Objective:To summarize the efficacy and safety of transurethral partial cystectomy combined with intravesical suturation in the treatment of bladder cancer.Methods:The clinical data of 28 patients with bladder cancer who were admitted in Zhengzhou People's Hospital from July 2020 to October 2021 were analyzed retrospectively. There were 22 males and 6 females, with the average age of (68.8±8.9) years old. Twenty cases were with single bladder tumor, and 8 cases were with multiple bladder tumors. The diameter of single bladder tumor measured by CT was less than 4 cm, with a median tumor diameter of 2.3(0.9, 2.8)cm. All 28 patients underwent transurethral partial cystectomy combined with intravesical suturation for the first time. Intraoperative intravesical instillation was conducted with 1 g gemcitabine. Bladder hot reperfusion treatment was performed with gemcitabine 2 g at 45℃ immediately after operation. The patients underwent reexamination with cystoscopy every 3 months after operation.Results:All the 28 patients successfully completed the operation. The average operation time was (128.3±16.3)min. No obturator nerve reflection was induced during operation. Muscularis tissue was found in all tumor pathological specimens after operation, among which 4 cases showed muscularis infiltration. Pathology of tumor basal and peripheral tissues showed tumor cell infiltration in 3 cases (1 case full-thickness infiltration, namely the incision margin was positive). The median continuous irrigation time of bladder after operation was 24.0(20.9, 25.0)h. The median time of indwelling catheter after operation was 6.5(6.0, 7.0)days, and one case developed severe bladder spasm. All patients were followed up, with a median follow-up of 7.5(4.0, 10.8) months.Postoperative recurrence occurred in 3 cases, all of which were ectopic recurrence in bladder.The 1-year recurrence rate was 10.7% (3/28).Conclusions:In the first surgical treatment of bladder cancer, transurethral partial cystectomy combined with intravesical suturation can provide accurate pathological staging, and low positive surgical margin rate. Immediate suturation of bladder wound during operation and immediate bladder perfusion chemotherapy can provide short bladder irrigation time, with few adverse reactions and low incidence of complications.

ObjectiveTo determine the epidemiological characteristics of norovirus infections in Lin’an District， Hangzhou City from 2012 to 2020， and provide scientific evidence for improving preventive and control measures. MethodsDescriptive epidemiological methods were used to characterize the epidemic and conduct statistical analysis to determine related factors. ResultsA total of 37 clustered outbreaks of norovirus infection were reported in Lin’an District of Hangzhou from 2012 to 2020， including 8 outbreaks in kindergartens， 15 ones in elementary schools， 8 ones in middle and high schools， 2 ones in universities， and 4 other ones. The total number of cases with norovirus infection was 1 194， with the average attack rate of 3.76%. The incidence of norovirus was higher in winter and spring. It was also higher in urban areas， followed by suburban and mountainous areas neighbored to traffic lines. Moreover， attack rates differed significantly by transmission routes， including mixed contact and aerosol transmission， contact transmission， food-borne transmission， and water-borne transmission （χ²=186.91，P<0.001）. There was significant difference in the incidence among kindergartens， schools， and universities （χ²=980.15，P<0.001）. In the 37 outbreaks， norovirus were mainly classified as GⅡ in the 34 ones （accounting for 91.89%）， and GⅠ in the remaining 3 ones. ConclusionThe epidemic of norovirus infection in Lin’an District， Hangzhou City is characterized by certain population， time， space， transmission routes， and strains. It warrants enhanced health education and promotion， preparedness and response plan， syndromic surveillance， early alerting， school closure， and environmental disinfection for further prevention and control of norovirus infection.

Objective To study the effect of intestinal bacteria on motor ability of amyotrophic lateral sclerosis (ALS) mice models and its mechanism. Methods Twenty wild type C57BL/6J mice (WT group) and 20 SOD1-G93A transgenic ALS mice (ALS group) were selected as the research subjects. (1) Ten mice in both WT group and ALS group were selected, respectively; 5 mice in each group were fed in SPF environment, and the remaining 5 mice were fed in aseptic environment; they were defined as WT+SPF group, WT+aseptic group, ALS+SPF group and ALS+aseptic group. (2) Ten mice in WT group and ALS group were fed in sterile environment; 5 mice in each group were transplanted with fecal bacteria, and the remaining 5 mice were not interfered; they were defined as WT+transplantation group, WT+non-transplantation group, ALS+transplantation group and ALS+non transplantation group. The grip strength of mice was measured by grip force meter, the motor coordination ability of mice was tested by roller treadmill and rotating rod test, the number of motor neurons in the anterior horn of spinal cord was measured by Nissl staining, the expression of microglia activation marker ionic calcium junction protein (IBA-1) in spinal cord tissues was detected by immunohistochemical staining, and the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in spinal cord tissues were detected by Western blotting; the β-N-methylamino-L-alanine (BMAA) expression was detected by high performance liquid chromatography-tandem mass spectrometry. Results (1) The grip strength, drop latency and drop time of ALS+aseptic mice were significantly higher than those of ALS+SPF mice, the number of Nissl-stained positive cells was significantly larger than that of ALS+SPF mice, the number of IBA-1 positive cells was significantly smaller than that of ALS+SPF mice, the levels of TNF-α and IL-6 protein expressions and BMAA concentration were statistically lower than those of ALS+SPF mice (P<0.05). (2) The grip strength, drop latency and drop time of ALS+transplantion mice were significantly lower than those of ALS+non-transplantation mice, the number of Nissl-stained positive cells was significantly smaller than that of ALS+non-transplantation mice, the number of IBA-1 positive cells was significantly larger than that of ALS+non-transplantation mice, the TNF-α and IL-6 protein expressions and BMAA concentration were significantly higher than those of ALS+non-transplantation mice (P<0.05). Conclusion Imbalance of intestinal bacteria homeostasis can decrease the motor ability of ALS mice, which is related to the activation of microglia.

Objective: To study the mechanism of interleukin (IL)-17 in mice with non-alcoholic fatty liver disease for promoting M1-type macrophage polarization to exacerbate liver inflammation, and to provide references for the mechanism of NAFLD occurrence and development. Methods: A mouse model of NAFLD was constructed by high-fat diet. Mice were divided into control group, model group, IL-17 group, and anti IL-17 group. Histopathological changes of the liver were observed by HE staining. The serum levels of ALT and AST in peripheral blood of mice was detected by chemical colorimetry. Macrophages labeled with F4/80-PE, CD11C-FITC was designated as M1-type macrophages, those labeled with F4/80-PE, and CD206-APC was designated as M2-type macrophages. The proportion of M1 and M2 macrophages infiltrated into the liver tissues of mice were measured by flow cytometry. CD168 expression level of liver tissues was detected using immunohistochemistry. Protein and mRNA levels of the marker molecules (iNOS, TNF-alpha and IL-6) of M1 macrophages were detected using ELISA and RT-Q PCR. Western blot was used to detect the protein expression of JAK-STAT signal pathway and the expression level of MCP-1. Data were analyzed using one-way ANOVA and t-test. Results: High-fat diet NAFLD mice model was successfully constructed. IL-17 had increased the proportion of M1 macrophages in mice liver tissues and decreased the proportion of M2 macrophages (P < 0.05). The proportion of M1 and M2 macrophages in the liver tissues of normal mice was 7.9% ± 1.1% and 19.2% ± 1.8%. The proportion of M1 and M2 macrophages in the model group was 17.3% ± 2.5% and 15.0% ± 2.1. The proportion of M1 macrophages (33.8% ± 4.2%) in IL-17 group was higher than model group, while the proportion of M2 macrophages (7.8% + 1.0%) in IL-17 group was lower than model group. Protein and mRNA marker levels of M1 macrophage (iNOS, IL-12, TNFα and IL-6) in liver tissues were significantly higher than model group, control group, and anti-IL-17 group (P < 0.05). The expression levels of JAK1, STAT1, MCP-1, and CD168 in mice liver tissues of IL-17 group had increased (P < 0.05). The levels of aspartate and alanine aminotransferases in peripheral blood of mice in IL-17 group were significantly higher than other three groups (P < 0.05). Conclusion IL-17 can promote M1-type macrophage polarization, and exacerbates the liver inflammatory response to accelerate the progression of NAFLD in mice.

To screen the differentially-expressed microRNAs (miRNAs) in mouse models with renal ischemia-reperfusion injury (IRI),aiming to offer foundation for unraveling the molecular mechanism of the incidence and progression of IRI.Methods The mouse models with acute IRI were established by renal artery clamping.Fifteen mice were divided into the IRI group and sham surgery group (E group).The animals in the IRI group were subdivided into the A group (45 min ischemia followed by 24 h reperfusion),B group (25 min ischemia followed by 24 h reperfusion),C group (45 min ischemia followed by 4 h reperfusion) and D group (25 min ischemia followed by 4 h reperfusion) (n=3 for each group).The severity ofIRI was evaluated by histological changes and renal function.The differentially-expressed miRNAs in the IRI mouse models at different ischemia time (25 and 45 min) and reperfusion time (4 and 24 h) were screened by using cluster analysis of miRNAs microarray data.The differential expression of miR-695 and miR-145 was validated by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR).Results Both histological changes and renal function confirmed that the IRI mouse models were successfully established.Compared with the sham surgery group,71 differentially-expressed miRNAs were detected in the IRI group including 30 down-regulated miRNAs and 40 up-regulated miRNAs.The results of qRT-PCR demonstrated that if the standardized expression level of miRNAs in the E group was 1,the relative expression levels of miR-695 and miR-145 were 11.82 and 0.31 in the IRI group (both P＜0.05),which were consistent with the chip results.Conclusions After renal IRI,different changes occur in the gene expression profile of miRNAs.These differentially-expressed miRNAs act as molecular biomarkers for renal IRI with potential clinical and scientific research values.

Objective To investigate the incidence trends,clinical features and prognosis of ischemic colitis (IC) in China,and improve the level of diagnosis and treatment of IC.Methods China National Knowledge Infrastructure and VIP Chinese Science and Technology Periodical Database were searched.The ending date of search was May 15,2014.Results A total of 324 references were found,which were published from 1982 to 2013.There were 9202 cases reported,3973 cases of males and 5229 cases of females,with the male/female ratio of 1 ∶ 1.32 and mean age of (63.6±7.8)years.The amount of references and cases reported began to rise after 2002.Estimated ratio of cumulative incidence was higher in the north than in other areas of China (x2 =1097.95,P=0.000).The most common IC accompanying diseases were hypertension,heart diseases,diabetes,hyperlipidemia and constipation.There were statistically significant differences in the accompanying diseases between different regions and different times.Drugs,enteroscopy,surgery and low blood volume might be the precipitating factors.Patients commonly complained of abdominal pain,diarrhea/ desiring to defecate and hematochezia.Computed tomography was feasible in detecting lesions.Colonoscopy was the main method for diagnosis.The lesions were most common located in the left half colon including sigmoid colon,descending colon and splenic flexure,with typically in a segmental manner.Pan-colon involvement or rectum involvement rarely occurred.Type of transient lesion was the predominant subtype,which was generally managed non-operatively with good prognosis.Different from the type of transient lesion,pathological changes in gangrenous type were located in the right half colon including hepatic flexure of transverse colon,ascending colon and ileocecal junction.Gangrenous type required prompt surgical intervention but the mortality was much higher.Conclusions Incidence of CI has been increasing year by year.Patients who complain of abdominal pain,diarrhea/desiring to defecate and hematochezia should be considered as IC,particularly in the elderly women and patients with cardiovascular disease.Diseased regions are mainly located in sigmoid colon,descending colon and splenic flexure.Transient colitis is the predominant subtype,which have good prognosis.

Objective To investigate effect of antibiotic standardized management on prophylactic application of antimicrobial agents in type I andⅡ incision.Methods The hospital implemented the rule of Clinical Use of Antibiotics Management Approach and 2013 National Clinical Use of Antibiotics Special Rectification Program of Activities, 200 clinical surgery effective surgical cases before implementation and 250 clinical surgical effective surgical cases after implementation were selected.The thyroid surgery and hernia repair surgery were chosen as type I incision, and cesarean section were chosen as type Ⅱ incision.The basic data of surgery classification and antibacterials use were collected, then the antibiotics utilization ratio, the timing of administration of antibiotics, postoperative use of antibiotics time and combination therapy were compared between pre-implementation and post-implementation.Results The condition in accordance with applicable standards of antibiotics post-implementation: the antibiotics utilization ratio of thyroid surgery was 0 and post-operative stopping time was within one day.There was no combination therapy of three types surgery ( thyroid surgery, hernia repair surgery and cesarean section).The application of antimicrobial agents improved significantly but did not reach criteria:antibiotics utilization ratio of hernia repair surgery was 62.00%, which was higher than the criterion of 30% in antibiotic standardized management.Withdrawal time of antimicrobial agents post-operation in hernia repair surgery was (1.01 ±0.15)d which was above the standard of 1d and in cesarean section was (2.02 ±0.25)d which was above the standard of 2d.Conclusion The hospital strengthens management and standardizd regulation of antimicrobial drugs could facilitate standardization in type I and Ⅱ incision, and improve the quality of hospital medication administration of antibiotics.

Objective To increase the awareness of the incidence trend of ischemic colitis and improve the level of diagnosis and treatment.Methods From January 2004 to December 2013,the clinical data of 89 in patients with ischemic colitis were retrospectively analyzed.At the same period,100 patients diagnosed as chronic colitis were enrolled as control group.The laboratory findings of patients with ischemic colitis were compared with those of control group.Non normal distribution data were analysed by non parametric test.Results The annual person-times of patients with ischemic colitis and the percentages accounted in the whole hospital annual person-times increased year by year in recent 10 years,which was one person time and 0.01％ in 2004 increased to 41 person-times and 0.11％ in 2013.The average age of the 89 patients was (63.5±12.7) years,and the male to female ratio was 1∶2.18.The most common underlying disease in the 89 patients was hypertension,accounted for 43.8％.Thirty-eight patients had a history of abdominal surgery,and eighteen patients with appendectomy,accounted for 20.2％.Seventy-eight patients had abdominal pain and defecation desire.Seventy-four patients complained of hematochezia.Compared with control group,there was no statistically significant difference in fasting blood glucose,triglyceride,total cholesterol,blood platelet levels and prothrombin time (all P＞0.05).While the levels of C reactive protein and Ddimer were higher than those of control group (89.21 mg/L vs 61.29 mg/L,90.13 μg/L vs 59.39 μg/L; Z=-3.959,-4.377; both P＜0.05).The images of computed tomography (CT) indicated thickened bowel wall in lesion segment.The results of colonoscopy showed that in the left colon and rectum were the predilection sites,accounted for 93.88％ (138/147).The average hospital stay was (10.59 ± 5.97) days and t he recurrence rate was 3.37 ％ (3 / 89).The percentage of chronic cases was 2.25 ％ (2/89),and no death was reported.Conclusions The incidence of ischemic colitis significantly increased in recent years.Patients with abdominal pain and/or hematochezia should be considered,especially those over 60 years old.Receiving colonoscopy as earlier as possible was helpful for the diagnosis.

OBJECTIVETo establish a cell line with human NADH-cytochrome b5 reductase (b5R) deficiency via RNA interference (RNAi).

METHODSTwo siRNA expressing vectors targeting the b5R mRNA were designed and constructed. Hepatocellular carcinoma BEL-7402 cells were transiently transfected with the two recombinants by lipofectamine (TM) 2000, and semi-quantitative RT-PCR was carried out to analyze the suppression of b5R mRNA; BEL-7402 cells stably transfected with the two siRNA expressing vectors were selected in the media with G418. By analyses of the mRNA, enzymatic activity and protein level of b5R, several cell clones with deficiency of b5R were established. The cell growth curve of BEL-7402 cells with b5R deficiency was detected by MTT assay.

RESULTSTwo siRNA expressing vectors targeting b5R mRNA were obtained, namely pSib5R-1 and pSib5R-2. When BEL-7402 cells were transfected transiently with pSib5R-2, the expression of b5R mRNA was significantly suppressed with a suppression ratio of 68.3%, indicating that pSib5R-2 could trigger the degradation of b5R mRNA effectively. Eighteen clones stably integrated exogenous plasmids were obtained. In two clones from pSib5R-2 transfection, the expression of b5R mRNA was suppressed by up to 48.2% and 56.2%, and the enzymatic activity was inhibited by up to 54.6% and 63.5%, respectively. The protein levels also decreased significantly. The defect of b5R did not change the cell growth rate.

CONCLUSIONThe expression of b5R in BEL-7402 could be suppressed by vector-based RNA interference effectively. We established a cellular model with defect of b5R successfully, which can be used as a tool in investigation of the biological function of b5R and molecular mechanism of type II recessive congenital methemoglobinemia.

Cell Culture Techniques ; Cell Line, Tumor ; enzymology ; Cells ; enzymology ; Cytochrome-B(5) Reductase ; deficiency ; genetics ; Gene Expression ; drug effects ; Genetic Vectors ; Humans ; RNA Interference ; physiology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Transfection

OBJECTIVETo establish an in vitro heterogeneous expression model of human CYP2E1 (hCYP2E1) cDNA and investigate the effect of the chemical carcinogenic N, N'-dinitrosopiperazine (DNP) on the expression of CYP2E1.

METHODSExogenous hCYP2E1 was introduced into the mouse derived NIH3T3 cells using the lipofectamine transfection technique. Integration of exogenous hCYP2E1 gene was identified by PCR and Southern blot. After treatment with various concentration of ethanol and DNP on the transfected NIH3T3 cell cultures, RT-PCR and Western blot was applied to detect the expression level of CYP2E1.

RESULTSTwo cell clones with integration and stable expression of exogenous hCYP2E1 were obtained and designated as NIH3T3-2E1-A4 and NIH3T3-2E1-A8 respectively. The expression of both hCYP2E1 mRNA and protein products was promoted after either ethanol or DNP treatment.

CONCLUSIONThe results suggested that the promoted expression of hCYP2E1 induced by DNP and /or ethanol is due to enhanced transcription. The mechanism of DNP carcinogenes is might be related to this in situ activated metabolism by CYP2E1.

3T3 Cells ; Animals ; Carcinogens ; toxicity ; Cytochrome P-450 CYP2E1 ; genetics ; Ethanol ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Humans ; Mice ; Nitrosamines ; toxicity ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection