Objective:To detect karyotype homology of vaginal isolates from patients with recurrent vulvovaginal candidiasis (RVVC) in recurrent episodes, and to discuss changes of susceptibility of Candida strains to antifungal drugs with clinical progress.Method:s Ten patients were recruited from Beijing Obstetrics and Gynecology Hospital, Capital Medical University from September 2018 to June 2019, who were firstly diagnosed with RVVC. Vaginal discharges were collected before first treatment and after first relapse. Vaginal strains were isolated, purificated and identificated. Then karyotype of 20 strains isolated from 10 patients were detected by restriction endonuclease analysis of genomic DNA (REAG) using enzyme BssHⅡand pulsed field gel electrophoresis (PFGE) methods, and sensitivity of clinical isolates to 5 antifungal drugs (clostridium, fluconazole, miconazole, itraconazole and nystatin) was also detected using disk diffusion method. Result:s (1) All 20 strains of 10 patients with RVVC were Candida albicans, and their chromosomes were extremely similar after BssHⅡ enzyme digestion. The gene bands of isolated strains from the same patient were completely identical. (2) After clinical medication, the sensitivity of vaginal isolates to azoles was generally decreased, but remained highly sensitive to nystatin, nystatin (first and second clinical isolates: 100% sensitivity and 100% sensitivity)>clotrimazole (100% sensitivity and 90% sensitivity)>fluconazole (80% sensitivity and 70% sensitivity)>itraconazole (60% sensitivity and 50% sensitivity)>miconazole (30% sensitivity and 20% sensitivity). Conclusions:(1) The latency of the same colonized strain in the vagina may be the cause of repeated RVVC episodes. (2) Antifungal agents could selectively induce drug resistance to Candidas, and Candidas show cross-resistance to antifungal agents. Repeated fungal culture and drug sensitivity test in patients with RVVC are very necessary for correct selection of antifungals.

Objective To study the disease process of vulvovaginal candidiasis (VVC) infection in rat model of VVC, and to study the immuno-repairing effect of different treatments on vaginal epithelium and the ultra-structural changes of vaginal epithelial cells. Methods The VVC model of female rats were established. After successful modeling, the rats were treated with no treatment (model control group), nystatin and Kangfu Xiaoyan suppository. The vaginal epithelium was observed by transmission electron microscopy and immunohistochemical staining. The ultra-structural changes of epithelial cells and the expression of cytokines interferon γ (IFN-γ), interleukin (IL) 4, IL-17 and IgG in epithelial cells were observed and analyzed statistically. Results The negative conversion rate of model control group was 0, and that of nystatin group was 6/6, and that of Kangfu Xiaoyan suppository group was 5/6; significant difference existed between nystatin, Kangfu Xiaoyan suppository group and model control group (P<0.05). The ultrastructures of vaginal epithelial cells were damaged obviously after VVC infection, and the ultrastructures were repaired by nystatin and Kangfu Xiaoyan suppository under transmission electron microscope. Immunohistochemical staining showed, the expressions of IFN-γ and IgG in the four cytokines which played a protective role increased after Kangfu Xiaoyan suppository treatment, significantly different from that of model control group (P<0.05), but there were no significant differences of the IFN-γ and IgG expression between Kangfu Xiaoyan suppository group and those of nystatin group (P>0.05); the expression of IL-17 was increased after nystatin treatment, but decreased after Kangfu Xiaoyan suppository treatment, and the difference between the two groups had statistical significance (P<0.05). Conclusions The ultrastructure of vaginal epithelial cells after VVC infection could be damaged obviously, the local immune state is disordered, and the antifungal drug nystatin has a good therapeutic effect on VVC, it could significantly repair the damaged vaginal epithelium structure after VVC infection and strengthen the protective immune function of vaginal epithelium. Kangfu Xiaoyan suppository, one of Chinese medicine, has similar therapeutic effect with nystatin.

Objective To observe the influence of insulin resistance(IR) and isosorbide mononitrate(ISMN) on myocardial cellular apoptosis in spontaneously hypertensive rats (SHR) .Methods Forty male 14‐week old Wistar(W) rats and SHR(S) each were respectively or jointly fed with normal diet (ND) ,high fat and high glucose(HFHG) diet ,normal saline(NS)and ISMN by ga‐vage .Then they were randomly divided into the normal and NS group (normal W and normal S ) ,HFHG and NS group(HFHG W and HFHG S) ,normal and ISMN group(ISMN W and ISMN S) ,HFHG and ISMN group(HI W and HI S) ,with 10 rats in each group .After 12‐week feeding ,carotid arterial blood was collected for detecting blood glucose concentration and insulin level and cal‐culating insulin resistance index (HOMA‐IR);4 myocardial tissue samples were taken for respectively observing the morphology under microscope ,and detecting the NO level ,myocardial Bcl‐2 ,Bax gene and their protein expression levels in myocardial tissue . Results Myocardial NO level ,Bax gene mRNA and related protein levels in the HFHG and ISMN intervention groups were higher than those in the normal group ,while the bcl‐2 gene mRNA and related protein expression were on the contrary ;myocardial tissue NO level ,Bax gene mRNA and related protein expression in the S groups were increased compared with the corresponding W groups ,while the bcl‐2 gene mRNA and related protein expression were on the contrary ;in the HFHG W group ,the myocardial tis‐sue NO level had significantly positive correlation with HOMA‐IR ,and in the ISMN W group ,HOMA‐IR was positively correlated with the NO level in the myocardial tissue .Conclusion Myocardial cellular apoptosis of SHR is increased compared with Wistar rats ;both IR and ISMN can aggravate the apoptosis of SHR myocardial cells ,moreover IR has a mutual induction and reciprocal causation with ISMN .

OBJECTIVETo investigate the intramural lateral spread distance in low rectal cancer in order to provide basis for safety lateral resection margin of pull-through conformal resection (PTCR).

METHODSThe patients with low rectal cancer who received low anterior resection or abdominal-perineal resection in Changhai Hospital from December 2015 to March 2016 were enrolled and Surgical specimens were collected. After the specimens were fixed in 10% formaldehyde for 24 hours, a piece of tissue that was 1.5 cm in length and 0.5 cm in width from the edge of tumor was cut. The tissue was obtained in the direction of 3, 5, 7 and 9 o'clock clockwise. The distance of intramural lateral spread was measured in the specimens and the risk factors were analyzed.

RESULTSA total of 83 specimens were collected and the overall proportion of intramural lateral spread was 71.1%(59/83). The rate of lateral spread from 3 to 9 o'clock was 34.9%(29/83), 26.5%(22/83), 32.5%(27/83) and 37.3%(31/83) respectively, and the difference was not statistically significant(χ=2.444 9, P=0.485 3). The median distance of lateral spread in each direction was all 0 mm and the quartile range was 1 mm, 0.5 mm, 0.55 mm and 1 mm respectively. The 5th percentile (P5) of each direction was all 0 mm and the 95th percentile(P95) of each direction was 2.5 mm, 1.6 mm, 2.6 mm, 2.5 mm, respectively and the difference was not statistically significant either(χ=5.331 0, P=0.148 9). The rate of lateral spread of T1, T2, T3 and T4 was 0/4, 58.3%(14/24), 83.0%(44/53) and 1/2 respectively, and there was significant difference(P=0.005 0). The multivariate analysis indicated that T stage (P=0.002 2, OR=3.741, 95% CI: 1.606-8.716) was the risk factor of intramural lateral spread.

CONCLUSIONSThe intramural lateral spread does exist in low rectal cancer and T stage is the risk factor of lateral spread. The lateral resection margin should be 5 mm from the tumor edge at least when PTCR is performed.

Digestive System Surgical Procedures ; methods ; Humans ; Margins of Excision ; Multivariate Analysis ; Neoplasm Invasiveness ; pathology ; Neoplasm Staging ; adverse effects ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; surgery ; Risk Factors

Objective:To analyze the consistency of HER-2 expression among endoscopic biopsy and radical operation specimens of gastric adenocarcinoma and to investigate the clinical application value of HER-2 detection in trastuzumab treated patients. Methods:From March 2013 to February 2014, 167 patients from Shanghai Changhai Hospital were diagnosed with gastric adenocarcinoma using endoscopic biopsy specimens. The corresponding surgical specimens were collected for pathological analysis. The relevant clinical and pathological data were collected. HER-2 protein expression of endoscopic biopsy specimens was detected by immunohistochemistry (IHC). HER-2 protein expression and gene amplification status of the corresponding tumor resection specimens were detected by IHC and fluorescence in situ hybridization (FISH). Results were analyzed for clinicopathological characteristics. Results:Among the 167 cas-es, 18 cases (10.8%) were HER-2 positive, including 10 cases showing IHC3+and 8 cases showing IHC2+with FISH positive. The consis-tency rate result among endoscopic biopsy and surgical operation specimens was 82%. Excluding the cases showing IHC2+, the true positive rate and the true negative rate were 73.3%and 97.0%, respectively. Conclusion:HER-2 detection of endoscopic biopsy speci-men by IHC shows great predictive value. The main reason for the difference of surgery and biopsy specimens is the heterogeneity of tumor expression. Increasing the number of specimens and combined testing with FISH are important methods to reduce misjudge-ment.

BACKGROUND:Construction of tissue engineering adult cardiac myocytes has been a new research hotspot in cardiovascular fields.
OBJECTIVE:To explore a simple, fast method for the separation of adult rat cardiomyocytes, and preliminarily explore the construction methods of tissue engineering adult cardiac myocytes.
METHODS:Segmented enzymatic digestion method was used to isolate cardiac myocytes from adult rats. Subsequently, cardiac myocytes were transfected with adenovirus and liposome-mediated red fluorescent protein gene. Construction efficiency of tissue engineering cells was qualified using inverted fluorescence microscopy and flow cytometry. Final y, cardiac myocytes were transfected with adenovirus-mediated hypoxia-inducible factor-1a, and the expression of hypoxia-inducible factor-1a was examined by western blot analysis.
RESULTS AND CONCLUSION:A lot of cardiac myocytes were col ected using the segmented enzyme digestion method. Flow cytometric analysis showed that the survival of adult cardiomyocytes was (87.03±0.70)%. Compared with liposomal transfection (transfection efficiency was 0), adenovirus infection efficiency was (70.31± 1.39)%, and the cells expressed red fluorescence under fluorescence microscope. After 4 days of adenovirus infection, transfected cells expressed hypoxia-inducible factor-1a protein. These results showed that segmented enzyme digestion is a fast way to isolate adult cardiac myocytes, and recombinant adenovirus vector is a good vector to transfect cardiac myocytes.

Propionic acid, a major inhibitor to yeast cells, was accumulated during continuous ethanol fermentation from corn meal hydrolysate by the flocculating yeast under stillage backset conditions. Based on its inhibition mechanism in yeast cells, strategies were developed for alleviating this effect. Firstly, high temperature processes such as medium sterilization generated more propionic acid, which should be avoided. Propionic acid was reduced significantly during ethanol fermentation without medium sterilization, and concentrations of biomass and ethanol increased by 59.3% and 7.4%, respectively. Secondly, the running time of stillage backset should be controlled so that propionic acid accumulated would be lower than its half inhibition concentration IC50 (40 mmol/L). Finally, because low pH augmented propionic acid inhibition in yeast cells, a higher pH of 5.5 was validated to be suitable for ethanol fermentation under the stillage backset condition.
Biomass ; Ethanol ; metabolism ; Fermentation ; Flocculation ; Propionates ; chemistry ; Yeasts ; metabolism

Objective: To explore the sensitivity of Kit or PDGFRA mutants related to gastrointestinal stromal tumor (GIST) to Gleevec.Methods: The recombinant plasmids of KIT Del559-560, KIT Ins IPYD579, PDGFRA D842V and PDG-FRA L839P gene mutants were transiently transformed into the CHO cells by liposome methods.Western blot was used to detect the expression of the related protein and their phosphorylated forms after the cells were incubated with Gleevec for 90 min.At 72 hours after incubation with Gleevec, MTT was used to detect cell proliferation.Results: Western blot results showed that Gleevec at 0.1 μM can notably reduce phosphorylation of KIT Del559-560.Gleevec at 1μM completely blocked phosphorylation of KIT Ins IPYD579 and PDGFRA L839P, but did not affect PDGFRA D842V phosphorylation.MTT analy-sis indicated that growth of CHOPDGFRA L839P was inhibited by Gleevec at 1μM, however, CHOPDGFRA D842V was re-sistant to Gleevec at 5 μM.Conclusion: Gleevec can decrease the expression of phosphorylated protein CHOPDGFRA L839P and CHOKIT Ins IPYD579, and can remarkably inhibit the proliferation of cells containing PDGFRA L839P mutant.

Objective To design and implement a portable electrocardiogram (ECG) monitor for multiple users. Methods Multilevel menu on liquid crystal displayer(LCD) for switching among users based on digital signal processor(DSP) was developed. Record header in ECG records for different users was appended, and communication protocol between monitor and hospital center was made. Results This monitor could record three users’s ECG signal and transmit the ECG records to remote hospital center via digital communication method. The hospital center could receive and file the ECG records of different users. Conclusion This monitor can be used for three users and is more efficient in the application of ECG monitoring.