Statistic-based experimental design was used to optimize the culture conditions for Pseudomonas aeruginosa ME16 strain which is capable of utilizing methane.Effects of inoculum volume,temperature,methane content,and initial pH of media on cell growth were studied with the medium of mineral salt and methane gas,and the response surface methodology(RSM) was then used to optimize culture conditions.The optimum conditions were as follows:temperature of 29.4℃,inoculum volume of 1.8% and methane content of 25%.Under the optimized culture conditions,the ME16 production was increased by 0.8 times and its cultivation time was reduced.The removal of methane gas by ME16 was preliminary investigated,and the removal rate of methane gas reached 65.7%.The results indicated that the strain was capable of removing of methane.

OBJECTIVETo understand the characteristics of human calicivirus (HuCV) infection in infants with diarrhea in Guangzhou city and to study genotype of the virus.

METHODSThe authors collected fecal specimens from 22 children with acute nonbacterial gastroenteritis from November to December, 2001. HuCV was detected from the specimens by RT-PCR. The PCR products were cloned into the PMD18-T cloning vector and sequenced.

RESULTSHCV was detected from the specimens of 2 cases (9%, 2/22). The nucleotide sequence analysis revealed that the virus strains belonged to genotype 2 of Norwalk-like viruses.

CONCLUSIONHuCV is one of the pathogens causing diarrhea in infants and young children in Guangzhou area. HuCV infection occurred sporadically in autumn and winter.

Base Sequence ; Caliciviridae ; genetics ; Caliciviridae Infections ; complications ; virology ; China ; DNA, Viral ; chemistry ; genetics ; Diarrhea, Infantile ; etiology ; Dysentery ; etiology ; Feces ; virology ; Genotype ; Humans ; Infant ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo observe the serological and epidemiological efficacy of hemorrhagic fever renal syndrome (HFRS) vaccine in Zhejiang province.

METHODSImmunofluorescent antibody assay and Mcro-CPE method were used to test specific IgG antibody and the titer of neutralizing antibody.

RESULTSTwo weeks after the injection of the third dose, the sero-conversion rates by both immunofluorescent antibody test (IgG) and neutralization test were 100.0% (67/67) (95% CI: 96.3 - 100.0) and 44.4% (8/18)(95% CI: 22.0 - 69.0) with geometric mean titers (GMTs) 72.1 and 4.6 respectively. The rates of seroconversion of immunofluorescent antibody by immunofluorescence antibody assay (IFA) were 28.6%, 83.3%, 75.0%, 53.1%, 22.6%, 10.0% and 55.0% before reinforcement, two weeks, one year, one year and a half years, two years, three years and five years after reinforcement. The rates of neutralizing antibody seroconversion by the Mcro-CPE method were found as 14.8%, 55.6%, 35.0%, 31.3%, 26.0%, 10.0% and 50.0% respectively. We found some antibody dependent immunization enhancement phenomenon among the inoculated population, but further observation was needed.

CONCLUSIONHFRS vaccine was immunologically effective and the duration of serous antibody last long.

Adolescent ; Adult ; Antibodies, Viral ; blood ; China ; epidemiology ; Fluorescent Antibody Technique ; Hantaan virus ; immunology ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; prevention & control ; Humans ; Immunization Schedule ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Neutralization Tests ; Vaccination ; Viral Vaccines ; immunology

OBJECTIVETo study the "hot spot" of BRCA1/2 gene mutations in Chinese mainland breast cancer population.

METHODSThe known BRCA1/2 gene mutations in author's previous studies were reanalyzed by denaturing high performance liquid chromatography and DNA sequencing method in 177 patients with early onset breast cancer or affected relatives and 426 sporadic breast cancer patients from four breast cancer centers in China.

RESULTSThree cases were found with BRCA1 5589del8 mutation out of 247 hereditary-predisposing breast cancer patients (70 patients in previous study and 177 patients in current study) and 2 cases with BRCA1 5589del8 mutation out of 426 sporadic breast cancer patients. They had similar even same haplotype.

CONCLUSIONBRCA1 5589del8 mutation is likely to be the "founder mutation" in Chinese population, but it should be confirmed by further studies.

Adult ; Asian Continental Ancestry Group ; genetics ; BRCA1 Protein ; genetics ; Base Sequence ; Breast Neoplasms ; ethnology ; genetics ; China ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; Female ; Genetic Predisposition to Disease ; genetics ; Humans ; Mutation

OBJECTIVETo explore clinical and chest X-ray features of SARS in children to facilitate correct diagnosis.

METHODSClinical manifestations and chest X-ray findings in five children suffering from SARS admitted for treatment in the hospital between February and May, 2003 in Shenzhen area were analyzed. The diagnosis was confirmed by epidemiological, clinical, laboratory and radiological examinations. Among the 5 cases, 1 was a boy and the others were girls at the age of 4 to 13 years.

RESULTSOf the 5 SARS children, 3 presented a history of close contact with SARS patients. Fever was the initiative symptom, 4 had a body temperature of over 38 degrees C with the highest being 40 degrees C; fever sustained from 4 to 7 days with an average of 5.6 days. All the 5 cases developed nonproductive cough; on auscultation, both moist and dry rales could be heard in 3 out of the 5 cases. Mean total white count of peripheral blood was (2.96 - 6.9) x 10(9)/L, and was < 5.0 x 10(9)/L in 4 cases. SARS associated coronavirus specific RNA fragment was found positive by RT-PCR in 1 case; 1 case was positive for both IgM and IgG antibodies to the virus; 1 case was positive for only IgM antibody and another 2 cases were positive for only IgG antibody. IgG and IgM antibodies to Mycoplasma pneumoniae and Chlamydia pneumoniae as well as blood culture for bacteria were all negative. Findings on chest X-ray examination: 4 cases showed presence of patchy or macular opacities with cord-like shadows in unilateral lung plates while 1 case each showed ground-glass-like opacity and migratory changes; 1 case showed interstitial changes in the lungs in the form of irregular reticular lattice and cord-like shadows. Two cases received CT scanning and macular-patchy or spotty shadows were seen all over the lung. The shortest time for absorption of foci in the lungs was 7 days while the longest was 33 days with a mean of 15 +/- 6 days. None of the cases had any signs of fibrosis in the lungs. All the 5 cases were completely cured and discharged 7 to 40 days (mean 18 +/- 11 days) after admission.

CONCLUSIONCompared with adult cases with SARS, children with SARS had milder symptoms and signs. Presence of unilateral patchy shadow in lungs represented the main chest X-ray findings.

Adolescent ; Antibodies, Viral ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin G ; analysis ; Immunoglobulin M ; analysis ; Male ; Radiography, Thoracic ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; immunology ; Severe Acute Respiratory Syndrome ; diagnostic imaging ; pathology ; virology

Objective: To clarify the molecular basis of patients with Bartter syndrome type I and explore the therapeutic effect of trafficking-defective variations by chemical chaperone 4-Phenylbutyric acid(4-PBA). Methods: The clinical characteristics, laboratory findings and genetic data of 3 patients diagnosed with Bartter syndrome type I who were admitted to Department of Nephrology, Children's Hospital of Nanjing Medical University from 2017 to 2018 were retrospectively analyzed. Wild type and variant SLC12A1 gene constructs were transiently overexpressed in HEK293 cells. Western blotting was used to detect the expression levels of Na⁺-K⁺-2Cl^-cotransporter(NKCC2) protein. Immunofluorescent staining was applied to investigate the subcellular localization of NKCC2 protein. In addition, the effect of the chemical chaperone 4-PBA on the expression and localization of the SLC12A1 gene variants was investigated. Unpaired t test was used for statistical analysis of 4-PBA treatment. Results: All the 3 patients (2 males and 1 female), aged 3.0, 4.0 and 1.2 years, respectively. All patients had antenatal onset with polyhydramnios and were born prematurely. After birth, all patients presented with hypochlorine alkalosis accompanied by hypokalemia and hyponatremia. Sequencing analysis revealed that the 3 patients were homozygotes or compound heterozygotes for variants in the SLC12A1 gene. In HEK293 cells, the surface expression of NKCC2 in 3 variants (p.L463S, p.L479V, p.507-510del) are all lower than in wild type (0.718±0.039, 0.287±0.081, 0.025±0.156 vs. 1.001±0.028, t=5.92, 8.35, 30.49, all P<0.01). Moreover, the total protein expression of p.L479V and p.507-510del group were all lower than that in wild type group (0.630±0.032, 0.043±0.003 vs. 1.000±0.111, t=3.21, 8.65, all P<0.05). 4-PBA treatment increased the mature protein expression level of the p.L463S and p. L479V group in 4-PBA treatment group are all higher than the untreated group (0.459±0.018 vs. 1.123±0.024, 0.053±0.012 vs. 1.256±0.037, t=2.75, 18.35, all P<0.05). Cytoplasmic retention of the L479V and 507-510del variants were observed by immunofluorescent staining. 4-PBA treatment could rescue a number of NKCC2 L479V variants to the membrane. Conclusions: The 3 SLC12A1 variants cause expression or subcellular localization defects of the protein. The findings that plasma membrane expression and activity can be rescued by 4PBA might help to develop novel therapeutic strategy for Bartter syndrome type Ⅰ.
Bartter Syndrome/genetics* ; Child, Preschool ; Female ; HEK293 Cells ; Homozygote ; Humans ; Infant ; Male ; Pregnancy ; Retrospective Studies ; Solute Carrier Family 12, Member 1/genetics*

To study the improvement of powder flowability and hygroscopicity of traditional Chinese medicine extract by surface coating modification technology. The 1% hydrophobic silica nanoparticles were used as surface modifier, and andrographis extract powder was taken as a model drug. Three different techniques were used for coating model drugs, with angle of repose, compressibility, flat angle and cohesion as the comprehensive evaluation indexes for the powder flowability. The powder particle size and the size distribution were measured by Mastersizer 2000. FEI scanning electron microscope was used to observe the surface morphology and structure of the powder. The percentage of Si element on the powder surface was measured by energy dispersive spectrometer. The hygroscopicity of powder was determined by Chinese pharmacopoeia method. All of the three techniques can improve the flowability of powder extract. In particular, hygroscopicity of extract powder can also be improved by dispersion and then high-speed mixing, which can produce a higher percentage of Si element on the powder surface. The improvement principle may be correlated with a modifier adhered to the powder surface.

The purpose of this study was to investigate the effect of key physical properties of hawthorn leaf granule on its dissolution behavior. Hawthorn leaves extract was utilized as a model drug. The extract was mixed with microcrystalline cellulose or starch with the same ratio by using different methods. Appropriate amount of lubricant and disintegrating agent was added into part of the mixed powder, and then the granules were prepared by using extrusion granulation and high shear granulation. The granules dissolution behavior was evaluated by using equilibrium dissolution quantity and dissolution rate constant of the hypericin as the indicators. Then the effect of physical properties on dissolution behavior was analyzed through the stepwise regression analysis method. The equilibrium dissolution quantity of hypericin and adsorption heat constant in hawthorn leaves were positively correlated with the monolayer adsorption capacity and negatively correlated with the moisture absorption rate constant. The dissolution rate constants were decreased with the increase of Hausner rate, monolayer adsorption capacity and adsorption heat constant, and were increased with the increase of Carr index and specific surface area. Adsorption heat constant, monolayer adsorption capacity, moisture absorption rate constant, Carr index and specific surface area were the key physical properties of hawthorn leaf granule to affect its dissolution behavior.