Effect of prenatal exposure to passive smoking on leptin of neonatal cord blood and birth weight in a rural area of Yunnan Province / 环境与职业医学

Background Previous studies have shown that active smoking during pregnancy can reduce the level of neonatal cord blood leptin, and thereby affect birth weight. However, few studies have studied the association of passive smoking during pregnancy with leptin in neonatal cord blood and birth weight. Objective To explore the effects of passive smoking in varied pregnancy stages and entire pregnancy on neonatal cord blood leptin level and birth weight in a certain rural area of Yunnan, and potential mediating role of cord blood leptin. Methods Based on a prospective prenatal cohort study conducted in Xuanwei County, Yunnan Province, a total of 545 mother-infant pairs were included in this study from early pregnancy enrollment to delivery. The demographic information and reproductive history of the subjects were collected by questionnaire. The urine samples of pregnant women in the first, second, and third trimesters of pregnancy were collected during regular prenatal examinations. Umbilical cord blood samples were collected from newborns at birth. The concentration of urine cotinine (UC) was measured by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). According to the results of UC level during pregnancy, the study subjects were divided into three groups: negative group (<LOD), low exposure group (LOD-M), and high exposure group(>M). The level of leptin in cord blood was detected by ELISA. Multiple linear regression was used to analyze the effect of passive smoking on umbilical cord blood leptin in newborns during pregnancy. Path analysis was used to explore the relationship among passive smoking during pregnancy, neonatal cord blood leptin, and birth weight. Results The average exposure rate of passive smoking during pregnancy was 87.28%, and the exposure rate for entire pregnancy was 76.88%. The median concentration of leptin in neonatal cord blood was 4.17 μg·L−1. After adjusting for maternal age, ethnicity, educational level, pre-pregnancy body mass index (BMI), gestational weight gain, parity, annual household income, infant sex, and birth weight, we found that low level (b=−3.388, P=0.001) and high level (b=−2.738, P=0.006) of passive smoking in the first trimester of pregnancy had negative associations with leptin concentration of cord blood by multiple linear model. The path analysis results showed that passive smoking in the first trimester and pre-pregnancy BMI directly affected leptin levels, and the sizes of direct effects were −0.073 and −0.087 (both P<0.05) respectively. Passive smoking in late pregnancy, gestational weight gain, premature, newborn girls, parity, and pre-pregnancy BMI directly affected birth weight, and the sizes of direct effects were −0.063, 0.191, −0.301, −0.128, −0.121, and 0.167 (all P<0.05), respectively. No mediating role of leptin was found in the effect of passive smoking on neonatal birth weight. Conclusion Passive smoking exposure during pregnancy is common among rural women in Yunnan Province. Passive smoking in the first trimester may be key in decreasing the leptin level of neonatal cord blood. Passive smoking in third trimester may lead to a decrease in birth weight. No evidence shows that leptin mediates the relationship between passive smoking and birth weight.

Role of Wnt/β-catenin Signaling Pathway in Seborrheic Alopecia and Its Intervention with Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Seborrheic alopecia is a chronic dermatological disease caused by multiple factors. It occurs frequently in young and middle-aged men aged 20-30 years. The main clinical manifestations are greasy hair， itching， excessive dandruff， receding hairline， sparse hair on the top of the head， and progressive hair loss in the frontotemporal area. Seborrheic alopecia is not fatal， but it affects the appearance of patients， seriously harming their self-esteem and bringing great psychological distress to them. The Wnt/β-catenin signaling pathway widely exists in multicellular eukaryotes and is a basic growth regulatory pathway which regulates cell proliferation and differentiation， maintains stem cells activity and organ homeostasis， and affects cell migration. At present， it has been reported in China and abroad that Wnt/β-catenin signaling pathway is closely related to the occurrence and development of seborrheic alopecia and the action mechanism of drugs. Traditional Chinese medicine （TCM） has multi-component， multi-target， and multi-pathway advantages， and it can promote the formation of hair follicle laminae， the proliferation and differentiation of hair follicle stem cells， and the periodic changes in hair follicles by regulating the Wnt/β-catenin signaling pathway， thereby alleviating seborrheic alopecia. This article reviewed the relationship of Wnt/β-catenin signaling pathway and its key target protein factors with seborrheic alopecia to clarify the important role of Wnt/β-catenin signaling pathway in seborrheic alopecia. At the same time， the TCM that targeted the Wnt/β-catenin signaling pathway to relieve seborrheic alopecia were summarized， so as to provide reference for the treatment of seborrheic alopecia and further development of new drugs.

Prenatal pyrethroid pesticides exposure and neurodevelopment of 2-year-old children: a birth cohort study / 中华预防医学杂志

Objective: To investigate the influence and critical windows of prenatal exposure to pyrethroid pesticides (PYRs) on neurodevelopment of 2-year-old children. Methods: The subjects of this study were derived from the Xuanwei Birth Cohort. A total of 482 pregnant women who participated in the rural district of Xuanwei birth cohort from January 2016 to December 2018 were included. Maternal urinary concentrations of PYRs metabolites during 8-12 gestational weeks, 20-23 gestational weeks and 32-35 gestational weeks were measured with ultra high performance liquid chromatography system coupled with a tandem mass spectrometry detector. Child neurodevelopment was evaluated with the Bayley Scales of Infant and Toddler Development-Third Edition at 2 years of age. Multivariate linear regression models and binary logistic regression models were used to assess the association between PYRs exposure during pregnancy and children's neurodevelopment. Results: A total of 360 mother-child pairs had complete data on maternal urinary PYRs metabolites detection and children's neurodevelopment assessment. The detection rate of any one PYRs metabolites during the first, second and third trimester were 93.6% (337/360), 90.8% (327/360) and 94.2% (339/360), respectively. The neurodevelopmental scores of Cognitive, Language, Motor, Social-Emotional, and Adaptive Behavior of 2-year-old children were (102.3±18.9), (100.2±16.3), (102.0±20.3), (107.8±23.3) and (85.8±18.6) points, respectively. After controlling for confounding factors, 4-fluoro-3-phenoxybenzoic acid (4F3PBA, one of PYRs metabolites) exposure in the first trimester reduced Motor (β=-5.02, 95%CI: -9.08, -0.97) and Adaptive Behavior (β=-4.12, 95%CI:-7.92, -0.32) scores of 2-year-old children, and increased risk of developmental delay of adaptive behavior (OR=2.07, 95%CI:1.13-3.82). Conclusion: PYRs exposure during the first trimester of pregnancy may affect neurodevelopment of 2-year-old children, and the first trimester may be the critical window.

Analysis of Effect of Realgar with Different Compatibility Proportions on Transitional Constituents of Indigo Naturalis in Rat Serum Based on UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To investigate the effect of different dose of Realgar compatible with Indigo Naturalis on the transitional constituents of Indigo Naturalis in rat serum based on the compatibility of Qinghuangsan.Method:Indigo Naturalis test solution, the drug-containing serum of three different proportions of Qinghuangsan (10 g of Indigo Naturalis compatible with 52.5, 105, 210 mg of Realgar for group A, B and C, respectively) and blank serum were detected by UPLC-Q-TOF-MS/MS, in combination with the chemical components identified in Indigo Naturalis test solution, the differences of transitional constituents of Indigo Naturalis in rat serum from the group A, B and C were analyzed. HL-60 cells (human leukemia cells) were treated with the three groups of Qinghuangsan drug-containing serum and the effect of drug-containing serum on the activity of HL-60 cells was detected by cell counting kit-8 (CCK-8) assay.Result:A total of 19, 22, 25 of transitional constituents were detected in Qinghuangsan drug-containing serum from group A, B and C, respectively. The three groups of drug-containing serum all contained 5 prototype components from Indigo Naturalis test solution, including tryptanthrin, indigo, indirubin, 2-aminobenzoic acid and N-phenyl-2-naphthylamine, respectively. The results of CCK-8 assay showed that Qinghuangsan drug-containing serum of group C had the strongest inhibitory effect on HL-60 cells.Conclusion:After fixed Indigo Naturalis dose, with the increase of Realgar dose, the transitional constituents in rat serum increase and the inhibitory effect on HL-60 cells also gradually enhances, which indicates that Realgar may promote the absorption of active components in Indigo Naturali in vivo, thus enhance the efficacy, further explains the compatibility law and pharmacodynamic material basis of different proportions of Realgar and Indigo Naturalis.

Determination of voriconazole in dog plasma by lc-ms/ms and its toxicokinetics / 中国药学杂志

OBJECTIVE: To establish a method for the determination of voriconazole in dog plasma and investigate its toxicokinetics. METHODS: After protein precipitation with acetonitrile, voriconazole and fluconazole were separated on an Agilent Poroshell120 EC-C18 column (2.1 mm×50 mm, 2.7 μm), with acetonitrile and water (0.1% FA) as the mobile phase at a flow rate of 0.3 mL·min-1. Detection was carried out by the electrospray positive ionization mass spectrometry in the multiple reacion monitoring (MRM) mode. The MRM transitions of m/z 335.1→281.3 and m/z 307.1→220.0 were used to quantify voriconazole and fluconazole, respectively. Thirty Beagle dogs received intravenous infusion of voriconazole at low, medium and high doses (1, 3, 6 mg·kg-1·d-1) once a day for 12 weeks. RESULTS: The calibration curve was linear over 10-10 000 ng·mL-1 . RSD was less than 15%, and the accuracy was within the range of 85%-115%.The exposure of voriconazole to females was significantly higher than that of males in Beagle dogs. When the dose was 1-6 mg·kg-1·d-1, the exposure of voriconazole to beagle dogs increased with the increase of dose, and the drug did not accumulate. CONCLUSION: The method can be applied to the determination of voriconazole in dog plasma, and is suitable to the toxicokinetics study of voriconazole.

Determination of (S)-Pantoprazole Sodium in Dog Plasma by LC-MS/MS and Its Toxicokinetic Study / 中国药学杂志

OBJECTIVE: To establish an LC-MS/MS method to determine (S)-pantoprazole sodium in dog plasma and investigate its toxicokinetics. METHODS: After protein precipitation with acetonitrile, the analyte and internal standard were separated on CHIRALCEL OJ-RH column (4.6 mm ×150 mm, 5 μm) with acetonitrile-water (28∶72) as mobile phase eluted at a flow rate of 0.6 mL·min-1. Detection was carried out by electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode. The MRM transitions of m/z 384.0/199.8 and m/z 180.0/110.0 were used to quantify (S)-pantoprazole sodium and phenacetin, respectively. Beagle dogs were intravenously given (S)-pantoprazole sodium for 4 weeks at low, medium, and high dosages (10, 20, 40 mg·kg-1·d-1). RESULTS: The calibration curve was linear over the concentration range of 50-30 000 ng·mL-1. The RSDs were less than 15%, and the accuracy was in the range of 85%-115%. The AUC0-4 h and ρmax of (S)-pantoprazole sodium were proportional to the dosages. CONCLUSION: The established method can be applied to the determination of (S)-pantoprazole sodium in plasma of dogs and is suitable for the toxicokinetic study.

Preparation and antimicrobial effect of aromatic, natural and bacteriostatic foot wash with skin care / 中国中药杂志

To prepare the aromatic, natural and bacteriostatic foot wash with skin care and research the inhibition effect on the different bacteria and pathogenic fungus which cause dermatophytosis. It was prepared by using Sophoraflavescens and Dictamnus dasycarpus as materials with the addition of Aloe extract, essential oil, surfactant, etc. The antifungal and antibacterial activity was researched by the levitation liquid quantitative method. The foot wash smelled faintly scent. The use of this product can produce a rich foam. The inhibitory rate were all more than 90%. The preparation process of the foot wash was simple. It has obviously bacteriostatic and fungistatic effect.

The expressions of plasma hsa-miR-150 and hsa-miR-375 in patients with colorectal cancer and its clinical significance / 肿瘤

Objective: To select the plasma miRNA (microRNA) associated with the clinical features of patients with colorectal cancer. Methods: Differential expression of plasma miRNA was detected by TaqMan® Human MicroRNA Array from 7 patients with colorectal cancer and 6 healthy volunteers. The results of TaqMan® Human MicroRNA Array were validated by using plasma samples from 29 patients with colorectal cancer and 20 healthy volunteers by real-time fluorescence quantitative-PCR, and the relationship of differential expression with clinical features was analyzed. Results: In the condition of fold change > 1.0 and P < 0.05, sixteen plasma-specific miRNAs were identified from patients with colorectal cancer. Hsa-miR-125b, hsa-miR-375, hsa-miR-150 and hsa-miR-206 validated by real-time fluorescence quantitative-PCR were consistent with those detected by TaqMan® Human MicroRNA Array, and the differential expression was statistically significant between colorectal cancer patients and healthy volunteers (P < 0.05). Furthermore, the expression of plasma hsa-miR-150 and hsa-miR-375 had a significant impact on tumor invasion and vascular cancer embolus. Conclusion: Plasma hsa-miR-150 and hsa-miR-375 are expected to be non-invasive biomarkers to give assistance in tumor staging and to evaluate the prognosis of colorectal cancer. Copyright © 2012 by TUMOR.

Identification of metabolites of nobiletin in rats using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry / 药学学报

In this study, metabolism of nobiletin in rats was studied using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). As a result, seven major metabolites were found in bile, urine and serum of rats. Three phase I products were assigned to be demethyl and di-demethyl products, and other four phase II products were assigned to be glucuronic and sulfonic conjugates. The four phase II metabolites were reported for the first time. Among the metabolites found in the present study, the glucuronic conjugates of demethyl-nobiletin played a predominant role in the metabolic pathway, indicating that its potential role for glucuronidation-related factors, such as gene polymorphism, drug-drug interaction, etc., in changing the active and toxic effect of nobiletin and that it should be paid more attention in further development.

A comparison of NK cell subsets between rhG-CSF mobilized peripheral blood grafts and bone marrow grafts from healthy donors / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the difference in NK cell subsets between recombination human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood grafts (G-PB) and bone marrow grafts (G-BM) from healthy donors.</p><p><b>METHODS</b>From July 2009 to September 2009, G-PB and G-BM from 28 related donors were collected to analyze lymphocytes, NK cells and NK cell secretion of interferon-γ (IFN-γ, NK1), interleukin-13 (IL-13, NK2), transforming growth factor-β (TGF-β) and interleukin-10 (IL-10, NKr) by flow cytometry.</p><p><b>RESULTS</b>The percentage of lymphocytes in G-PB was significantly higher than that in G-BM (P < 0.01). The proportions of NK cells and NK1, NK2, NKr subsets among lymphocytes were significantly higher in G-BM than in G-PB (P < 0.05), and so were the percentage of NK2, NKr cells among NK cells (P < 0.01), but no significant difference in the percentage of NK1 cells among NK cells between G-PB and G-BM. The ratios of IL-13 and IFN-γ, of TGF-β and IFN-γ, or of IL-10 and IFN-γ in G-BM were significantly higher than those in G-PB (P = 0.010, 0.002, or 0.000, respectively).</p><p><b>CONCLUSION</b>The increased proportion of NK2 and NKr in G-BM might be helpful to explain the lower immunoreactivity of G-BM than that of G-PB, although the proportion of NK1 in G-BM and G-PB is similar.</p>

Accuracy of a continuous glucose monitoring system in detection of blood glucose during oral glucose tolerance test / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the accuracy of continuous glucose monitoring system (CGMS) during oral glucose tolerance test (OGTT) in the detection of blood glucose changes in glucose stress condition.</p><p><b>METHODS</b>Forty-nine out-patients with fasting plasma glucose of 3.9-11.0 mmol/L underwent continuous blood glucose monitoring using CGMS for 3 days, and OGTT was conducted on the third day. The venous blood glucose was measured at 0, 30, 60, 90, and 120 min after oral glucose intake, and the accuracy of CGMS during OGTT was evaluated.</p><p><b>RESULTS</b>The correlation indices between CGMS values and the venous blood glucose values during the entire OGTT and in phases of stable, rapidly rising and falling glucose levels were 0.928, 0.901, 0.924 and 0.902, respectively (P<0.001). Clarke error-grid analysis showed that more than 95% of the measured results fell into the A and B zones.</p><p><b>CONCLUSION</b>CGMS values show good consistency with venous blood glucose values measured during OGTT. CGMS is accurate in detection of rapidly changing blood glucose during OGTT.</p>

Effects of rhG-CSF mobilization on Th17 cells in donors' peripheral blood and bone marrow grafts / 中国实验血液学杂志

The study was aimed to explore the effects of recombination human granulocyte colony-stimulating factor (rhG-CSF) on Th17 cells in donors' peripheral blood (GPB) and bone marrow grafts (GBM). 25 healthy donors were injected subcutaneously with rhG-CSF 5 µg/(kg·d) for 5 consecutive days. GBM and GPB were harvested after injection on day 4 and 5 respectively. Some of these donors' steady-state bone marrow (SSBM) and steady-state peripheral blood (SSPB) were harvested before rhG-CSF injection. The changes of IL-17 secreted by T cells in donor BM and PB before and after mobilization were detected by flow cytometry. The results showed that the ability to secrete IL-17 from CD4(+) T cells and CD8(+) T cells in GBM was significantly lower than those in SSBM (GBM vs SSBM Th17/CD4(+) T, 0.74% ± 0.27% vs 1.78% ± 1.19%, p < 0.05; Tc17/CD8(+)T, 0.19% ± 0.16% vs 0.36% ± 0.37%, p < 0.05), changes in peripheral blood and bone marrow were same (GPB vs SSPB Th17/CD4(+) T, 1.82% ± 0.91% vs 3.26% ± 1.89%, p < 0.01; Tc17/CD8(+) T, 0.21% ± 0.17% vs 0.44% ± 0.28%, p < 0.01). The ratios of Th17/CD4(+) T and Tc17/CD8(+) T in GPB were higher than in GBM (p < 0.05, p < 0.01). It is concluded that rhG-CSF in vivo can inhibit the generation of Th17 cells both in bone marrow and peripheral blood grafts, and it may be partial reason for GPB/GBM mixed transplantation without increasing the GVHD incidence.

The association between common genetic variation in the FTO gene and metabolic syndrome in Han Chinese / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Genome-wide association studies for type 2 diabetes mellitus (T2DM) identified FTO gene as a locus conferring increased risk for common obesity in many populations with European ancestry. However, the involvement of FTO gene in obesity or T2DM related metabolic traits has not been consistently established in Chinese populations. The objective of this study was to investigate the association of FTO genetic polymorphisms with metabolic syndrome (MetS) in Han Chinese.</p><p><b>METHODS</b>We tested 41 FTO single nucleotide polymorphisms (SNPs) for association between FTO and MetS-related traits. There were a total of 236 unrelated subjects (108 cases and 128 controls), grouped according to the International Diabetes Federation (IDF) criteria.</p><p><b>RESULTS</b>Of the 41 SNPs examined, only SNP rs8047395 exhibited statistical significance (P = 0.026) under a recessive model, after Bonferroni adjustment for multiple testing (OR 1.64, 95%CI 1.11-2.42; P = 0.014). The common distributions of this polymorphism among Chinese--with a minor allele frequency (MAF) of 36% in the control group versus 48% in the MetS group--greatly improved our test power in a relatively small sample size for an association study. Previously identified obesity- (or T2DM-) associated FTO SNPs were less common in Han Chinese and were not associated with MetS in this study. No significant associations were found between our FTO SNPs and any endophenotypes of MetS.</p><p><b>CONCLUSIONS</b>A more common risk-conferring variant of FTO for MetS was identified in Han Chinese. Our study substantiated that genetic variations in FTO locus are involved in the pathogenesis of MetS.</p>

Changes of insulin resistance and islet beta cells function in subjects with high-normal blood pressure / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the changes of insulin resistance and islet beta cells function in subjects with euglycemia and high-normal blood pressure.</p><p><b>METHODS</b>Total 423 subjects were divided into normal blood pressure group and high-normal blood pressure group. Body height, weight, waist and hip circumference, and biochemical data were measured. Homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI)-composite, and first-phase (1 PH) Stumvoll index were calculated. Results Waist circumference, total cholesterol, triglyceride, low-density lipoprotein cholesterol, HOMA-IR were significantly higher and IPH Stumvoll index and ISI-composite were significantly lower in high-normal blood pressure group than in normal blood pressure group (P < 0.05). Systolic blood pressure (SBP) was positively correlated with HOMA-IR (r = 0.122) and negatively correlated with 1PH Stumvoll index (r = -0. 159) and ISI-composite (r = -0.131) (P < 0.05). SBP and triglyceride were independent factors for IPH Stumvoll index.</p><p><b>CONCLUSION</b>Insulin resistance and islet dysfunction may exist in subjects with high-normal blood pressure.</p>

Association of ghrelin polymorphisms with metabolic syndrome in Han Nationality Chinese / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the association of ghrelin gene polymorphisms with metabolic syndrome in Han Nationality Chinese.</p><p><b>METHODS</b>A total of 240 patients with metabolic syndrome and 427 adults aged above forty years were recruited. Genotypes were determined by polymerase chain reaction and restriction fragment length polymorphism analysis.</p><p><b>RESULTS</b>The allelic frequency of the Leu72Met polymorphism was 17.3% in the patient group and 11.9% in the control group (chi2 = 7.36, P = 0.007). Metabolic syndrome was more prevalent among carriers of the Met72 variant (43.8 vs 33.1%, age- and sex-adjusted odds ratio = 1.57, P = 0.01). No Arg51Gln variants were found in our study subjects.</p><p><b>CONCLUSION</b>Rather than being associated with its individual components, Leu72Met polymorphism is associated with metabolic syndrome in the Han Nationality Chinese. Arg51Gln polymorphism is rare in the Han Nationality Chinese.</p>

Identification of a regulatory single nucleotide polymorphism in the adiponectin (APM1) gene associated with type 2 diabetes in Han nationality / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To identify the genetic defects of the the adiponectin (APM1) gene that contribute to the development of type 2 diabetes (T2DM) and determine the functional single nucleotide polymorphisms (SNPs) in the APM1 gene associated with T2DM in Han nationality.</p><p><b>METHODS</b>The APM1 gene 5'-UTR was screened by direct sequencing to identify common polymorphisms. Identified SNPs were genotyped in 585 nondiabetic controls, 278 subjects with impaired glucose intolerance (IGT) and 212 patients with T2DM. The functions of SNPs in the regulatory region were assessed by reporter gene assay. Possible association between SNPs and plasma APMI levels or metabolic parameters was statistically assessed.</p><p><b>RESULTS</b>Three SNPs were identified in the APM1 gene 5'-UTR. A case-control study revealed that SNP -11377 G/C had significant differences in allele frequencies between T2DM patients and nondiabetic controls (G 0.314/C 0.686 vs. G 0.265/C 0.735, P=0.03). Haplotype analysis of three SNPs in the APM1 gene showed that no significant association of haplotypes with T2DM. IGT was detected in the present study. Reporter gene assay showed that SNP did not influence the transcription efficiency in the 3T3-L1 cell line.</p><p><b>CONCLUSION</b>SNP -11377 G/C in the proximal promoter region of the APM1 gene contributes to the development of T2DM in Han nationality but may not be a functional SNP in the APM1 gene.</p>

Studies on the chemical constituents in root of Coleus forskohlii / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in root of Coleusforskohlii.</p><p><b>METHOD</b>The chemical constituents were isolated by column chromatography. The structures were elucidated on the basis of IR, MS, 1H-NMR, 13C-NMR and 2D-NMR experiments.</p><p><b>RESULT</b>Six compounds were obtained and the structures were identified as 14-deoxycoleon U (1), demethylcryptojaponol (2), alpha-amyrin (3), betulic acid (4), alpha-cedrol (5) and beta-sitosterol (6).</p><p><b>CONCLUSION</b>Compounds 1 and 2 were isolated from the genus Coleus for the first time. Compounds 3 and 4 were isolated from C. forskohlii for the first time.</p>

Gene cloning and expression of serine protease SFP2 from Streptomyces fradiae var. k11 / 生物工程学报

Extracellular serine protease SFP2 from Streptomyces fradiae var. k11 with high feather-degrading activity was purified. The partial amino acid sequences of internal peptide of purified SFP2 were determined, and the partial gene encoding SFP2 was cloned by PCR using the degenerate primers designed according to the amino acid sequences. Complete sfp2 gene was cloned by screening the genomic DNA library of Streptomyces fradiae var. k11. The Open Reading Frame of sfp2 including pre- pro-enzyme is 924bp long (EMBL Accession number: AJ784940). The signal peptide sequence is as long as 114bp, the precursor sequence is 810bp and the mature enzyme is 576bp long, encoding 191 amino acid resides with the putative molecular weight of 19.112kD. In E. coli and Bacillus subtilis, the two sequences encoding SFP2 pro-enzyme and mature enzyme were both expressed successfully. The pro-enzyme expressed had normal biological function and its mature product had normal enzymatic activity.