Objective: To analyze the correlation between HPLC chromatogram and the effect of different extracted fractions from Angelicae Sinensis Radix (ASR) on reinforcing Qi. Methods: Qi deficiency model of mice was set up by controlling diet and fatigue. The influences of the different extracted fractions from ASR on phagocytotic ability of mononuclear macrophage and immune organs of mice with the symptoms of Qi deficiency were compared, and the active fraction of ASR was screened. The part which was extracted with 70% ethanol (ethanol extract) from ASR has the certain improvement on the Qi deficiency symptoms. The peak areas of each common peak from HPLC fingerprint were correlated with the pharmacodynamic efficacy, and the grey relation statistics was used to study the spectrum-effect relationship. Results: The contribution of various components in ethanol extract with the effect of reinforcing Qi was determined by the spectrum-effect relationship, the order was as follows (characteristic peak number): 18 > 5 > 7 > 12 > 17 > 4 > 19 > 2 > 20 > 6 > 8 > 3 > 14 > 1 > 16 > 11 > 9 > 13 > 15 > 10, peak 7, 10, 11, 13, 17, 18, and 20 are ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A, ligustilide, butylidenephthalide, and levistilide A, respectively. Conclusion: The ethanol extract of ASR has the strong activity of reinforcing Qi, and there is a certain correlation between HPLC specific chromatogram and the effect.

HPS1-D, an active polysaccharide,was isolated and purified from Hedysarum polybotrys. HPS1-D was obtained after treated with Savage method and H2O2, and purified with DEAE-cellulose 52 and Sephadex G-100 gel filtration chromatography. Then physicochemical property analysis, GC, methylation, partial acid hydrolysis, and NMR method were used to study chemical structural of HPS1-D. The conformation was primarily analyzed with GPC-MALLS method and Congo red reaction. The anti-complementary activity of HPS1-D was evaluated with the hemolysis assay. HPS1-D was a heteropolysaccharide and consisted of D-glucose, L-arabinose, (7.2:1.3). HPS1-D proved to be a neutral sugar, with 1, 4-and 1, 4, 6-alpha-D-glucopyranosyl residues in backbone ,and 1, 5-and 1, 3, 5-alpha-L-arabinofuranosyl residues in branches. HPS1-D has a random coil state conformation with monodisperse mass distribution in 0.9% NaCl solution. And HPS1-D had triple-helix conformation in concentrate of NaOH solution. Anti-complementary activity of HPS1-D was closed to its positive control heparin.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Hemolysis ; drug effects ; Mice ; Plant Extracts ; chemistry ; pharmacology ; Polysaccharides ; chemistry ; pharmacology

Objective:To investigate the relationship between primary resection and KRAS gene mutation in the mild symptomatic patients with stage Ⅳ colorectal cancer,and to clarify its significance of prognosis.Methods:The clinical data of 46 mild symptomatic patients with stage Ⅳ colorectal cancer 2010 to December 2010 were collected.All the patients received primary resection.The KRAS gene mutation in the patients was detected by direct sequencing and the patients were followed up for 5 years.The influence of primary resection and KRAS gene mutation in prognosis of the patients with stage Ⅳ colorectal cancer was analyzed, and the clinical pathological features which might influence the prognosis were analyzed by survival analysis.Results:In 46 patients with colorectal cancer, KRAS gene mutation was found in 20 cases, the mutation rate was 43.4%, and most mutation was found at Codon 12. The KRAS mutation had relationship with the tumor site and multiple metastasis (P<0.05).The Kaplan-Meier survival curve and univariate analysis results showed that the median survival time of the patients with wild type of KRAS gene was 58.4 months, the median survival time of the patients with mutant type of KRAS gene was 42.2 months, and there was no significant difference (P>0.05).The median survival time of right colon cancer patients was 34.2 months, the median survival time of left colon cancer patients was 58.3 months, and there was sigificant difference (P<0.05).The cancer metastases including liver, lung and multiple metastasis were closely related to the poor prognosis of the colorectal cancer patients(P<0.05).The median survival time of the patients with stage Ⅳ colorectal cancer was 39.6 months after operation.Conclusion:After primary resection of the mild symptomatic patients with stage Ⅳ colorectal cancer,the median survival time of the patients with colorectal cancer in left colon site and right colon site were prolonged.Right colon cancer has more poorer prognosis than left colon cancer.KRAS gene mutation is associated with the tumor site and the multiple metastasis.The location of metastasis affect the prognosis.

AIM:To investigate the clinical significance of microRNA-26a-5p (miR-26a-5p)-regulated mye-loid cell leukemia-1 (MCL-1) expression in the development of maternal preeclampsia.METHODS:Plasma and placen-tal tissues were collected from 21 cases of normal pregnancy, 13 cases of maternal gestational hypertension, 15 cases of mild preeclampsia and 26 cases of severe preeclampsia.The levels of plasma and placental miR-26a-5p and placental MCL-1 mRNA were detected by real-time PCR.Western blotting analysis was used to determine the protein expression of placen-tal MCL-1.The clinical significance of the above parameters was also analyzed.RESULTS:miR-26a-5p expression gradu-ally increased(P<0.01) in the 4 groups of maternal plasma and placentas with the disease development, and the mRNA expression of MCL-1 was significantly reduced in the placentas (P<0.01), both showing a significant negative correlation (P<0.01).Meanwhile, the expression of miR-26a-5p and MCL-1 protein in the placental tissues was negatively correla-ted (P<0.01).The miR-26a-5p up-regulation in maternal plasma and placental tissues was negatively correlated with ges-tational age, maternal plasma albumin levels and fetal weight, while it was positively correlated with maternal blood pres-sure and urinary protein level (P<0.01), which was in contrary to the down-regulation of placental MCL-1.CONCLU-SION:Up-regulation of miR-26a-5p is involved in the occurrence and development of preeclampsia by down-regulation of MCL-1.

Objective　To identify aldosterone synthase gene-CYP11B2 mRNA expression in normal and fibrotic liver in rats and evaluate the curative effect of antisterone. Methods　160 Wistar rats weighing about 250?g were divided into 4 groups. In the model group (n=40), the rats were injected with 40% CCl4 (0.25?ml/100?g) subcutaneously three times a week. In the antisterone group (n=40), the rats were injected with 40% CCl4 (0.25?ml/100?g) subcutaneously three times a week. Antisterone equivalent to 20?mg*kg-1*d-1 was given intragastrically (ig). In the malotilate group (n=40), the rats were injected with 40% CCl4 (0.25?ml/100?g) subcutaneously three times a week. Malotilate equivalent to 50?mg*kg-1*d-1 was given ig. In the control group (n=40), the rats were injected with olive oil only. After 2,4,6,8 and 10 weeks, animals were sacrificed, and morphological examination was carried out. The area of collagen was examined with an Image Analyse System. Expression of the aldosterone synthase gene, CYP11B2 mRNA, in fibrotic and normal liver was detected by means of reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization. Results　In situ hybridization and RT-PCR showed that the expression of CYP11B2 mRNA, which localized in the endoplasm of hepatic stellate cells (HSCs), was up regulated when fibrogenesis occurred. Histological observation indicated that the grade of fibrosis and the area of collagen in the antisterone group were less than those in model group before 6 weeks (P<0.05). There was no significant difference between the antisterone and malotilate groups (P>0.05). After that, however, the grade of fibrosis and the area of collagen in the antisterone group were higher than those in the malotilate group (P<0.05). There was no significant difference between the antisterone and model groups (P>0.05). Conclusions　The expression of CYP11B2 mRNA is up regulated in fibrotic liver. Antisterone can have a partial fibrogenesis-inhibiting effect in the early stages.

OBJECTIVETo evaluate the progression in morphologic changes of lungs in SARS patients.

METHODSFour cases of SARS with lung tissue samples available (including one for ultrastructural examination) were enrolled into the study. Histochemical study for VG, Masson, reticulin, orcein, PAS, sirius red stains and immunohistochemical study for vimentin, desmin, smooth muscle actin, HHF-35, CD34, F8, collagen types I and III were also performed.

RESULTSAccording to the morphologic changes, lung lesions in SARS were subcategorized into 3 phases: acute exudative inflammation, fibrous proliferation and the final fibrotic stage. Two cases belonged to the acute exudative phase, in which the course was less than 20 days. The principal lesions consisted of acute alveolar exudative inflammation, hyperplasia of alveolar epithelium, necrosis, alveolar hyaline membrane formation, alveolar desquamation and focal fibroplasia. The acute exudative protein was PAS-positive. There was an increase in reticulin fiber formation. The reactive fibroblasts were highlighted by desmin and vimentin. One case belonged to the fibroproliferative stage, in which the course was around 25 days. Major lesions included proliferative interstitial pneumonia with early pulmonary fibrosis. There was also evidence of organizing pneumonia, with an increase in reticulin fiber formation, which had a glomeruloid appearance on special stain. The mesenchymal cells showed either myofibroblastic (which expressed desmin, HHF-35, smooth muscle actin and vimentin) or fibroblastic (which expressed vimentin only) differentiation. Fibroelastosis and fibroplasia was also noted. The remaining case belonged to the fibrotic stage, in which the course was around 75 days. The main features included diffuse fibrosis and honeycomb change, which were highlighted by sirius red stain. Immunohistochemistry showed mainly types I and IV collagen fibers. In all lesions, there was also an increase of number of CD68-positive macrophages.

CONCLUSIONSThe morphologic progression in lungs of SARS patients is characterized by the development of increased fibrosis. The primitive mesenchymal cells, hyperplastic alveolar epithelial cells and macrophages play an important role in the pathogenesis.

Actins ; metabolism ; Adult ; Collagen Type I ; metabolism ; Desmin ; metabolism ; Humans ; Lung ; metabolism ; pathology ; Male ; Middle Aged ; Pulmonary Fibrosis ; pathology ; Severe Acute Respiratory Syndrome ; metabolism ; pathology ; Vimentin ; metabolism

Objective To determine the distribution of HCV genotypes in patients with chronic hepatitis C,study the distribution of genotype in different gender and the relationship between genotypes and serum HCV-RNA levels.Methods Two hundred and six cases of HCV RNA positive patients(all with relevant clinical data) receiving pegylated interferon therapy were collected from May to December 2010.HCV RNA was detected in 206 hepatitis C patients from 40 hospitals in China by Roche Cobas AmpliPrep/Cobas TaqMan HBV test,and genotype was determined by Abbott RealTime HCV G enotype Ⅱ .The distribution of genotypes in the gender was analyzed by chi-square test analysis.The relationship between genotypes and serum HCV RNA levels was detected by single factor analysis and two independent sample t test analysis.Results There were seven different subtypes of HCV in 206 samples,including genotype 1,7 cases(3.4% ,7/206); genotype 1a,2 cases(1.0%,2/206); genotype 1b,123 cases (59.7 %,123/206); genotype 2,32 cases(15.5 %,32/206); genotype 3,27 cases(13.1%,27/206); genotype 6,6 cases(2.9% ,6/206) ;genotype 1/6,5 cases(2.4% ,5/206) ;genotype 2/4,1 cases(0.5%,1/206).There was no significant difference between HCV genotype and gender in 132 cases with genotype 1 and 65 cases with non-genotype 1(genotype 2,3,6) (x2 = 0.000,P ＞ 0.05).There was significant association between quantity of HCV RNA and genotype in 188 patients with HCV(F = 3.371,P＜ 0.01).The 197 patients with HCV single genotype were divided into five groups in terms of region(East,South,West,North and Center).There was no significant difference between HCV genotype 1 and non-genotype 1 in the five groups(x2 = 5.840,P ＞ 0.05).Conclusions It is suggested that HCV 1 b is the most prevalent type in China,followed by HCV 2.There is no significant difference between HCV genotype and gender.The levels of HCV RNA with genotype 1b are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 2 are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 6 are significantly higher than those with genotype 3.

Objective To express,purify and identify recombinant hepatitis E virus(HEV) pb166-GST fusion protein using GST gene fusion system and investigate its potential role in researching Hepatitis E diagnostic antigen field.Methods The recombinant E.coli BL21 performed by our own laboratory was used to induce the HEV pb166-GST expression with IPTG.The products were purified by BD Biosience GST purifying system.The specific expression was identified by SDS-PAGE and Western blot.The experiment conditions and results were described and analysed.Results The resolved HEV pb166-GST fusion protein on SDS-PAGE showed a major band at position of 43 kD.The expressed proteins had a single expected band after purify and the protein was recognized by anti-GST antibody on PVDF membrane.Conclusion The recombinant HEV pb166-GST fusion protein is expressed in recombinant E.coli BL21 efficiently in this way,and might be used as a candidate for diagnostic antigen of HEV.

Objective To investigate the curative effect of acute severe brain injury complicated ARDS, Methods 31 patients who had acute severe brain injury complicated ARDS were divided into two groups:A group was early discovery of ARDS and given treatment.B group was late discovery of ARDS and treated late.Then the curative effects were compared.Results A group was significantly higher than B group in blood gas analysis(P

Objective To explore the clinical pathway (CP) in gynecology laparoscopic surgery patients in the implementation effect and feasibility. Methods 200 patients with gynaeeological laparoscope operation were analyzed retrospectively from June to July 7 in 2008. The CP, and the relevant guidelines were applied to clinics after clinical experts' confirmation. 96 cases before CP (control group) and 100 cases after CP (experimental group ) were collected and analyzed including hospitalized days and expenses, patient satisfaction, and variation factors of implementing CP. Results The average of hospitalized days were (6.47± 2.53 ) d in experimental group and 8.38 ± 2.01 d in the control group. The hospitalization expenses were $ (9 080.62 ± 139.12 ) in experimental group and $ ( 9 655.08 ± 186.31 ) in the control group ( P < 0. 05 ). Variation factors of CP were mainly included medical personnel and patients with subjective understanding on the disease. Conclusions CP is helpful to reduce the hospitalized days, shorten the medical expenses, and ensure the quality of medical care, improve work efficiency, even were welcomed by the nurse.