intraocular pressure and the number of positional angle closure in dark-room provocative test(r=0.84, P<0.01).AOD500, IT and IC were significantly changed from normal light to darkroom between positional angle closure positive group and positional angle closure negative group(all P<0.01).In single factor analysis, AOD500(P=0.003), IT(P=0.012), IC(P=0.043), TPCD(P=0.015), the position of iris insertion(P=0.024) were correlative factors of positive results.In multiple-factor analysis, only IT(P=0.011), TPCD(P=0.009), iris root attachment points(P=0.02) were independent risk factors of positive results.CONCLUSION:A certain proportion of patients with PACS after LPI appeared positional angle closure in a dark room.Peripheral iris hypertrophy, anterior displacement of the ciliary body and iris root attachment points are vital risk factors.Long-term follow-up study and intervention treatment are required in these patients after LPI.