OBJECTIVE:To develop a HPLC method for determination of the content of baicalin in Kugan gran?ules.METHODS:ODS-C 18 column was used and methanol-water-acetic acid(45∶55∶1)was used as mobile phase and UV detection wavelength was274nm.RESULTS:There was a good linear relationship between the concentration of Baicalin and absorption area value in range of0.25?g～2.5?g r=0.9992.The average recovery was98.4%with RSD=1.09%(n=5). CONCLUSION:The method was highly repeatable and accurate.

Objective:To investigate the inhibitory effect of Epibueropyridinium A on aldose reductase.Methods: Aldose reductase was extracted from cattle crystalline lens.Besides Epibueropyridinium A,the reactive system also contained DL-glyceraldehyde,aldose reductase,and NADPH.The activity changes of aldose reductase were detected at 340 nm.Epalrestat was taken as the positive control.The inhibitory type,Ki and IC_(50) were determined by double reciprocal plot,quadratic drawing,and drawing of inhibitor's concentration to inhibitory ratio,respectively.Results: Epibueropyridinium A significantly inhibited the activity of aldose reductase in a competitive manner,with IC_(50) being 4.2 ?g/ml and Ki being 4.88 ?g/ml.Conclusion: Epibueropyridinium A is competitive inhibitor of aldose reductase.

OBJECTIVE: To determine the protective effects of nourishing spleen yin recipe (Zibu Piyin Recipe, ZBPYR), a compound traditional Chinese herbal medicine, on endoplasmic reticulum (ER) in neuronal cells responding to the stress by using sero-pharmacological method. METHODS: The mouse neuroblastoma cell line Neuro2a cells were treated with tunicamycin (Tm, an inhibitor of N-glycosylation). The ZBPYR-treated cell group was established by incubating cells with ZBPYR serum for one hour and treated with Tm. Reverse transcriptase PCR (RT-PCR) was utilized to detect the mRNA expressions from two genes after treatments, ER molecular chaperone glucose regulated protein 78 (GRP78) and transcriptional factor CCAAT/ enhancer-binding protein-homologous protein (CHOP). Lactate dehydrogenase (LDH) assay was also carried out to determine the LDH leakage from Neuro2a cells after treated with Tm and staurosporine (STS). RESULTS: The ZBPYR-treated cell group at all tested ZBPYR dosages showed significantly reduced expressions of both genes compared with Tm (5 microg/ml) treated control group (P<0.05). Therefore, ZBPYR serum inhibited the expressions of GRP78 and CHOP in mRNA level under ER stress induced by Tm. Different concentrations of ZBPYR serum pretreatment reduced the LDH leakage compared with the Tm and STS groups (P<0.05). Therefore, ZBPYR serum may inhibit the LDH leakage induced by Tm and STS. CONCLUSION: ZBPYR has neuroprotective effects. The mechanisms may be associated with inhibition of ER stress and mitochondrial dysfunction.

Objective To investigate the role of cHSP60 in the pathogenesis of murine cervicitis.Methods Fifty female C3H/HeN mice were randomly and equally classified into 5 groups, including the control group receiving no treatment and 4 groups receiving intravaginal inoculation of cHSP60 (cHSP60 group),live elementary bodies of Chlamydia trachomatis mouse pneumonitis (MoPn group), inactive elementary bodies of MoPn (inactive MoPn group) and growth medium (medium group), respectively. Five days after the inoculation,cervical tissue was resected from these mice and subjected to pathological examination. Results There were varying degrees of inflammatory reaction characterized by neutrophil infiltration, necrosis and shedding of mucosal cells in the cervices of mice in cGSP60 and MoPn groups. No statistical difference was observed in the incidence of cervicitis (90% vs. 80%, P ＞ 0.05), neutrophile count [76.00 (25.0 - 80.0) vs. 25.00 (8.75 -32.5), P＞ 0.05] or inflammation score [12.5 (11.5 - 14.25) vs. 9.00 (8.00 - 11.5), P ＞ 0.05] between the cHSP60 and MoPn group. The inflammatory reaction was weak with decreased incidence of cervicitis (40%),inflammation score [0.00 (0.00- 12.50)] and neutrophile count [0.00 (0.00- 15.50)] in inactive MoPn group compared with the cHSP60 and MoPn groups (all P ＜ 0.05). A small number of neutrophils migrated into the superficial layer of cervical mucosa in only 2 mice in the medium group. Conclusion cHSP60 may be a primary pathogenic factor in chlamydial genital tract infection.

Objective To investigate the prevalence, clinical characteristics and antibiotic resistance of Haemophilus influenzae (HI) in children with acute respiratory tract infection in Suzhou. Methods Data of sputum culture of 3 167 hospitalized childhood patients with acute respiratory tract infection from January 2006 to December 2007 were collected. The incidence of positive HI and the rate of resistance to different antibiotics were calculated and beta-lactamases of the strains were detected. Results About 4.4% of total 3 167 eases were infected with HI. The infection rate was related with season and sex, more frequent between February and June, more common in boys than girls. Children younger than three years old were likely to be infected by HI, eompared with other age groups. The beta-lactamase positive rate of HI was 31.4%. The resistance rates to ampicillin, SMZ + TMP, chloramphenicol, cefaclor, ceftazidime, tetracycline and ampicillin/sulbactam were 29.6% ～ 31.9%, 66.2% -73.9%, 19.7% ～ 15.9%, 2.8% ～ 14.5%, 2.8% ～0、 28.2% ～ 2.9% and 4.2% ～ 1.4% respectively. Isolates resistance to cefuroxime、 ceftriaxone、 imipenem、azithromycin and ciprofloxacin were not found. Conclusions The infection of HI in children with actue respiratory tract infection is closely related with season and sex in Suzhou. Children younger than three years old are at high risk. The beta-lactamase positive rate of HI was high and increased rapidly. Resistance rate to azithromycin, SMZ + TMP and chloramphenicol was high, some isolates were resistant to the second, third generation of cephalosporin. Monitoring the antibiotic resistance of H! should be emphasized.

Objective To explore the expression of COX-2 and p38MAPK in patients with trauma MODS. Methods Forty MODS patients were evaluated. The levels of peripheral blood mononuclear cells COX-2 and p38MAPK in MODS patients and 40 normal controls was detected by enzyme linked immunosor-bent assay (ELISA). RT-PCR was used to measure the COX-2 mRNA and p38MAPK mRNA expression of in PBMCs. ANOV and correlation analysis were used in statistical analysis. Results The levels of COX-2 and p38MAPK of PBMCs and the mRNA expression in MODS group were higher than in control group (all P <0.05). The levels of COX-2 and p38MAPK of PBMCs and the mRNA expression in dead group were higher than in survival group( all P <0.05). The levels of COX-2 and p38MAPK of PBMCs were positively correlated, (r =0.6 147, P<0.01). The expression of COX-2 mRNA, p38MAPK mRNA of PBMCs and APACHE I scoring were positively correlated (r1 =0.5 009, P1 <0.05,r2 =0. 5 316, P2 <0. 05). Conclusions COX-2 and p38MAPK of PBMCs take part in the onset of MODS, and may service as index to judge the prognosis of MODS.

Objective:In order to research the relationship between the function and the structure of human brain Acetylcholinesterase,use the monocolonal antibodies scanning antigenic decapeptides of human brain Acetylcholinesterase.Methods:Synthesis of overlapping decapeptides corresponding to the sequence of the human brain Acetylcholinesterase has been carried out on biotinylated with primary amino groups according to a method developed by M Geyson. Peptides Synthesized using the multipincombinatorial chemical synthesis technique have been used in an enzyme linked immunoabsorbent assay.Results:A antigenic epitope of human brain Acetylcholinesterase recognized by monoclonal antibody against Torpedo Acetylcholinesterase is received by decapeptide scanning.Conclusion:The antigenic epitope 111 could be recognized by the polycolonal antibody against human brain Acetylcholinesterase and monoclonal antibody against Torpedo Acetylcholinesterase,the results indicate that the epitope is highly conserved in human brain AChE and Torpedo AChE.

Objective To evaluate the effects of dexmedetomidine on the expression of phosphor-cAMP response element binding protein (p-CREB) in isoloated hippocampal neurons of fetal rats.Methods SpragueDawley rats on 16-18 days of gestation were sacrificed and the fetal rats were obtained.The hippocampi of fetal rats were isolated and hippocampal neurons were seeded in culture medium for 8 days.The cells were then divided into 4 groups (n =12 each) using a random number table:control group (group C),dexmedetomidine 0.001 μmol/L group (group D1),dexmedetomidine 0.010 μmol/L group (group D2),and dexmedetomidine 0.100μmol/L group (group D3).In D1.3 groups,dexmedetomidine with the final concentrations of 0.001 μmol/L,0.010 μmol/L,and 0.100 μmol/L was added to the culture medium,respectively,and then the cells were incubated for 3.5 h.The apoptosis in hippocampal neurons was detected by flow cytometry.The expression of p-CREB in hippocampal neurons was determined by RT-PCR and Western blot.Results Compared with group C,apoptosis rate was significantly decreased and the expression of p-CREB was up-regulated in D1.3 groups.Conclusion Dexmedetomidine inhibits apoptosis in isolated hippocampal neurons of fetal rats by up-regulating the expression of p-CREB.

Objective To establish a rapid SNP( single-nucleotide polymorphism) genetic identification method for the frozen samples, such as frozen embryos and sperm of inbred mice.Methods In this study, the frozen embryos and sperm of inbred mice were provided by Shanghai Lab.Animal Research Center.Whole genome amplification and PCR-LDR genotyping system were used to get the rich DNA sample.Forty-five SNP were genotyped by multiple polymerase chain re-action and ligase detection reaction( PCR-LDR) .Results The electrophoresis results showed that the whole genome am-plification technique could highly increase the total DNA of frozen embryos.PCR-LDR typing method was suitable for the mouse genome typing of 45 SNPs.Ten strains of inbred frozen embryos and sperms of C57BL/6, BALB/c, FVB/NJ mice were genotyping identified, and their SNP loci data obtained by PCR-LDR were as the same as those of database.The num-ber of frozen mouse embryos was proportional to the number of SNPs detected, and when the embryo number reached more than 12, the detection rate of SNP was 100%.Conclusions This method can be used to the genetic quality identification, and rapidly identify the inbreed frozen mouse embryos and sperms.