Objective To investigate the effect of ginsenoside-Rgl on paraurethral fascia fibroblastsmultiplication and the expression of proliferation cell nuclear antigen (PCNA) of stress urinary incontinence(SUI) women in vitro. Methods Specimens of human paraurethral fascia were obtained from 4 SUI womenduring tension-free vaginal tape (TVT) or tension-free vaginal tape-obturator (TVT-O) procedure.Fibroblasts were isolated and cultured by outgrowth technique. After reaching confluency fibroblasts weresubcultured every 5 days and cells after passage number 3 to 5 were used for assessment. The paraurethralfascia fibroblasts were treated with ginsenoside-Rgl at different concentrations (5, 10, 20 μmol/L) andfibroblnsts without Rgl were used as controL The multiplication conditions of paraurethral fascia fibroblastswere respectively detected by methyl thiazolyl tetrazolium (MTr) assay and the expression of PCNA byhistochemistry. Results ( 1 ) Compared with the control group, the growth rate of cells treated with differentconcentrations of Rgl after 72h [ (29±5 )%, (40±5 )%, (26±4)% respectively ] was significantly higher(P<0.01). (2)Compared with the control group, the stimulatory effect of Rgl on fibroblast growth wassignificant at 24 h (P<0.01), and peaked at 72 hi (29±5)% ,(40±5)%, (26±4)% respectively, P<0.01]. (3)Compared with the control group(28.77% ), there was a significant increase of PCNA-positivecells (P<0.01) after 48 h treatment with different concentrations of Rgl (49.24%, 83.48%, 54.50%respectively). Conclusion The results indicate that, at least in vitro, fibroblasts from paraurethral fasciataken from women suffering from SUI are able to proliferate after

Immune checkpoint inhibitors have become an important alternative for advanced non-small cell lung cancer (NSCLC) patients to surgery, chemotherapy, radiotherapy and targeted therapy. Monoclonal antibodies directed against immune checkpoint have shown better results in the application of first- or second-line treatment of NSCLC and for both squamous and non-squamous cell carcinoma patients, especially for those with positive PD-L1 tumor cells. Some comments will be made in present paper about the efficacy, biomarker, combined therapy and the resistant mechanism of immune checkpoint inhibitors.

Central nervous system ( CNS) injuries, such as cerebral ischemia, traumatic brain injury (TBI), and spinal cord injury (SCI), are often accompanied by complex pathological changes, and could lead to a variety of other neurological diseases.Neurons and glial cells are precisely regulated by many genes.MicroRNA ( miRNA ) are endogenous molecules discovered in recent years that regulate post transcriptional gene expression.They are highly expressed in the central nervous system and abnormal expressed under pathological conditions.They are involved in regulating variety of pathological processes after CNS injuries, and are CNS disease potential biomarkers.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into neuron-like cells in new environment after transplantation, and then to replace damaged cells for reconstructing neural circuit. OBJECTIVE: To establish the co-culture system between rat BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron-like cells in the co-culture system. METHODS: The neural cells obtained from Wistar rat fetal brain tissue and BMSCs gained from rat thighbone were co-cultured in Transwell culture plate. The morphological changes of BMSCs were observed and the special markers of neural cells in BMSCs were examined by immunofluorescence on the fifth day of the co-culture. The results were compared with control group which where BMSCs were alone cultured. RESULTS AND CONCLUSION: BMSCs in the neural cells co-culture system extended, were radial, connected each other. Neuron specific enolase immunoreactions showed positive results, showing neuron-like cells. The positive ratio of neuron specific enolase-positive cells was (33.0±10.5)%. However, BMSCs in the control group did not express neuron morphological character. Immunofluorescence exhibited that cells were negative for neuron specific enolase. These indicate that microenvironment provided by neurons improves the differentiation of BMSCs into neuron-like cells.

Objective:To determine the content of ferric oxide in calamine powder. Methods:Flame atomic absorption spectrom-etry ( FAAS) was used to determine the content of ferric oxide. The determination results were compared with those of the titration method. Results:There was a good linear relationship between the absorbance and the concentration of iron within the range of 0. 5-4μg·ml-1 , the relative standard deviation of the repeatability test was 1. 2%, the average recoveries were 100. 4% and 99. 4%, the limit of quantitation and detection was 0. 081 μg·ml-1 and 0. 024 μg·ml-1 , respectively. Conclusion:The method of FAAS is ac-curate and quick with good specificity and high sensitivity, which can be used for the determination of ferric oxide in calamine powder.

Objective:To develop a method for the determination of palladium in bendamustine hydrochloride by GFAAS. Meth-ods:The sample was destroyed by heat, and the content of palladium was determined by GFAAS with the detection wavelength of 247. 6 nm. Results:The absorbance and the content of palladium showed a good linear relationship within the range of 20-60 ng· ml-1(r=0. 998 4). The average recovery of palladium was 102. 9%(RSD=1. 7%, n=9). Conclusion: The method is sensitive and simple, which can be used for the determination of palladium in bendamustine hydrochloride.

Objective The protein kinase C ( PKC) is an essential signaling substance in the early protection of cells in pre-conditioning.This study was to investigate the role of PKC in the myocardial cells of the rat model of anoxia -reoxygenation (A-R) in-jury preconditioned with sufentanil . Methods Primary myocardial cells isolated and cultured for 5 days were allocated to a control , an A-R, a sufentanil preconditioning ( SF) , and a phorbol+sufentanil preconditioning ( PMA +SF) group.The A-R injury model was established with cultured myocardial cells from neonatal rats , which were preconditioned with sufentanil at the concentration of 0.000 3 μmol/L in the SF group or phorbol followed by sufentanil 10 minutes later in the PMA +SF group.Then the proliferation of the cells was detected , the optical density of the Cx 43 protein observed by immunofluorescence confocal technology , the apoptosis rate of the cells determined by flow cytometry, and the total Cx43 proteins calculated by Western blot. Results Cell proliferation was signifi-cantly increased in the A-R, SF, and PMA+SF groups as compared with the control (P<0.05), higher in the SF and PMA +SF than in the A-R group (0.498 0 ±0.0432 4 and 0.7240 ±0.1234 vs 0.325 8 ±0.023 5, P<0.05), and in the SF than in the PMA+SF group (P<0.05).Flow cytometry showed significantly increased rates of early , late, and total cell apoptosis in the A -R ([4.96 ±0.59], [18.77 ±0.92], and [23.73 ±0.51]%), SF ([5.86 ±0.38], [10.37 ±0.38], and [16.23 ±0.32]%), and PMA+SF group ([5.71 ±0.58], [5.54 ±0.43], [11.24 ±0.62]%) as compared with the control (P<0.05), remarkably lower in the SF and PMA +SF than in the A-R group (P<0.05), and the late and total cell apoptosis rates markedly lower in the SF than in the PMA+SF group (P<0.05). Conclusion Sufentanil has a protective effect and PKC agonists combined with sufentanil may add to the effect on myocardial cells in A -R injury.

Objective To explore the correlation of the gene polymorphism of the two single nucleotide polymorphisms(SNPs)rs1443434andrs925489onforkheadboxEl(FOXE1)withthehighnormalthyroidstimulating hormone ( TSH) level in Chinese Han population. Methods 1 400 subjects with normal serum TSH and thyroid peroxidase antibody(TPOAb) levels were included. According to TSH or TPOAb levels, the subjects were divided into high normal TSH group(H-TSH group,n=195) and normal TSH control group(TSH control group,n=1 205) or high normal TPOAb group ( H-TPOAb group, n=711 ) and low normal TPOAb group ( L-TPOAb group, n=689 ) , respectively. The genotypes on the two SNPs of all the subjects were performed by whole-genome genotyping chips. Results There were significant differences in rs925489 genotypic distributions and allele frequencies between H-TSH group and TSH control group(both P<0. 05). The genotype TT and allele T in H-TSH group were significantly higher than those in TSH control group(89. 75% vs 83. 15%, 94. 62% vs 91. 29%). The normal TSH levels were positively associated with rs925489 genotypic distributions after adjustment for sex, age, and high density lipoprotein cholesterol(P<0. 01). There were no significant differences in rs1443434 genotypic distributions and allele frequencies between two TSH groups or two TPOAb groups. Conclusion FOXE1 rs925489 gene polymorphism may be correlated with the high normal TSH level in Chinese Han population.

BACKGROUND:Pulmonary aspergilosis is a disease caused by pulmonary fungal infection. Its diagnosis and treatment is usualy delayed because of nonspecific clinical symptoms, physicial sign and imaging changes as wel as uncertainties of histological and bacterial findings. Therefore, it is necessary to establish mouse models of invasive pulmonary aspergilosis to investigate the underlying pathological mechanism and novel therapeutic methods. OBJECTIVE: To establish mouse models of invasive pulmonary aspergilosis, detect the expression ofγ-interferon, Tol-like receptor 2 and Tol-like receptor 4, and discuss the mechanism of action underlying invasive pulmonary aspergilosis. METHODS:Seventy-five female BALB/c mice of clean grade, aged 6-8 weeks, were randomly and evenly divided into five groups: blank control group (group A), immunosuppressive model group treated with high concentrations of Aspergilus fumigatus spore suspension (group B), normal infection group treated with high concentration of Aspergilus fumigatus spore suspension (group C), immunosuppressive model group treated with low concentration of Aspergilus fumigatus spore suspension (group D), normal infection group treated with low concentration of Aspergilus fumigatus spore suspension (group E). First, mice in the groups B and D were intraperitonealy injected with cyclophosphamide to establish immunosuppressive models. The mice in the groups D, E (108 cfu/mL) and groups B, C (109 cfu/mL) were treated with 12 mL Aspergilus fumigatus spore suspension through the use of nebulizer. Mice in the group A were treated identicaly with sterile PBS. At 1, 3, 5 days of infection, the pathological change of lung tissue was observed, the mass concentration of γ-interferon in bronchoalveolar lavage fluid and the expression levels of γ-interferon mRNA and Tol-like receptor 2 and Tol-like receptor 4 mRNA and protein in the lung tissue were determined. RESULTS AND CONCLUSION:Abscess, spores and very severe bleeding and congestion, widenened alveolar septum and tracheal epithelial cel shedding and necrosis were observed in the mouse lung tissue in the group B. At 5 days of infection, the mass concentration of γ-interferon in bronchoalveolar lavage fluid and the expression ofγ-interferon mRNA in the lung tissue in the group B were significantly decreased compared with the group A (P < 0.05). Tol-like receptor 2 expression was strongly positive in the group B. Tol-like receptor 2 expression in the group C was significantly lower than that in the group B (P< 0.05). Tol-like receptor 4 expression was positive in the groups B and C, and its expression in the group C was significantly greater than in the group B (P < 0.05). The expression of Tol-like receptor 2, 4 mRNA in the mouse lung tissue of group B was significantly increased at 1, 3, 5 days of infection (P < 0.05). These results suggest that atomizing high concentration of aspergilus fumigatus spore suspension to immunosuppressive mice can establish stable invasive pulmonary aspergilosis models with typical pathological features. The infection of aspergilus fumigatus can activate tol-like receptor 2, 4 at the same time, and the pathological mechanism is closely related to organism’s immune defense function.

Objective:To develop a quantitative HPLC method for the analysis of eight impurities in active pharmaceutical ingredi-ent (API) asenapine maleate. Methods:The substances were analyzed using an Inertsil ODS-3 C18 column (250 mm × 4. 6 mm, 5μm), and gradiently eluted by the mobile phase A of 0. 1% aqueous trifluoroacetic acid and mobile phase B of acetonitrile in a flow rate of 1. 0 ml·min-1 with the detection wavelength of 220 nm and the column temperature of 35℃. Results:Asenapine was separa-ted completely from the impurities. The calibration curve of asenapine was linear within the range of 0. 45-1 458 μg · ml-1 ( r =1. 000 0), and that the impurities was linear within the range of 0. 4-30. 0μg·ml-1(r>0. 999). The mean recovery of the impurities was 93. 1%-106. 7%(n=9). Conclusion:The method is simple,sensitive and reproducible with good specificity and reliability,which can be used in the quality control of asenapine maleate.