Objective To study the cell-cycle specificity of tamoxifen-induced apoptosis in the ER(+) breast cancer cells in vitro and in vivo. Methods ER(+) MCF-7 cell line (in vitro) and primary cultured ER(+) breast cancer cells (in vivo) were treated with tamoxifen, and the cell cycle specificity of cell apoptosis and the apoptotic rate were dertermined by flow cytometry. Results Both MCF-7 and primary cultured breast cancer cells were induced to apoptosis with G 0/G 1 phase specificity by tamoxifen. And the apoptotic rate in MCF-7 was higher than that in primary cultured breast cancer cells. Conclusion Tamoxifen could induced G 0/G 1 phase specific apoptosis in the ER(+) breast cnacer cells, and tamoxifen-induced apoptotic rate was higher in vitro than in vivo.

Objective To detect estrogen receptor and DNA index in breast cancer by flow cytometry. Methods 32 cases of fresh specimens of breast cancer resected by operation were used in this study. Single cells suspension was prepared from those specimens, and estrogen receptor expression was analyzed by flow cytometry. At the same time, the status of ploid and S phase cell ratio were determined. Results Flow cytometry analysis could measure the expression level of estrogen receptor in the fresh breast cancer specimens, which was more sensitive and needed less volume of tumor tissue compared with immuohistochemical method. The information on the status of ploid and S phase cell ratio were simultaneously obtained. Conclusion The detection of estrogen receptor in breast cancers by flow cytometry was more helpful for evaluating prognosis and selecting treatment.

BACKGROUND: Glucose metabolism disturbance, one of clozapine's adverse effects, has received increasing attention from endocrinologists. Insulin resistance is believed to be associated with clozapine-induced glucose metabolism disturbance. Does it have direct effects on secretion function of islets?OBJECTIVE: To investigate the effects of different concentrations of clozapine on the secretion function of pancreatic islets in vitro.DESIGN: A completely randomized controlled design for the experimental study.SETTING: Department ofPsychiatry, People's Hospital of Wuhan University.MATERIALS: The experiment was conducted in the Laboratory Center of Stomatology Hospital of Wuhan University from September 2003 to January2004. Three healthy male Wistar rats of clean grade were used.METHODS: [1] Classical collagenase digestion method was used to isolate and purify the rat islets of Langerhans. [2] Hank's solution containing 2 g/L bovine serum albumin and 3.3 mmol/L glucose was added for preincubation for 30 minutes. The supernatant was removed. The wells were divided into five groups with 12 wells in each group. The control group was added with 1 g/L dimethyl sulphoxide (DMSO) and 3.3 mmol/L or 16.7 mmol/L glucose, 1 mL per well. The four experimental groups were added with 1 mL 0.2 μmol/L, 1 μmol/L, 5 μmol/L, or 10 μmol/L clozapine apart from the above DMSO and glucose. Six wells in each group were incubated for 1 hour, and another six wells were incubated for 4hours. The supernatants of the different groups were collected and stored at-20℃ in the refrigerator for later testing. The above procedures were repeated three times. The insulin released into the medium supernatant was examined by radioimmunoassay. [3] One-way ANOVA was used to compare the differences between the experimental groups and control group; LSD-t test was used for multiple comparison.MAIN OUTCOME MEASURES: The level of insulin secretion in the supernatants in culture solution containing 3.3 mmol/L or 16.7 mmol/L glucose which was incubated for 1 hour or 4 hours.RESULTS: [1] At 3.3 mmol/L glucose, no difference in insulin secretion was found between the four clozapine groups and control group after 1-hour incubation (P ＞ 0.05). After 4-hour incubation, the level of insulin in clozapine groups decreased significantly [(0.92±0.4), (1.02±0.3),(1.06±0.4), (0.74±0.2), (1.66±0.4) mU/IEQ, P ＜ 0.05]. There was no significant difference in the volume of insulin among the four clozapine groups with different concentrations (P ＞ 0.05). [2] At 16.7 mmol/L glucose, the level of insulin at the four concentrations of clozapine did not differ from that of control group either after 1 hour or 4 hours of incubation (P ＞ 0.05).CONCLUSION: Clozapine inhibits basal insulin secretion, but the effect is not correlated with its dosage.

BACKGROUND: Among atypical antipsychotic drugs (AAPDs), clozapine has the greatest effect on glucose metabolism, while risperidone, between clozapine, olanzapine and traditional antipsychotics, affects glucose metabolism less severely than clozapine.OBJECTIVE: To investigate the path through which the most commonly used AAPDs influence glucose metabolism by comparing the effects of risperidone, clozapine, and the metabolites of clozapine, desmethylclozapine (DCLO) and clozapine N-oxide (CNO), on insulin secretion in vitro.DESIGN: A completely randomized controlled design.SETTING: Center of Public Health of People's Hospital, Wuhan University.MATERIALS: The experiment was conducted in the Experiment Center of Stomatology Hospital, Wuhan University, from September 2003 to January 2004. Three healthy male Wistar rats of clean grade were used.bovine serum albumin and 3.3 mmoL/L glucose was added into each well (1 mL) for preincubation for 30 minutes. Then the supernatants were removed. Six wells were set as one group, and there were five groups altogether, namely, control group, risperidone group, clozapine group, DCLO group, and CNO group. All the groups were added with 1g/L dimethyl sulphoxide (DMSO), 3.3 mmol/L or 16.7 mmol/L glucose per hole (1 mL).Rrisperidone group, clozapine group, DCLO group, and CNO group were also added with 1 μmol/L risperidone, clozapine, DCLO, or CNO, respectively. For each group, three wells were incubated for another 1 hour, and the other three wells were incubated for 4 hours. Supernatants were collected and stored at -20 ℃ in the refrigerator. The experiment was repeated for three times. The level of insulin in the supernatants before and after study were expressed as the median (M) and quartile (P25, P75). Mann-Whitney test was used for data comparison.MAIN OUTCOME MEASURES: The level of insulin in the supernatants in the 5 groups.tion (GSIS) was significantly higher than that of basal insulin secretion, either 1 hour or 4 hours after incubation [1.91(1.68-2.62), 2.21(1.59-3.05) μU/IEQ;1.05(0.71-1.15), 1.65(1.16-1.84) μU/IEQ, P ＜ 0.05], which suggested that ferences in the level of basal insulin secretion and GSIS in risperidone group and CNO group were not significant either 1 hour or 4 hours after incubation. The level of basal insulin secretion of clozapine group after 4-hour incubation was lower than that of control group [1.65 (1.16-1.84),1.08 (0.88-1.20) μU/IEQ, P ＜ 0.05]. The level of GSIS in DCLO group was significantly lower either after 1-hour or 4-hour incubation [1.15(0.84-1.32), 1.91(1.68-2.62) μU/IEQ;1.08 (0.62 -1.33), 2.21(1.59-3.05) μU/IEQ, P ＜ 0.05,0.01].CONCLUSION: Clozapine affects basal insulin secretion, and its metabolite DCLO inhibits glucose-stimulated insulin secretion in vitro.Risperidone does not cause impairment in insulin secretion.

Objective To study the relationship between serum hepatitis B virus covalently closed circle DNA (HBV cccDNA) as well as liver function and liver tissue pathological changes in children with chronic hepatitis B.Methods One hundred and twenty-four HBV-DNA positive children with hepatitis B were enrolled.Among 124 patients,65 cases were HBV carriers,59 cases were chronic hepatitis (mild in 31 cases,moderate in 18 cases and severe in 10 cases).HBV cccDNA in serum and liver function were detected,46 of which underwent liver biopsy and liver tissue inflammation and fibrosis grading classification was made.Results In moderate and severe cases,positive rates of serum HBV cccDNA (77.8％,100％) were higher than those of the HBV carriers and mild cases (32.3％,54.8％) (x2 =25.429,P ＜ 0.01),indicating more severe illness in children,detection rate of serum HBV cccDNA was higher.ALT,AST,and TBIL were higher in serum HBV cccDNA positive group than those of negative group[(95.6 ± 18.2) U/L vs (52.5 ± 17.7) U/L,(88.8 ±20.3) U/L vs (48.4 ±21.4) U/L,(68.4 ±24.6) μmol/L vs (28.3 ± 23.9) μmol/L](t =15.572,10.750,17.067,P ＜ 0.01).Serum HBV cccDNA and liver inflammatory activity and fibrosis showed no significant correlationship.Conclusion Serum HBV cccDNA is a sensitive indicator of viral replication,the more severe the disease situation,the peripheral HBV cccDNA detection rate is higher.But it is not entirely consistent with liver inflammation and fibrosis,so it can not completely reflect the degree of liver damage.

OBJECTIVETo investigate the pathogenic and clinical presentation and laboratory tests of bullous rash in hand, foot and mouth disease (HFMD) in Xi'an from January 2013 to December 2014 by retrospective analysis.

METHODA total of 224 specimens were collected from clinically diagnosed HFMD cases who were characterized by widespread mucocutaneous bullous reactions in Xi'an Children's Hospital from January 2013 to December 2014, the identification and subtyping of the isolates were conducted with real-time fluorescent quantitative RT-PCR. A retrospective analysis was performed to analyze the clinical presentation, laboratory tests and late follow-up problems of the HFMD.

RESULTIn the clinically diagnosed HFMD cases who were characterized by widespread mucocutaneous bullous reactions, 207 were caused by coxsackievirus A6 (CA6), accounting for 92. 4% of all cases with bullous, 4 were caused by enterovirus 71 (EV71), accounting for 1.8%, 10 were caused by coxsackievirus A16 (CA16), accounting for 4. 5%; 4 cases were negative for these viruses. In the cases positive for intestinal virus-nucleic acid, 130 were male, 90 were female; male to female ratio was 1. 44: 1, 203 were <5 years old, accounting for 92. 3%. Leukocytosis was found in 75 cases (34. 1%); high-sensitivity C-reactive protein (hsCRP) increased in 200 cases (90. 9%); elevated myocardial enzyme CK-MB was found in 35 cases (15. 9%), alanine aminotransferase increased in 15 cases (6. 8%); 187 cases had fever (85. 0%). None of the cases had serious complications such as encephalitis or myocarditis. In the course of the critical phase bullous rash or large vesicle-like changes, obvious itching, and facial rash appeared. After the fluid in the bullae was absorbed or the bullae ruptured or became ulcerated, scar formation and large areas of exfoliation occurred, with no effusion on the newly formed epithelium in the base, without significant pigmentation on later follow-up. In the late follow up process, 52 cases in CA6-positive patients (25. 1%) developed onychomadesis within 2-4 weeks after onset, 1 to 8 nails, an average of 4. 3 fell off, new nails grew, the nail bed showed no structural abnormalities and hyperplasia after falling off, the surface was smooth, had no hypertrophy, left no sequelae.

CONCLUSIONThe pathogen in HFMD characterized by widespread bullous reactions was mainly the CA6, this kind of HFMD was mainly mild type, with significant itching, later the bullae may have scar formation and skin exfoliation, in some cases onychomadesis may occur.

Child ; Enterovirus A, Human ; Enterovirus Infections ; pathology ; Exanthema ; pathology ; Female ; Fever ; Hand, Foot and Mouth Disease ; pathology ; Humans ; Male ; Pruritus ; Retrospective Studies

Recently, the droplet microfluidic system attracts interests due to its high throughput and low cost to detect and screen. The picoliter micro-droplets from droplet microfluidics are uniform with respect to the size and shape, and could be used as monodispensed micro-reactors for encapsulation and detection of single cell or its metabolites. Therefore, it is indispensable to characterize micro-droplet and its application from droplet microfluidic system. We first constructed the custom-designed droplet microfluidic system for generating micro-droplets, and then used the micro-droplets to encapsulate important amino acids such as glutamic acid, phenylalanine, tryptophan or tyrosine to test the droplets' properties, including the stability, diffusivity and bio-compatibility for investigating its application for amino acid detection and sorting. The custom-designed droplet microfluidic system could generate the uniformed micro-droplets with a controllable size between 20 to 50 microm. The micro-droplets could be stable for more than 20 h without cross-contamination or fusion each other. The throughput of detection and sorting of the system is about 600 micro-droplets per minute. This study provides a high-throughput platform for the analysis and screening of amino acid-producing microorganisms.
Amino Acids ; isolation & purification ; Microfluidic Analytical Techniques ; Microfluidics ; instrumentation

Objective It's not clear why the severity of hand-foot-mouth disease (HFMD) caused by enterovirus 71 (EV71) was varied clinically.In this study,the relation between the virus load of EV71 and the severity of HFMD was analyzed and the foundation for HFMD control and treatment were laid.Methods Two hundred and fortyseven patients with HFMD caused by EV71 from Jan.2012 to Sep.2013 in Xi'an Children's Hospital were enrolled and clinical data were collected,including age,gender,clinical symptoms,signs,routine blood count,fasting glucose,C-reactive protein and enterovirus.The EV71 virus loads of 247 patients with HFMD were measured by real-time PCR assay.Meanwhile,the effect of EV71 RNA on the severity of HFMD was analyzed.Results In this study,121 mild cases and 126 severe cases with HFMD limb shaking,fasting glucose,lymphocyte and neutrophil percentage were statistically significant between the 2 groups (all P ＜ 0.05).The counts of EV71 RNA from all of the 247 throat swab samples were more than 100 copies/mL,and the highest virus load was 1.1 × 105 copies/mL.The highest value of EV71 RNA was 9.32 × 104 copies/mL and 1.1 × 105 copies/mL in mild and severe cases,respectively,there was no significant difference between the 2 groups (P ＞ 0.05).Conclusions The results of EV71 RNA by RT-PCR are visualized and convenient.The virus loads of EV71 are not associated with the severity of HFMD,but further studies need to analyze.

Background and purpose:Diffusion-weighted imaging (DWI) is a non-invasive technique of breast magnetic resonance imaging (MRI). DWI is an alternative to dynamic contrast-enhanced (DCE) MRI for differentiating malignant from benign lesions in breast screening or not. This study aimed to evaluate the potential role of DWI in differentiating malignant breast lesions from benign lesions.Methods:Seventy-four patients underwent digital mammography, DCE and DWI (49 patients’b-value of 0, 400, 600 and 800 s/mm2). The detectability, sensitivity and speciifcity of DWI and DCE were compared. Absolute apparent diffusion coefifcient (ADC) was compared with standardized ADC for quantitative analysis.Results:Sixty-four of 74 patients had positive pathologic findings (38 malignant, 26 benign). All of the malignant lesions were detected on DWI and DCE. The sensitivity of DWI was 83.33%, 90.00% and 93.33%, and the specificity was 85.91%, 76.19% and 72.72%, forb-value of 400, 600 and 800 s/mm2, respectively. The sensitivity and speciifcity of DCE were 86.61% and 90.48%. There was no signiifcant difference between absolute and standardized ADC in detecting breast cancer (P>0.05).Conclusion:DWI is an important complemented technique to DCE-MRI for differentiating malignant from benign lesions in breast MRI.