Effects of glucose concentration and pH\|control upon production of exopolysaccharides of Grifola frondosa was studied.The results showed that glucose at concentration of 5% was favorable to exopolysaccharides production,and the pellets benefited from the controlled start pH as well as the controlled final pH of 3 5～4 0 to produce exopolysaccharides.Attempt on the exopolysaccharides production with recycled pellets in the saccharoid solution was also made in this work.

Objective To detect the change and clinical significance of homocysteine (Hcy),hypersensitivity C response protein (hs-CRP) and hypersensitivity troponin I (hs-TnI) in patients with acute myocardial infarction (AMI).Methods Ninety-six cases of AMI were selected as the research group and 60 cases of coronary heart disease without AMI were enrolled in the control group.Blood samples were collected to measure the contents of serum Hcy,hs-CRP and hs-TnI.ROC curve was drawn and Logistic regression model was established to analyze the role of each index alone and combined detection in the diagnosis of AMI.Results The levels of Hcy,hs-CRP and hs-TnI in the research group were significantly higher than those in the control group (Hcy: (29.29±7.65) mol/L vs.(17.23±4.68) mol/L;hs-CRP: (15.47±5.01) mg/L vs.(9.21±3.15) mg/L;hs-TnI: (40.88±9.18) ng/mL vs.(7.34±2.12) ng/mL,t=2.78,8.66,34.36,P<0.05).The area under the ROC curve of Hcy,hs-CRP,hs-TnI were 0.802 (95%CI(0.729~0.874)),0.71(95%CI(0.62~0.792)),0.929 (95%CI(0.891~0.967)),respectively.The area under the combine detection curve was 0.971 (95%CI,0.950~0.992).The sensitivity of Hcy,hs-CRP and hs-TnI in the diagnosis of AMI were 79%,57%,87%,respectively,the specificity of the three groups were 72%,70%,90%,the sensitivity and specificity of combined detection were 96% and 88%.Conclusion Combined detection of Hcy,hs-CRP and hs-TnI plays a better role in the diagnosis of AMI than that of Hcy,hs-CRP,hs-TnI alone and has a high specificity and sensitivity in the diagnosis of AMI.

AIM: To study the purification of Naomaitong Granules(Radix et Rhizoma rhei,Rhizoma chuanxiong,Radix Puerariae lobatae.etc) by macroporous absorption resin. METHODS: Naomaitong Granule was purificated by macroporous absorption resin AB-8.UV spectrothotometry was used to determine the contents of total anthraquinones,total ginsenosides,total alkaloids,and the content of puerarin was determined by HPLC.The technic of purification was optimized according to the content above. RESULTS: The optimized technological conditions consisted of eoncentranon of original sample 120 mg/mL,the diameter and height was in proportion of 1∶10,the ratio of maximum adsorption to resin volume was 1∶6,water elution was 2B multiple of resin volume,8B multiple of resin volume 50% alcohol was the elution. CONCLUSION: AB-8 macroporous absorption resin can be used to purify Naomaitong Granules.

Objective To identify the genetic mechanism of fetuses with short limbs deformity.Methods From Aug.2008 to Aug.2011,ten fetuses with obvious short limbs were found in ultrasound screening performed at 18-24 and (or) 30-32 gestational weeks and underwent artificial induced labor with the patient' consent.Amniotic fluid or cord blood of the fetuses was collected for karyotyping analysis and detection of mutation point of fibroblast growth factor receptor 3 (FGFR3)gene by polymerase chain reaction and gene sequencing.One fetus (case 3) who presented with achondrogenesis underwent sequencing of SLC26A2 and Trip11 gene meanwhile.Results Among the 10 fetuses with short limbs deformity,five cases were found during second trimester and five during third trimester.Nine cases were identified as normal karyotype and one was chimera (46,XY/45,XY,- 18).One fetus carried a rare FGFR3 mutation of c.1108G＞T (G370C) and was diagnosed as thanatophoric dysplasia at 21+3 weeks.Three fetus carried c.1138G＞A (G380R) mutation and were diagnosed as achondroplasia.These four families had low recurrent risk because no gene mutations were found in the parents.Three mothers of these four fetuses were pregnant again and had normal neonates now.No mutations were found in all gene sequencing in case 3.Conclusions Karyotyping analysis and sequencing of FGFR3 gene could find causative gene mutations and provide genetic counselling and prenatal diagnosis for some fetuses with short limbs deformity.In the third trimester,achondroplasia is the most possible diagnosis when short limbs fetus is found by ultrasound.

Objective To investigate the value of detection of fetal cell-free fetal DNA(cff-DNA)in maternal plasma in the prenatal diagnosis of chromosomal abnormalities.Methods The plasma from 3200 gravidas(singleton with 20.3 ± 3.8 gestational weeks)was collected from April 1st 2011 to May 30th 2012.They were divided into 3 groups:(1)To tally 1720 cases were included in the high-risk serological screening group,in which women were younger than 35 years and got high-risk results in serological screening;(2)To tally 1310 cases were included in the advanced age group,in which women's age was more than 35 years;(3)To tally 170 cases were included in the supplementary group,in which women were younger than 35 years and got low-risk results in serological screening,or women who didn't take serological screening tests.All the 3030 gravidas in group 1 and 2 didn't take invasive prenatal diagnosis because of fear of abortion or short of prenatal diagnosis.Cff-DNA were detected by next generation sequencing in Shenzhen BGI Genomics Center for clinical laboratory.Amniocentesis and karyotype analysis were provided to the positive cases and women with negative results were followed-up by telephone.Results(1)The 3200 cases took cff-DNA detection,and 31 cases got positive results,including 27 cases of trisomy 21 and 4 cases of trisomy 18.Sixteen cases of trisomy 21 and 1 case of trisomy 18 were in the high-risk serological screening group.7 cases of trisomy 21 and 2 cases of trisomy 18 were in the advanced age group.Four cases of trisomy 21 and 1 case of trisomy 18 were in the supplementary group.(2)And the 84％(26/31)cff-DNA detecting positive cases received amniocentesis.In the 27 trisomy 21 positive cases,23 received amnioeentesis and got karyotype of 47XN,+ 21,with the diagnostic accordance rate of 100％.In the 4 cases who didn't take karyotype analysis,fetal anomaly(ventricular septal defect,dextrocardia and choroid plexus cyst)was found in 1 case before 20 gestational weeks;intrauterine fetal demise happened in 1 case before getting the result;2 other cases who already had healthy children took abortion in the local hospital without taking amniocentesis.In the 4 trisomy 18 positive cases,3 took amniocentesis,2 of which were trisomy 18 and took abortion,the other was chimera(46,XN/47,XN,+ 18)with only 2％ cells of trisomy 18,with no malformation found after delivery.Hypoevolutism(3 weeks less than gestational week),general hydropsy and intrauterine fetal demise happened before the other case took amniocentesis.(3)Follow up of cff-DNA negative cases:until May 30th 2012,no Down's baby was found in the 1230 cases with cff-DNA test negative results.Conclusions(1)The non-invasive fetal trisomy test(NIFTY)by next generation sequencing is a safe,accurate and high throughput method for the prenatal diagnosis of trisomy-21.(2)Use NIFTY as a further screening for pregnant women with high-risk serological screening results could lower invasive prenatal diagnosis rate.(3)Cases with positive NIFTY test results should receive amniocentesis and karyotype analysis to confirm the diagnosis before abortion.

OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.

AIM: To establish an HPLC method for determing content of puerarin isoflavones in effective parts of Naomaitong Granules.METHODS: Gradient elution with methanol-water-glacial acetic acid was used as the mobile phase.The flow rate was 1.0 mL/min,and the detection wavelength was set at 250 nm.RESULTS: The average recoveries of puerarin,daizein-8-capiosy(1→6) glucoside,3-methoxyl-puerarin,daidzin,daidzein were 100.89%(RSD = 2.1%),100.23%(RSD = 1.06%),101.04%(RSD = 1.92%),99.82%(RSD = 2.02%),102.06%(RSD = 1.34%) respectively.CONCLUSION: The method is simple,accurate and can be used for quality control of the effective parts in Naomaitong Granules.

Objective To analyze the mutations of causal genes in sixteen Chinese patients with suspicious Bartter syndrome,and follow up their treatment results.Methods Mutations were identified by the next generation sequencing and the multiplex ligation-dependent probe amplification (MLPA).Clinical and biochemical features at the first presentation as well as follow-up results were reviewed.Results 15 different CLCNKB gene mutations were identified in sixteen patients with BS,including 11 novel ones.A novel missense mutation and a novel small deletion were found from SLC12A1 gene.A novel gross deletion was found in CLCNKA gene.A recurrent missense mutation was identified from BSND gene.The whole gene deletion mutation of CLCNKB gene was the most frequent mutation (32％),and the rate of gross deletion was up to 50 percent in this group of Chinese patients.The most common clinical manifestations were development retardation (15/16),polydipsia and polyuria (15/16).All of the patients were detected with hypokalemia,hypochloremia and metabolic alkalosis.Indomethacin treatment had significant improvement to the stature and weight restoration.Conclusion The present study has found 19 mutations,including 14 novel ones,which enriches the human gene mutation database (HGMD) and provides valuable references to the genetic counseling and diagnosis of Chinese population.

Aim To observe the antifibrogenic effect of resveratrol on mice with schistosomiasis japonica and its effect on Th1 and Th2 responses .Methods Forty-five mice infected with S.japonicum cercariae for 3 weeks were randomly divided into three groups named as infection group ( A) , resveratrol group ( B) and praz-iquantel group ( C) .Fifteen normal mice were taken as normal control group ( D) .In the 13th week post-infec-tion, all mice were sacrificed and the liver tissues were removed.Histopathological changes were observed in the liver of all groups .Splenocytes were prepared from spleens of mice with S.japonicum infection and the proportions of Th1 and Th2 cells in T cells were deter-mined by FACS respectively .RT-PCR was used to de-tect the relative IFN-γ,IL-13,TGF-βmRNA levels in liver tissue .Results After treatment , the degrees of liver fibrosis in groups B and C decreased in the 13th week post-infection ( P <0.01 ) .Compared to group A, the proportions of Th1 cells in group B significantly increased ( P<0.05 ) and the proportions of Th 2 cells in group B decreased significantly ( P <0.01 ) .The level of anti-SWA IgG 2 a in group B was significantly higher ( P<0.05) , while the anti-SEA IgG1 level in group B was lower ( P <0.01 ) than that in group A . The hepatic expression of IFN-γmRNA level in group B was higher than that in group A ( P<0.05 ) , and IL-13 ,TGF-βmRNA levels in group B were lower than in group A ( P<0.01 ) .Conclusion Resveratrol has an antifibrogenic effect through upregulating Th 1 cell re-sponse and downregulating Th 2 cell response in mice infected with Schistosoma japonicum.

Abstract: Our previous work revealed berberine can significantly enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans, which suggested that berberine has synergistic antifungal activity with fluconazole. Preliminary SAR of berberine needs to be studied for the possibility of investigating its target and SAR, improving its drug-likeness, and exploring new scaffold. In this work, 13-substitutited benzyl berberine derivatives and N-benzyl isoquinoline analogues were synthesized and characterized by 1H NMR and MS. Their synergetic activity with fluconazole against fluconazole-resistant Candida albicans was evaluated in vitro. The 13-substitutited benzyl berberine derivatives 1a-1e exhibited comparable activity to berberine, which suggested that the introduction of functional groups to C-13 can maintain its activity. The N-benzyl isoquinolines, which were designed as analogues of berberine with its D ring opened, exhibited lower activity than berberine. However, compound 2b, 2c, and 4b showed moderate activity, which indicated that berberine may be deconstructed to new scaffold with synergistic antifungal activity with fluconazole. The results of our research may be helpful to the SAR studies on its other biological activities.