Objective To analyze chemical constituents of Xionggui Decotion by rapid‐resolution (high performance) liquid chromatography‐time of flight mass spectrometry (HPLC‐TOF‐MS) .Methods A Shelloseido column(100 mm × 3 .0 mm ,3 μm)was used to separate .The mobile phase consisted of water containing 0 .1% methane acid and acetonitrile was used as gradient elute .The flow rate was 0 .4 ml/min .TOF‐MS was applied for qualitative analysis under positive ion mode . Results Under LC/MS condition ,47 major constituents in Xionggui Decotion were identified by time of flight mass spectrome‐try and structure‐relevant fragment ions .Conclusion A simple and reliable method using HPLC‐TOF‐MS was established to i‐dentify the chemical constituents of Xionggui Decotion .

Objective To explore the clinical effect of mycobacterium tuberculosis determination by culture and microscopy assay from sputum specimens of pulmonary tuberculosis patients.Methods The sputum specimens were dyed with the Ziehl-Neelsen anti-acid dyeing direct microscopy and examined by BACTEC-TB 960 culture system.Results The positive rate of culture and microscopy were 36.6% and 24.6%,the difference in the positive rate between the two methods was significant(P

Objective To investigate the effect of genistein on the expression of Bax and Bcl-2 protein in PC12 cells transfected with mutant APP695 Type.Methods PC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid,and then divided into normal control group,Unladen group,APP695 transfected group,GST treatment group(5 μmol/L,15 μmol/L).The Bax and Bcl-2 protein expression in PC12 cells was measured by Immunocytochemistry SABC and Western-blot.Results Immunocyto-chemistry SABC showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was strongly increased((78.02 ± 1.26) ％,P＜0.01)) and the expression of bcl-2 protein was very weak oppositely ((15.40 ± 0.72) ％,P＜0.01)).Compared with APP695 transfected group,the expression of bax protein in cells of 15umol/L GST treatment group was significantly decreased((22.35 ± 0.49) ％,P＜0.01))and the expression of bcl-2 protein in these cells was strongly increased ((68.11 ± 0.24) ％,P ＜0.01)).Western-Blot showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was markedly increased and the expression of Bcl-2 protein was significantly decreased.Compared with the APP695 transfected group,the expression of Bax protein was markedly decreased and the expression of Bcl-2 protein was significantly increased in the cells of the GST treatment groups.Conclusion GST can reduce the expression of bax protein and the increased expression of Bcl-2 protein has protective effect on PC12 cells transfected with APP696MT.

Objective To investigate current status of cognitive behavior on chronic obstructive pulmonary disease (COPD) to provide evidence for targeted intervention.Methods A questionnaire survey on COPD was conducted at 12 settings of 4 prefectures that were sampled from Ningxia Province using multistage stratified cluster random sampling method,and 4268 residents at least 40 years old were interviewed.Multivariate linear regression method was used for data analysis.Results A total of 4056 valid questionnaires were returned from 4200 participants with an effective response rate of 96.57％.Only 6.51％(264/4056) had an idea of COPD,13.88％ (563/4056) regarded cigarette smoking as a risk factor of chronic bronchitis and emphysema,and 6.39％ (259/4056) knew pulmonary function tests.Home income,living area,education level and ethnicity were main factors influencing COPD awareness.The percentage of current cigarette smoking was 28.80％ (1168/4056) with 20.40％ in Hui (Muslim) ethnic group and 34.63％ in Han ethnic group (P ＜ 0.05 ) ; smoking index and severe smokers were not significantly different between the two groups ( P ＞ 0.05 ).Conclusion Our data suggest a considerably poor knowledge and prevention awareness of COPD in Ningxia Province.In spite of relatively lower cigarette smoking rate,the problem that Hui (Muslim) ethnic people are lack of COPD awareness should not to be neglected.

Objective To study the feasibility of using heat and moisture exchangers (HME)as an alternative to heated humidifiers (HH) in patients undergoing mechanical ventilation. Methods 266 pa-tients with mechanical ventilation admitted to our ICU over the recent 3 years were allocated to the experi-mental group (humidification with a heat and moisture exchanger) and the control group (with heated hu-midifier), and the effect of humidification, the reserved time of artificial airway, the time on mechanical yen-tilation, the time of stay in ICU, the ineidenee of ventilator-associated pneumonia (VAP) and the mor-tality rate were comparatively studied and analyzed. Results Significant differences were found between the experimental and the control group in effect of humidification, insufficiency of humidification or excessive hu-midification, airway spasm and time on mechanical ventilation and time of stay in ICU. The incidence of VAP in the control group was significantly higher than that in the experimental group. There were no significant dif-ference between the two groups in the reserved time of artificial airway and the mortality rate. There were no accident of humidification occurred in the experimental group while there were one case complicated with air-way burn and 11 eases complicated with choking with water in the control group. Conclusions We conclude that HH can be replaced by HME on mechanical ventilation while disease evolution and effect of humidification should be monitored closely and keep HME unobstructed.

Objective To investigate the effect of phosphoinositide-3-kinase inhibitor LY294002 on the differentiation of human embryonic stem cells (HESC) into more mature insulin-producing cells. Methods HESCs were induced to differentiate into insulin-producing cells through five stages. Nicotinamide and B27 (group B27), nicotinamide and LY294002 (group LY) were used to induce the nesting positive cells into mature insulin-producing cells. The morphological change of each stage was observed under microscope , and expressions of insulin, c-peptide, somatostatin and glucagon were identified by immunofluorescence staining. Results After 14 days in stage 5 , there was no significant difference in rate of insulin positive cells between group LY and group B27 (P﹥0.05), but rates of somatostatin and glucagon positive cells in group LY were lower than those in group B27(P﹤0.05). Furthermore, the co-stained rate of somatostatin and insulin in group LY was also lower than that in group B27 (P﹤0.05). Conclusion HESCs can be induced to differentiate into more mature insulin-producing cells by phosphoinositide-3-kinase inhibitor LY294002 in serum-free culture medium.

Objective To evaluate the immunogenicity of a Haemophilus influenzae type b capsular-tetanus toxoid(Hib-TT) conjugate vaccine produced by Lanzhou Institute of Biological products(LIBP).Methods In an open-controlled,randomized trial,the eligible and consented 6-59 months-old young children injected 2 or 1 times 1 month apart with Hib-TT conjugate vaccine,the 3-5 months-old infants received 3 injections 1 month apart for primary immunization with Hib-TT or a licensed international Hib-TT conjugate vaccine as the control vaccine,and the boosting dose of two 3-5 months-old groups was injected at the 15-17months-old.The serum anti-Hib PRP IgG GMC in both groups after primary and boosting vaccination was measured by ELISA,the percentage of geometric mean concentration (GMC) ≥ 0.15 μg/ml and ≥ 1.0μg/ml was calculated,respectively.Results The Hib-TT conjugate vaccine produced in LIBP elicited satisfactory IgG antibody response in 3-59 months-old young children,the serum IgG GMC of anti-Hib PRP were 14.52 μg/ml(95％ CI:12.31-17.14)in 3-5 months-old,14.04 μg/ml(95％ CI:12.40-15.90) in 6-11 months-old.the ratios of IgG antibody concentration ≥ 1.0 μg/ml were 96.90％ (95％ CI:92.50-99.20) in study vaccine group and 98.55％ (95％ CI:92.20-99.90) in the control vaccine after 3 doses,respectively.100％ of the 6-11 months-old young children who injected 2 times with the Hib-TT conjugate vaccine had IgG antibody concentration ≥ 1.0 μg/ml (95％ CI:95.94-100.00),91.35％ (95％ CI:86.13-99.48) of recipients in 12-59 months-old young children induced the IgG antibody concentration ≥ 1.0 μg/ml after a single dose.The serum IgG antibody GMC in recipients who received the study or and control vaccines increased from 6.27 μg/ml (95 ％ CI:5.28-7.48) and 5.57 μg/ml (95 ％ CI:4.45-6.97)at pre-boosting injections to 63.14 μg/ml(95％ CI:52.14-76.47) and 73.48 μg/ml (95％ CI:57.37-94.11) one month after boosting injection,respectively.The percentage of IgG antibody concentration ≥ 1.0 μg/ml increased from 76.35％ and 79.55％ of pre-boosting to 100％ in the two groups after booting dose.Although the serum IgG GMC in two groups appeared to decline markedly,it remained at a relatively high levels of 25.02 μg/ml (95％ CI:20.51-30.48) in the study vaccine and 23.64 μg/ml (95％ CI:18.40-30.43) in the control vaccine,and all of the recipients in both groups remained 100.0％ of IgG antibody concentration ≥ 1.0 μg/ml.Conclusion The study vaccine elicited a protective immune response and induced the IgG antibody concentration which indicated long-term protection of anti-Hib PRP in 3 to 59months-old infants and young children.

Objectives To evaluate the safety and immunogenicity of Haemophilus influenzae type b capsular-tetanus toxoid(Hib-TT) conjugate vaccine.Methods In an open-controlled,randomized trial,the eligible and consented infants of 3 to 5 months-old received 3 doses of Hib-TT or a licensed Hib-TT conjugate vaccine(Anerbao) as the control vaccine to evaluate safety; The serum anti-Hib PRP IgG antibody mean geometric concentration (GMC) in both groups after primary and boosting vaccination were measured by ELISA.Results No apparent difference in the frequency of total adverse reactions observed between two groups (study vaccine 23.85％ vs.comparator 31.40％) (x2=0.5,P＞0.05).The mild and severe fever reaction of both vaccines was 3.67％ and 4.48％ respectively,with no significant difference.The local reactions including erythema,swelling and induration reported was 1.22％ in study vaccine group.After 3 injections,the serum anti-Hib PRP IgG antibody GMC was 6.6686 μg/ml in study group and 7.5346 μg/ml in control group,with no significant difference in both of the antibody GMC (x2 =0.147,P=0.702).After one dose boosting injection,the serum antibody GMC in study group increased from 2.6396 μg/ml of preboosting to 6.2044 μg/ml of post-boosting.Conclusion The Hib-TT conjugate vaccine is proved to be safe in 3-5 months-old infants.The primary immune schedule for 3-5 months-old infants of 3 injections 1 month apart with the Hib-TT conjugate vaccine could induce IgG antibody response against Hib PRP with the GMC of long-term protection concentration in serum.A boosting dose after primary vaccination elicited obviously immunological memory.

OBJECTIVETo assess the association of PEX10 gene and 1p36 copy number variations in 1p36 region with concurrent epilepsy through analyzing 3 cases.

METHODSThe karyotypes of 3 patients were determined by high resolution chromosome banding, multiplex ligation dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH) combined with single nucleotide polymorphism array (SNP) technology. Real-time PCR was carried out to determine the mRNA levels of PEX10 gene in peripheral blood of the patients.

RESULTSNo abnormality was found upon high resolution karyotyping. MLPA analysis showed that all of the 3 patients had a copy number variation of subtelomeric region in the short arm of chromosome 1, which was confirmed by FISH and SNP chip analyses. Case 1 and case 2 both had an epilepsy phenotype, and their copy number variations have encompassed the PEX10 gene. On the other hand, case 3 has absent epilepsy, and its PEX10 gene copy number was normal. Family investigation confirmed that the chromosome abnormalities in all of the 3 cases were of de novo type. Compared with healthy controls, real-time PCR showed that mRNA of the PEX10 gene was increased in case 1 but decreased in case 2.

CONCLUSIONThe abnormal expression of PEX10 gene resulting from copy number variations of 1p36 region may be associated with the epilepsy phenotype.

Child ; Chromosomes, Human, Pair 1 ; DNA Copy Number Variations ; Epilepsy ; genetics ; Female ; Humans ; Peroxins ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Receptors, Cytoplasmic and Nuclear ; genetics

OBJECTIVETo determine the molecular etiology for a muscular dystrophy pedigree with target region sequencing platform using hereditary myopathy capture array.

METHODSSpecific gene testing was performed based on the clinical diagnosis. Since no pathogenic mutation was found, target region sequencing with hereditary myopathy capture array combined with Sanger sequencing and bioinformatics analysis were employed in turn. PolyPhen and NCBI were used to evaluate the pathogenicity of identified mutation and conservation of the gene.

RESULTSTarget region sequencing indicated the proband has carried a heterozygous c.3353 A>C mutation of COL6A3 gene, which was confirmed by Sanger-sequencing in 4 affected individuals from the family. The same mutation was not detected in healthy members of the pedigree. Bioinformatics analysis suggested that the mutation has caused a highly pathogenic amino acid substitution from Histidine to Proline. The affected patients featured normal intelligence with mild myogenic damage by muscle biopsy, slightly increased serum creatine kinase and slow disease progression, which was consistent with Bethlem myopathy.

CONCLUSIONTarget region sequencing is an effective and efficient method for genetic testing. The heterozygous c.3353A>C mutation in exon 8 of the COL6A3 gene probably underlies the Bethlem myopathy with autosomal dominant inheritance.

Adult ; Amino Acid Sequence ; Amino Acid Substitution ; Base Sequence ; Collagen Type VI ; genetics ; Contracture ; genetics ; Exons ; Female ; Heterozygote ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Muscular Dystrophies ; congenital ; genetics ; Mutation, Missense ; Pedigree ; Young Adult