Objective :To establish a RP-HPLC method for the determination geniposide in Yigan Liangcha.Methods Luna C18 (4.6 mm?250 mm,5 ?m)was applied,the mobile phases consisted of acetonitrile-water(15∶85),the flow rate was 1.0 mL?min-1 and the detection wavelength at 238 nm.Results The average recovery of geniposide was 98.1 %and RSD were 0.42 %(n=5).There is a good linear relationship within the range of 10.12～50.60 ?g of geniposide.Conclusion The method is convenient,sensitive,accurate and reproducible and can be used the quality control of the Yigan Liangcha.

Object To study the chemical constituents of the dried root of Fordia cauliflora Hemsl which is indigenous to Guangxi and commonly used in traditional medicine of Zhuang and Yao nationalities Methods The constituents were isolated with column chromatography and their structures were elucidated on the basis of physicochemical data and spectral analysis Results The three constituents were isolated and identified as 2 [3 methoxyphenyl] 4H furo[2,3 h] 1 benzoyran 4 one (Ⅲ), 3 methoxy 2 [3 hydroxyphenyl] 4H furo [2, 3 h] 1 benzopyran 4 one (Ⅳ) and pinnatin (Ⅴ) Conclusion The all of them are isolated from this plant for the first time Compound Ⅲ,Ⅳ are new and tentatively named as cauliflorin A and cauliflorin B

Objective To study the application effect of clinical pathway teaching in respiratory medicine. Methods Seventy clinical medical students of our department during 2007 to 2009 were randomly divided into two groups. The control group (35 students) was treated by traditional teaching ways, while the experimental group(35 students) was treated by clinical pathway teaching ways, with 5 to 6 students forming a small group. Teachers provided one copy of the CP version to each person in advance. Then progressive questions and discussions were conducted according to the diagnosis, dif-ferential diagnosis and treatment of CP. After its implementation for a certain time, students were co-mprehensively assessed by the practice examination and questionnaire and the statistical analysis was performed by using SPSS 17.0 version statistics software. Results The experimental group's total aver-age score was (90.00±4.00) points, while the control group's total average score was (76.00±7.20) points. There was significant difference between the two groups(P=0.001). The effect of these two kinds of teaching was evaluated and compared in stimulating interest in learning (P=0.002), improving the analytical ability(P=0.004),improving self-study ability(P=0.001), deepening the under-standing of the basic concepts(P=0.112), improving the innovation ability (P=0.005), improving the efficiency of learning(P=0.034), improving clinical comprehensive ability(P=0.016), and improving the ability of language expression(P=0.000), showing that teaching method of clinical pathway could significantly improve clinical teaching effect, and there was statistically significant difference between them. Conclusion Clinical pathway teaching has obvious advantages in cultivating students' diagnostic thinking and clin-ical ability to solve practical problems, and therefore it has better clinical teaching effect than the tra-ditional teaching method.

Objective:To analyze the applicability and feasibility of a cell-free DNA barcode-enabled single-molecule test (cfBEST) in non-invasive prenatal diagnosis of phenylketonuria.Methods:This study recruited four pregnant women who were prenatally diagnosed as heterozygous carriers of hot spot mutations in the PAH gene from pedigrees with phenylketonuria at the First Affiliated Hospital of Zhengzhou University from July to September 2019. The frequency of mutations in maternal plasma cell-free DNA and the fetuses' genotypes were analyzed by cfBEST. Nested polymerase chain reaction primers were designed to amplify the mutation sites in each pedigree. The results of cfBEST were compared with those of invasive prenatal diagnosis. Descriptive analysis was used for data analysis. Results:In pedigree 1, the frequency of c.603T>G and c.842+2T>A mutations in maternal plasma cell-free DNA were 48.40% (291/601) and 9.70% (61/628), which was detected by cfBEST. The fetus was diagnosed with phenylketonuria with two heterozygous mutations. In pedigree 2, the frequency of c.1238G>C and c.842+2T>A mutations in maternal plasma cell-free DNA was 43.70% (786/1 798) and 0% (0/1 550), respectively. Both mutations were wild-type, and the fetus was neither phenylketonuria nor a carrier. In pedigree 3, the frequency of c.1045T>G and c.728G>A mutations in maternal plasma cell-free DNA was 44.00% (930/2 112) and 0% (0/705), respectively, suggesting that both mutations in the fetus were wild-type, and the fetus was neither phenylketonuria nor a carrier. In pedigree 4, the frequency of c.755G>A and c.728G>A mutations were 45.40% (743/1 637) and 4.50% (28/849), respectively, which indicated that the former was wild-type, and the latter was heterozygous; namely the fetus was a carrier of phenylketonuria. The results of cfBEST were consistent with those of invasive prenatal diagnosis. Three pedigrees (Pedigree 2, 3 and 4) continued the pregnancy to full-term, and the phenylalanine levels in the neonates were all below 120 μmol/L. No abnormalities were reported in those three infants during follow-ups at one, three, and six months after birth.Conclusions:The cfBEST could be used for non-invasive prenatal diagnosis of phenylketonuria caused by PAH gene mutation, but further studies with a larger sample size are needed.

In order to verify whether p-nitrophenylalanine-containing BAFF vaccine can be used as a candidate molecule for the treatment of autoimmune diseases with BAFF over-expression,a soluble mutant of B cell activating factor belonging to the TNF Family (smBAFF) and its pNO2Phe mutant(pNO2Phe65smBAFF),which site specific incorporated pNO2Phe at position 65 of smBAFF,were expressed and purified.In order to evaluate the feasibility of using pNO2Phe65 smBAFF to treat BAFF-over-expressed autoimmune diseases,we investigate its Lymphocyte-stimulating capacity,immunogenicity and inhibitory effect of serum on biological activity of natural BAFF.The pharmacological activity of pNO2Phe65 smBAFF was evaluated using a cGVHD(graft-versus-host disease) induced SLE mouse model.Results indicated that pNO2Phe65 smBAFF,could bind to mouse lymphocytes but could not promote the proliferation of mouse lymphocytes.Moreover,the incorporation of pNO2Phe significantly increased the immunogenicity and induced cross-antibody,which can inhibit the biological activity of natural BAFF.In cGVHD induced SLE mouse model,pNO2Phe65 smBAFF can significantly reduce the symptoms of the disease and play a therapeutic role.Therefore,pNO2Phe65 smBAFF can be used as a candidate molecule for the treatment of autoimmune diseases with BAFF over-expression.

BACKGROUND: Endothelium-dependent vasodilatation (EDD) has been assessed by intracoronary acetylcholine infusion and digital subtraction angiography. However, its applications in continuous observing on early onset, progression and outcomes of clinical interventions of the disease are limited due to the potential trauma of the method. OBJECTIVE: This study was designed to assess the changes in brachial EDD in patients with hyperlipidemia using high-resolution ultrasound and to compare the outcomes of the patients to those of the healthy individuals. DESIGN: Case-control study SETTING: This study was carried out at the Cardiology Department and the Ultrasound Department of Xinyang CentralHospital PARTICIPANTS: Sixty patients with hyperlipidemia, 37 males and 23females, with an age ranging from 36 to 75 years old, were selected at the Cardiology Department of Xinyang Central Hospital from May 2001 to March 2002. They were divided into 3 groups according to abnormalities in blood lipid, as hypercholesterolemia group (HC, 20 cases),hypertriacylglycerolemia group (HTG, 20 cases) and mixed hyperlipidemia group (HLP,20 cases). Twenty voluntary individuals with normal physical examination findings were allocated into control group, 12 males and 8 females. Informed consents were obtained from all the eligible. METHODS: The brachial artery inner diameter and the changes in blood flow of the brachial artery were assessed using high-resolution ultrasound.After the patients having a rest for more than 10 minutes, the inner diameter (D0) and the baseline blood flow of their brachial artery were measured. The blood pressure cuff was placed under the elbow with a pressure at 300 mm Hg, lasting for 4 to 5 minutes and followed with a sudden deflation. 15 s after deflation, the reactive brachial artery inner diameter (D1) and the blood flow of the brachial artery were detected. The patients took another rest for more than 15 minutes and after the artery restored to a normal condition,the patients took nitroglycerol sublingually at a dose of 400 μg. Three to four minutes later, the brachial artery inner diameter (D2) and the blood flow of the brachial artery were detected. The increasing rate of the inner diameter of brachial artery were calculated, the increasing rate of reactive brachial artery inner diameter (%)= (D1-D0)/D0×100% and the increasing rate of post-treatment brachial artery inner diameter (%)=(D2-D0)/D0 ×100%.Meanwhile, the increasing rate of reactive blood flow (%)=(reactive blood flow-baseline blood flow)/ baseline blood flow (100% and the increasing rate of post-treatment blood flow (%)=(the post-treatment blood flow-baseline blood flow)/baseline blood flow×100%. MAIN OUTCOME MEASURES:In three hyperlipidemia groups and the control group, the inner diameter of the brachial artery were measured under resting conditions, in reactive congestion status and after sublingual nitroglycerol treatment respectively using high-resolution ultrasound. RESULTS: All the 60 patients with hyperlipidemia and 20 healthy controls entered the analysis procedure. In HC group, HTG group and HLP group, the increasing rate of reactive brachial-artery inner diameter were significantly lower than that in control group [(5.7±3.2)%, (5.4±3.0)%,(3.8±2.4)%, (11.3±3.1)%,P ＜ 0.05], which was most significant in HLP group. The increasing rate of both the reactive and the post-treatment blood flow in three experimental groups and the increasing rate of the post-treatment brachial-artery inner diameter were not significant (P ＞ 0.05). CONCLUSION:EDD were impaired significantly in patients with hyperlipidemia. High-resolution ultrasound is capable of assessing EDD accurately and reliably.

Objective: To determine the frequency, common chromosomal karyotypes and breakpoints, and involved regions among carriers of reciprocal translocations from Henan Province, and to explore the influence of common breakpoint regions on pregnancy and fetal development. Methods: For 586 carriers of reciprocal translocations, the above features were retrospectively analyzed. Results: The 586 reciprocal translocations were identified among 62 477 subjects, which yielded a frequency of 0.94%. Among these, 572 (0.92%) had abnormal fertility, and 14 (0.02%) had a history of abnormal fetal development. Statistical analysis showed that chromosomes 1, 4, 7 and 11 were most frequently involved, with t(11; 22)(q25; q13) being the most common type of translocation. In total 437 breakpoint regions were identified, with 11q23, 22q13 and 1p36 being most frequently involved, which resulted in infertility, abortion, embryo death, congenital malformation, development delay, mental retardation or a normal phenotype. Conclusion Above results indicated a 0.92% carrier rate for reciprocal chromosomal translocations in Henan. The location of breakpoint regions may affect the pregnancy and/or fetal development. Discovery of such regions may enable more accurate genetic, reproductive and developmental counseling for carriers, and provide reference for delineation of function and pathogenetic mechanism of the relevant genes.

OBJECTIVE: To explore cytogenetic characteristics and fertility of carriers of complex chromosome rearrangements (CCR) from Henan region. METHODS: G-banded karyotyping analysis was carried out on peripheral blood lymphocytes derived from 160 601 patients with reproductive abnormalities. Relevant literature was retrieved from domestic and overseas databases. Cytogenetic characteristics and clinical data of CCR carriers were analyzed. RESULTS: Twenty-seven CCR carriers were identified among the 160 601 patients. In addition, 6 cases were identified from the database research. Among the 33 CCR carriers, there were 17 three- and four-way exchange cases (51.5%), 10 double two-way exchange cases (30.3%), and 6 unusual CCRs (18.2%). Infertility was noted in 14 (42.4%) of the CCR carries. A total of 38 pregnancies were achieved in the remaining 19 cases (57.6%), among which spontaneous abortions or embryonic losses have occurred in 89.5% (34/38), multiple congenital abnormalities have occurred in 7.9% (3/38), while phenotypically normal offspring have occurred in 2.6% (1/38). Chromosomes 1, 11, 2, 4, 5 and 12 were more frequently involved, with their breakpoints occurred more than 3 times at 1p22, 11q23, 12p13 and 22q11. CONCLUSION CCR carriers are at a higher risk for abnormal pregnancies. Even for those with normal pregnancy, prenatal diagnosis should be provided. Chromosomes and their breakpoints involved in CCR may affect the fertility of CCR carriers. Analyzing the types of CCR and involved chromosomes and breakpoints can facilitate genetic counseling for CCR carriers.
Chromosome Aberrations ; Cytogenetic Analysis ; Female ; Fertility ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Pregnancy ; Translocation, Genetic

We report the use of droplet digital polymerase chain reaction (ddPCR) in non-invasive prenatal test fetal Charcot-Marie-Tooth disease (CMT) caused by MFN2 gene mutation. The proband, namely the husband, was found with heterozygous mutation of c.919A>G(p.K307E) in the MFN2 gene, which was diagnosed as CMT type 2A2A at a local hospital. The proband's wife underwent genetic counseling after conception at the First Affiliated Hospital of Zhengzhou University. Peripheral blood obtained from the pregnant woman was analyzed by ddPCR at eight gestational weeks, which found the fetus to carry a paternal pathogenic gene mutation. Sanger sequencing for the chorionic sample at 11 gestational weeks further verified that the fetus was a c.919G>A(p.K307E) heterozygous mutation carrier, the same as the proband. ddPCR could be applied to cell-free fetal DNA to detect the paternal pathogenic gene mutation in the non-invasive prenatal test.