AIM: To investigate the effect of fetal bovine serum (FBS) on the proliferation and differentiation of murine corneal epithelial cells in vitro.METHODS: Mouse corneal epithelial cells(MCEs) were cultured in serum-free low-Ca2+ medium(KSFM) and KSFM supplemented with 100mL/L FBS, respectively. Population doublings (PDs) were determined. The expressions of corneal epithelial cell markers p63, keratin 19 (K19) and involucrin were investigated by RT-PCR and Western blotting analyses. RESULTS: Cells in KSFM were stably subcultured over 25 passages; however, none of the cell lines could pass P3 in KSFM with FBS. In KSFM, the cells showed typical cobblestone appearance and expressed p63, K19 and involucrin. After medium was supplemented with FBS, cells became homogeneous, large and squamous. Furthermore, both RT-PCR and Western blotting analyses showed that the expression of involucrin was increased significantly.CONCLUSION: FBS has effects of inhibiting proliferation and triggering differentiation of MCEs.

OBJECTIVETo discuss the value of grey scale ultrasonography and color Doppler of bi-planar transrectal ultrasound on the diagnosis of hyperplasia prostate gland (HPG).

METHODSOne hundred and ninety-two patients with HPG and 50 control subjects underwent abdominal ultrasound and TRUS examinations. The width (W), depth (D) and length (L) of the prostates and transitional zones were respectively measured. Each transitional zone index (TZI), i.e., the ratio of the transitional zone volume to the total prostate volume was calculated. Meanwhile, we observed the blood flow distribution in the prostates, calculated the parameters of hemodynamics such as peak systolic velocity (PSV), resistance index (RI) and pulse index (PI), and analyzed their correlations with TZI.

RESULTSThe revealing rates of cystic formation, calcium calculus and ejaculatory cyst and PSV, RI of arterial vessels in HPG were all significantly higher in the HPG patients than in the control subjects, and the revealing rates of all the above pathological changes but calcium calculus by TRUS were significantly higher than by abdominal examination. TZI of HPG was positively correlated with PSV, RI and PI.

CONCLUSIONBi-planar TRUS can clearly display the normal and pathological structure of the prostate. And PSV, RI and PI may aid in the diagnosis of HPG and the assessment of the degree of prostatic proliferation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostate ; blood supply ; diagnostic imaging ; pathology ; Prostatic Hyperplasia ; diagnostic imaging ; pathology ; Regional Blood Flow ; Ultrasonography, Doppler, Color ; methods

Amyloidosis ; diagnosis ; diagnostic imaging ; Female ; Humans ; Lung Diseases ; diagnosis ; diagnostic imaging ; Middle Aged ; Pneumonia ; diagnosis ; diagnostic imaging ; Radiography

OBJECTIVETo discuss the treatment strategy of severe hand-foot-and-mouth disease (HFMD) cases, prevent the severe cases from progressing to fatal condition and enhance the survival rate of critically ill patients.

METHODSEighty HFMD cases were divided into four groups, A, B, C and D, according to the severity of patients' nervous system manifestation and other system involved. Different intensive care and treatments were used and the effect and outcome were analyzed for each group. All statistical analyses were performed by using SPSS software 13.0. One-way ANOVA and Chi-square test were used for data analysis.

RESULTSThe most common symptoms were continuous fever (69/70) and myoclonic jerk (67/70). The fewer the rashes were, the more severe the patient's condition was, heart rate >200/min, hypertension, increase of white blood cells in peripheral blood and hyperglycemia were common in patients with lesions in brain stem and pulmonary edema. There were no relations between patient's conditions and CSF white blood cells and CRP. CNS involvement was highly associated with EV71 infection. There were 69 cases in group A, B and C in total and all recovered. Of 11 patients in group D, 6 got complicated neurogenic pulmonary edema and circulatory failure, 2 cases died and 9 cases survived, 8 cases recovered without sequelae while one case had sequelae of mental retardation and dyscinesia.

CONCLUSIONAdministration of mannitol, methylprednisolone, IVIG and other supportive treatments in time and reasonably might have advantages in avoiding aggravation of the condition and enhancing the rate of successful rescue in patients with nervous system involvement.

Adolescent ; Child ; Child, Preschool ; Critical Care ; Female ; Hand, Foot and Mouth Disease ; diagnosis ; drug therapy ; mortality ; Humans ; Infant ; Intensive Care Units, Pediatric ; Male ; Nervous System ; virology ; Survival Rate

OBJECTIVETo study the chemical constituents of Gentiana veitchiorum.

METHODThe chemical constituents were isolated by chromatography and identified by spectral data.

RESULTFive glycosides, loganic acid (1), gentiopicroside (2), isoorientin 3'-methyl ether (3), isovitexin (4), isoorientin (5) were isolated and identified.

CONCLUSIONCompounds 1-5 were isolated from this plant for the first time.

Apigenin ; chemistry ; Chromatography, High Pressure Liquid ; Gentiana ; chemistry ; Glucosides ; chemistry ; Glycosides ; chemistry ; Iridoid Glucosides ; Iridoids ; chemistry

Objective To investigate the prevalence of Vibrio parahaemolyticus(VP)in oysters in Jinshan District, Shanghai and make assessment on the risks that may cause, providing the basis for prevention and control of foodborne disease. Methods Raw oyster samples with shells were randomly collected from markets, supermarkets and restaurants in Jinshan District from July to October in 2017. The content of VP in oysters was tested in accordance with the national standard methods. The semi-quantitative risk assessment for VP in oysters was made by Risk Ranger combining with the monitoring results of diet and health status of residents in Jinshan District of Shanghai in 2012-2013. Results The overall positive rate of VP in the 40 oyster samples was 80.0%(32/40). The positive rate of VP in oyster samples from farmer's markets was the highest, 85.7%(12/14), followed by those from restaurants and supermarkets. The relative risk for VP in raw oysters was 63. The probability of illness caused by VP in oysters per day per consumer of interest was 6.85×10^-4, and the total predicted patients annual incidence rate in this population was 1 247.8/10⁵. Conclusion The contamination of VP in seafood oysters in Jinshan District is serious. Eating raw oysters is at high risk; consumers are advised to reduce or avoid eating raw oysters, and processing food before eating is an effective method to decrease VP infection. Strengthening surveillance and management is imperative in this regard.

OBJECTIVE: The aim was to investigate the polymorphisms of D6S1043 and D12S391 loci among Han population and evaluate their values in paternity testing. MERTHODS: By using fluorescence dye-labeled primers and capillary electrophoresis, the allele frequencies of the two STR loci among 192 unrelated individuals were investigated. RESULTS: Twelve alleles were observed in both D6S1043 and D12S391 loci. The ranges of allele frequencies were from 0.0026 to 0.1719 and from 0.0026 to 0.2292, respectively. The discrimination power of D6S1043 and D12S391 were 0.9656 and 0.9510. The Average exclusion probability in paternity testing for duos were 0.573 and 0.510. The Average exclusion probability in paternity testing for trios were 0.731 and 0.679, respectively. The genotypes frequencies met Hardy-Weinberg equilibrium expectation. CONCLUSION The results show that D6S1043 and D12S391 have high values in forensic paternity testing.
Alleles ; China/ethnology* ; DNA Fingerprinting/methods* ; Electrophoresis, Capillary ; Forensic Medicine/methods* ; Gene Frequency ; Genetics, Population ; Humans ; Paternity ; Probability ; Tandem Repeat Sequences/genetics*

BACKGROUNDEndothelial progenitor cells (EPCs) derived from bone marrow may differentiate into endothelial cells and participate in endothelial repair. These cells can be mobilized into peripheral blood by cytokines, including granulocyte colony-stimulating factor (G-CSF). In the present study, we investigated the effects of G-CSF on neointimal formation and restenosis in a canine model of arterial balloon injury.

METHODSSixteen male beagle dogs were injected subcutaneously with 20 microg x kg(-1) x d(-1) recombinant human G-CSF (n = 8) or normal saline (n = 8) for 1 week. On the fifth day of treatment, the dogs underwent renal arterial angioplasty. At 8 weeks after arterial balloon injury, angiographic observations were made and injured arteries were processed for morphometric analysis of neointimal formation.

RESULTSPeripheral white blood cell counts were increased by 3.34-fold compared to baseline on the fifth day of administration of G-CSF. Angiographies revealed that one stenosis had occurred among the eight injured renal arteries from dogs treated with G-CSF, whereas all injured renal arteries from dogs treated with normal saline remained patent. The mean extent of stenosis among injured arteries was 18.3% +/- 17.9% in the G-CSF treated group compared to 12.5% +/- 7.6% in the saline treated control group (P = 0.10). G-CSF treatment slightly increased neointimal thickness (0.42 +/- 0.15 mm vs 0.25 +/- 0.06 mm, P = 0.08) with an intima to media ratio of 0.83 +/- 0.49 vs 0.54 +/- 0.18 (P = 0.11).

CONCLUSIONSG-CSF treatment does not attenuate neointimal hyperplasia and restenosis formation in a canine model of renal arterial injury, suggesting that the therapeutic strategy for preventing restenosis by stem cell mobilization should be investigated further.

Animals ; Dogs ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Hyperplasia ; Male ; Recombinant Proteins ; Renal Artery ; injuries ; pathology ; Tunica Intima ; pathology

OBJECTIVE: To explore the appropriate amount of template DNA for Sinofiler Kit. METHODS: The DNA samples with ideally genotyped results by Sinofiler Kit were detected by real-time quantitative PCR assay. RESULTS: It was shown that 1.29-1.51 ng of template DNA in 12.5 microL reaction volume was optimal for STR genotyping with Sinofiler Kit. CONCLUSION Real time quantitative PCR is an accurate and necessary technique for detection of appropriate amount of template DNA for different kits.
DNA/analysis* ; Forensic Medicine/methods* ; Hair/chemistry* ; Humans ; Microsatellite Repeats ; Reagent Kits, Diagnostic ; Reverse Transcriptase Polymerase Chain Reaction/methods* ; Templates, Genetic

OBJECTIVE: To explore the immune cytotoxic effect and the maximum non-effect dose of trichloroethylene( TCE) on Jurkat T cells in vitro. METHODS: i) Naive and activated Jurkat T cells were treated with different concentrations of TCE( 0. 10, 0. 50, 1. 00, 2. 00, 5. 00, 10. 00 mmol / L). Phorbol-12-myristate-13-acetate and ionomycin were used as agonist. No TCE was used in the control group and dimethyl sulfoxide( DMSO) was used as the solvent group. The morphology of Jurkat T cells was observed using a light microscope and the survival rate of Jurkat T cells was investigated using CCK-8 essay after cells were cultured for 24,48 and 72 hours. ii) Nave and activated Jurkat T cells were treated with different concentrations of TCE( 0. 00,0. 02,0. 20,2. 00 mmol / L). The apoptosis of cells was detected using flow cytometry and the level of interleukin-2( IL-2) in supernatant was detected using enzyme linked immunosorbent assay after cells were cultured for 24,48 and 72 hours. RESULTS: i) Cytotoxic effect was observed after cells were exposed to 10. 00 mmol / L TCE for 24 hours. Cells dispersed,cell volume diminished,cell membrane ruptured,cytoplasm condensed and increased outflow of intercellular organelles. The effect of interaction between exposure dose and exposure time was statistically significant on cell survival rate( P < 0. 01). Compared with the control and DMSO groups at the same time points,there were no significant differences in the 0. 10,0. 50,1. 00 and 2. 00 mmol / L TCE treatment groups in cell survival rates in three different time points( P > 0. 05),while the cell survival rates of 5. 00 and 10. 00 mmol / L TCE treatment groups were significantly decreased( P < 0. 01). ii) When TCE concentration was 0. 00-2. 00 mmol / L,there were no significant differences in the main effect of exposure dose and interactions of between exposure dose and cell type or exposure time on cell apoptosis rate( P > 0. 05). Compared with the same time points and groups of naive Jurkat T cells,the levels of IL-2 of activated Jurkat T cells were significantly increased( P < 0. 01). In the three different time points,the level of IL-2 of activated Jurkat T cells increased in accordance with the TCE exposure dose,showing a dose-effect relationship( P < 0. 01). The level of IL-2 of activated Jurkat T cells increased in accordance with TCE exposure time,showing a time-effect relationship( P < 0. 01). CONCLUSION:s TCE at the level of 2. 00 mmol / L had no observed effect in Jurkat T cells. High doses of TCE( ≥5. 00 mmol / L) showed cytotoxic damages to naive and activated Jurkat T cells and low doses of TCE( ≤2. 00 mmol / L) could stimulate activated Jurkat T cells secrete IL-2 in a dosedependent and time-dependent manner.