Objective To establish the determination method of Astragaloside Ⅳ in Shenqi-wuweizi tablet. Methods A high performance liquid chromatography method with evaporate light scattering detection was adopted. The analytes were successfully separated with Agilent-C18 column using acetonitrile and water (30:70) as mobile phase at 1.0ml/min. The parameter of ELSD was set at a evaporation tube temperature of 90℃ and nebulization at 50℃. The column temperature was set at 35℃. Results The linear range of Astragaloside Ⅳ was 41.68～521μg/ml, and the calibration curve was lnA=1.5461lnC+5.9564 with a coefficient of 0.9995 (n=6). The average recovery rate was 100.42%, with a relative standard deviation of 1.74%. Conclusion The method was simple, rapid, sensitive and accurate. Besides, it showed good repeatability as well as specificity, suitable for the quality control of Shenqi-wuweizi tablet.

Objective To analyze the changes of PAIgG, CD62P, CD4+CD25+ Foxp3+Tr,and IL-18 before and after treatment in peripheral blood of children with acute idiopathic thrombocytopenic purpura(ITP) and investigate the function of these factors in the pathogenesis of ITP.Methods Forty-one cases of acute ITP children were divided into the effective group(35cases) and the ineffective group (6cases) according to the clinical treatment. To detect PAIgG,CD62P,and the number of Tr cells by using flow cytometry ,IL-18 plasma levels by ELISA assay,and analyze the variations of these indicators before and after treatment in children with acute ITP. Results In the effective treatment group, PAIgG, CD62P before treatment were 53.05%,(14.18±5.04 )%, which were significantly higher than that after treatment [18.62%, ( 8.36±1.95 )%] and control group[5.26%,(2.65±0.59) %,all P<0.01],and PAIgG,CD62P after treatment were also higher than that in control group [all P<0.05].IL-18,CD4 + T lymphocytes, Tr/CD4+T-lymphocyte ratios before treatment [415.47 ±38.92 ) ng/L,( 25.64 ± 5.81 )%,( 2.67 ± 0.14 )%]were significantly lower than that after treatment [(512.85±42. 17)ng/L,(35.08±6.07)% ,(4.76±0.58)%] and control group[(506. 39±32.28) ng/L,(35.32±2.27)% ,(5.37 ±0.69)% ,all P<0.01]. IL-18, CD4 +T lymphocytes, Tr/CD4 +T-lymphocyte ratios after treatmenthad no statistically significant difference compared with control group( all P<0.05 ). In ineffective group, the test results of PAIgG, CD62P, IL-18, CD4 +T lymphocytes, Tr/CD4+ T-lymphocyte ratios showed no significant change before and after treatment( all P<0.05 ).IL-18 had negative correlations with PAIgG,CD62P respectively before and after treatment(all P<0.05 ). Tr cells / CD4 + T had negative correlations with PAIgG,CD62P respectively (all P<0.05). Conclusions The amount of Tr, IL-18 were reduced, while CD62P and PAIgG increased in peripheral blood of children with acute ITP. IL-18, Tr , CD62P and PAIgG play important roles in the pathogenesis of acute ITP.

OBJECTIVE To investigate deep fungal hospital infection and methods of its decreasing. METHODS Case histories of near 3 years of hospitalized patients from Jan 2002 to Dec 2004 were analyzed according to Diagnosis Standard of Nosocomical Infection under the items,such as the patients,age,underlying disease,sample,strain,and species distribution. RESULTS There were 360 fungus strains belonged to 15 species in all samples;the patients age was 14-94 with 20 kinds of various underlying diseases;the fungi included Candida albicans,and C.tropicalis,accounted for 61.1% and 17.2%,respectively;the samples were sputum,urine,vaginal secretion,BLA,throat swab and pus,which were 42.2%,21.1%,13.9%,8.4%,5.8%,and 3.6%,respectively;the respiratory department,ICU,and urology department were mainly involved. CONCLUSIONS To prevent fungal infection, measures such as hospital′s environment and management including reasonable use of antibiotics play a great role in monitoring hospital fungal infection and its epidemiology.

Background and purpose: Multidrug resistance of tumor cells is the main factor for the failure of chemotherapy. It is found that the apigenin has the anti-tumor effect, but its role in multidrug resistant cells was rarely reported. This study aimed to investigate the effect of apigenin on multidrug resistant breast cancer cell line MCF-7/ADR, and to explore the role of apigenin in reversing multidrug resistance. Methods: The MCF-7/ADR cells were cultured with different concentrations of apigenin, and the same cells were cultured with ADR in the control group. Thecell proliferation was detected by MTT, the cell cycle distribution was detected by PI, and the cell apoptosis was detect-ed by Annexin V/PI. The drug sensitivity in vitro was detected by the method of MTT, and the drug retention rate was detected by rhodamine 123 accumulation. The expression of P-gp protein was measured by Western blot, the RT-PCR method was used to detect the transcription of multidrug resistance gene MDR1. Results: The MCF-7/ADR cell prolif-eration was inhibited by the apigenin, the cell cycle progression was blocked by the apigenin, and the cell apoptosis was induced by the apigenin. There were significant differences between the apigenin group and the ADR group (P<0.05). The IC50 of ADR on MCF-7/ADR cell was (12.37±0.18) μg/mL with the apigenin effect, while the IC50 of ADR on MCF-7/ADR cell was (39.83±0.29) μg/mL without the apigenin effect (P<0.05). The reversal index was 3.22. The retention rate of rhodamine 123 in MCF-7/ADR cells in the apigenin group was higher than that in the ADR group. The MDR1 gene transcription level in MCF-7/ADR cells was higher than that in the MCF-7 cells, and the P-gp expression in MCF-7/ADR cells was higher than that in the MCF-7 cells. However, the level of MDR1 gene transcription and P-gp expression were down-regulated by the apigenin in the MCF-7/ADR cells. Conclusion: The apigenin had anti-MCF-7/ADR effect, and played the role of reversing multidrug resistance in the MCF-7/ADR cells. The mechanism may be related to down-regulation of the MDR1 gene transcription and the P-gp mediated drug e?ux function.

Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.
Animals ; Cations ; Cell Line ; DNA ; Genetic Vectors ; Kidney ; cytology ; Liposomes ; Magnetics ; Nanoparticles ; Swine ; Transfection

Objective To evaluate dynamic graciloplasty (DGP) for canine in situ anal reconstruction. Methods Seventeen dogs were randomly divided into experimental group and control group. In control group, on the stage 1, the gracilis muscle were dissected in situ, manometry performed intraoperatively; In experimental group, the gracilis muscle were dissected in situ and stimulated chronically starting 7 days postoperatively. On stage 2, abdominoperineal resection of anus and graciloplasty for anal reconstruction were performed in two groups. After 2 weeks recovery, manometry and muscular fatigue-resistant curve (MFRC) were observed while myostimulator is switched off and on. The muscle of neosphincter was biopsied. Results After chronic low frequency electrical stimulation (CLFS), the percentage of type Ⅰ fibers in the stimulation group was higher than the control group (P0.05), but functioning neosphincter pressure is different significantly (P

ObjectiveTo study the apoptosis of CD4 + T lymphocytes and the detection of immune function in patients with pulmonary tuberculosis and explore the clinical significance.MethodsThe mononuclear cells were separated from the blood of the tuberculosis patients or the healthy.The flow cytometry was used to measure the percentage of apoptotic CD4 + T lymphocytes,and the standard of T-lymphocyte subsets were detected by using SAP technology.The red cell immune function were determined by using yeast wreath way.Results The apoptosis rate of CD4 + T lymphocytes and CD8 + T lymphocyte was ( 15.882 ± 4.65 ) ％,and (27.69 ± 0.74) ％.The Immune complex positive rate ( 19.40 ± 0.58) ％ in patients with tuberculosis was significantly higher than those in controls ( P ＜ 0.01 ).C3b receptor positive rate in red blood cells was ( 17.73 ± 0.63 ) ％,( 46.48 ± 1.34 ) ％ in CD3 + T lymphocyte,( 28.12 ±0.69 ) ％ in CD4 + T lymphocyte,and the ratio of CD4/CD8 ( 1.0223 ± 0.09362) in the patients with tuberculosis was lower than the control group( P ＜ 0.01 ).There were certain relationships between the apoptosis rate of CD4 + T lymphocytes and the percentages of CD4 + T lymphocyte,the standard of T lymphocyte subsets and the red cell immune function.ConclusionsThe apoptosis rate of CD4 + T lympho,cytes in patients with tuberculosis were significantly higher than the healthy,which led to reducing the number of CD4 + T lymphocytes.There was positive correlation between red cell immunity and T-lymphocyte immunity,and the immunity in red cell and T- lymphocyte was lower than normal controls,which may be related to the immune pathogenesis of pulmonary tuberculosis.

OBJECTIVE: To explore the genetic etiology of a fetus with Cornelia de Lange syndrome type 1. METHODS: Clinical data of the fetus was collected. Genomic DNA was extracted from amniotic fluid and peripheral blood samples of the parents and subjected to low-depth copy number variant sequencing, whole exome sequencing (WES) and Sanger sequencing. Pathogenicity of the candidate variant was predicted based on the guidelines of American College of Medical Genetics and Genomics (ACMG). Minigene assay was used to assess the effect of the variant on mRNA splicing. RESULTS: WES revealed that the fetus has harbored a heterozygous c.5808+5gG>A variant in the intron of the NIPBL gene, which was predicted to affect the mRNA splicing. The same variant was not detected in either parent. The variant was not recorded in ExAC, 1000G and dbSNP databases. Comprehensive analysis showed that the variant was deleterious and may result in skipping of exon 31 during mRNA splicing. CONCLUSION The fetus was diagnosed with Cornelia de Lange syndrome type 1. Splicing variant identified by WES may be verified by minigene assay in vitro, which can provide more evidence for the prediction of its pathogenicity.
Cell Cycle Proteins/genetics* ; De Lange Syndrome/genetics* ; Female ; Fetus ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; RNA, Messenger

Objective:To study the clinical and laboratory characteristics of neonatal isolated sulfite oxidase deficiency (ISOD).Methods:An infant with neonatal ISOD admitted to our hospital was retrospectively analyzed. Using key words "isolated sulfite oxidase deficiency", "SUOX gene", "Infant, newborn", databases including CNKI, Wanfang database, National library and literature center of science and technology, China science paper online, PubMed, Web of Science and EMBASE (up to January 2021) were searched and literature review was conducted. The clinical manifestations, laboratory results, treatment and prognosis were analyzed.Results:Our patient was a full-term male infant with eye movement disorder, refractory seizures, feeding difficulties, increased muscle tone, developmental retardation and microcephaly. Urine sulfite paper-strip test was positive. Uric acid was normal. Whole exon sequencing (WES) revealed SUOX c.475G>T and c.1201A>G compound heterozygous mutations. Cranial MRI showed multiple encephalomalacia and brain atrophy at 5-month of age. The infant died at 8-month. In the literature review, a total of 29 articles and 32 cases of neonatal ISOD were found. 87.5% of the cases developed symptoms within 1-week after birth. All had convulsive seizures. Some of them had feeding difficulties, muscle tone changes, developmental retardation, microcephaly and ectopia lentis. Cranial imaging showed white matter cystic lesions and brain atrophy. Laboratory examination showed elevated urinary sulfite and S-sulfocysteine. Uric acid and xanthine/hypoxanthine were normal. Blood homocysteine was decreased. 23 cases received genetic testing and all of them had SUOX mutations. The treatment was mainly symptomatic relief and supportive treatment. During follow-up, 15 cases died, 13 cases survived and 4 cases were unknown. All the surviving children had drug-resistant convulsions and developmental retardation.Conclusions:Neonatal ISOD may present with refractory convulsions, feeding difficulties and developmental retardation. Cystic white matter changes and brain atrophy may be seen on cranial imaging. Elevated urinary sulfites, decreased blood homocysteine and normal uric acid are important clues for diagnosis. Genetic testing is helpful for early diagnosis.

OBJECTIVETo explore the expression of Bcl-2 mRNA and its effect on prognosis of patients with primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL).

METHODSReal time quantitative PCR was used to determine the expression of Bcl-2 mRNA in 40 PGI-DLBCL patients and 17 healthy controls. The association of Bcl-2 expression with clinicopathological features and prognosis of the patients was analyzed.

RESULTSThe expression level of Bcl-2 mRNA in PGI-DLBCL patients was 1.03 ± 0.93, significantly higher than that of the controls (0.41 ± 0.21) (P < 0.05). The expression of Bcl-2 mRNA in stage IIE-IV patients (1.28 ± 1.01) was significantly higher than that in the stage I-II2 patients (0.62 ± 0.61) (P < 0.05). The expression of Bcl-2 mRNA in patients with international prognostic index (IPI) score >2 (1.95 ± 1.27) was significantly higher than those with IPI score ≤ 2 (0.86 ± 0.75)(P < 0.05). The expression of Bcl-2 mRNA in patients with complete remission (CR) (0.71 ± 0.58) was significantly lower vs. 2.42 ± 0.91 in patients with no CR (P < 0.05). Univariate analysis indicated that β2-MG, IPI score>2, the Lugano staging, and Bcl-2 mRNA expression were associated with overall survival (OS) and progression-free survival (PFS) (P < 0.05). Multivariate analysis indicated that IPI score>2 was independently associated with OS (P < 0.05), and both IPI score >2 and Bcl-2 mRNA expression were independently associated with PFS (P < 0.05).

CONCLUSIONSThe expression of Bcl-2 mRNA in the tumor tissue of PGI-DLBCL patients is significantly higher than that in controls. PGI-DLBCL patients with higher expression of Bcl-2 have a poor chemotherapy response and inferior prognosis. IPI score >2 and higher expression of Bcl-2 mRNA are independent poor prognostic factors for PFS in PGI-DLBCL patients.

Disease-Free Survival ; Genes, bcl-2 ; Humans ; Lymphoma, B-Cell ; diagnosis ; genetics ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism