Objective To develop a simple high-resolution melting ( HRM) analysis method for differentiation of Pc and P2 variants in class 1 integron.Methods DNA fragments containing Pc and P2 variants were amplified from plasmids pACW ( PcW ) and pACWP2 ( PcW-P2 ) respectively , then these purified PCR products and P 2 promoters were analyed full-length amplicon by HRM .Eight DNA fragments containing different Pc promoters were amplified and site-specific mutated from plasmids pACS ( PcS ) , pACH2 ( PcH2 ) , pACH1 ( PcH1 ) , pACW ( PcW ) , genomic DNA of Klebsiellar pneumonia HS07-68 (PcWTGN-10)and HS05-1792(PcH2TGN-10)respectively.The purified PCR products and eight Pc variants were characterized by HRM analyses of an unlabeled probe and full-length amplicon.This assay was applied to the differentiate Pc and P2 variants in 109 class 1 integrons from 95 urine clinical Escherichia coli isolates in Huashan Hospital during 2004 -2007.The differentiation results were compared with that determined by direct sequencing .Results P2 promoter with a significant higher melting temperature ( Tm ) can be identified by HRM analysis clearly .P2 promoters were identified in 2 class 1 integrons and consistent with direct sequencing results .Eight Pc variants were classified into three groups: PcS, PcSTGN-10 , PcW, PcWTGN-10, PcH1, PcH1TGN-10.Using direct HRM analysis.PcH2, PcH2TGN-10 were classified into four groups:PcS, PcH1, PcH2, PcW, PcSTGN-10 , PcH1TGN-10 , PcH2TGN-10 , PcWTGN-10 according to the melting curves of the unlabeled probe .Combined the HRM analyses of the whole amplicon and unlabeled probe , the eight Pc variants can be differentiated from each other .Five different Pc variants, PcS, PcW, PcH1, PcH2TGN-10 and PcWTGN-10 , were identified and consistent with direct sequencing results .Conclusions This developed a simple Pc and P 2 variants differentiation method via simultaneous HRM analyses of an unlabeled probe and full-length amplicon .This method is cost-effective and accurate , could be used in differentiation of Pc and P2 variants of class 1 integrons in clinical isolates .

Objective To investigate the changes in intracellular Ca2+ in the ureter smooth muscle cells (USMC) of rats with neuropathic urinary tract dysfunction (NUTD) and their significance.Methods Forty-five rats were randomly and averagely divided into NUTD group,experimental control (EC) group and blank control (BC) group.The NUTD group was operated with a spinal cord transection at the first lumbar level and the sacral cord was destroyed;in EC group the spinal process was partly bitten at the same position,but the spinal cord was not transected;BC group was given no operations.One week later,the video-urodynamic was performed to observe the acontractile detrusor (ACD),vesicoureteral reflux (VUR) and urinary tract dysfunction in rats among the NUTD group,EC group and the BC group.Video-urodynamic assessment was performed at the sixth week after operation.Ureter smooth muscle cells (USMC) were obtained by collagenase digestion.Intracellular Ca2 + in the USMC were observed by laser scanning confocal microscope.Then the effects of Bay K8644(10-8 mol/L,10-7 mol/L,10-6 mol/L) on cytosolic Ca2+ concentrations([Ca2+] i) in NUTD group were studied by calculating the fluorescence intensity.Results ACD and no detrusor overactivity were found in all rats in NUTD group and without vesicoureteral reflux.Immunofluorescence method confirmed that the cells were USMC.Compared with BC group (31.44 ± 2.82) and EC group (32.06 ± 3.67),the fluorescence intensity (FI) of intracellular Ca2 + in USMC was much lower in the NUTD group (9.80 ± 1.11),and there was significant difference(P ＜ 0.05).Bay K8644 (10-8 mol/L,10-7 mol/L,10-6 mol/L) increased the FI of [Ca2 +] i in a concentration-dependent manner,which were 3.80 ± 1.30,10.04 ± 2.15,19.89 ± 2.06,respectively,and there was significant difference (P ＜ 0.05).Conclusions The decrease in Ca2 + concentration in the ureter smooth muscle cells may be one of the important factors for the primary ureteral dysfunction of NUTD.And calcium channel agonist can be meaningful for adjusting abnormal Ca2+ concentration in USMC of NUTD.

The quality of SCI papers is one of the objective indexes of evaluation on scientific and technological strength and research capabilities.This paper introduced a comprehensive management strategy to promote the publication of SCI papers with high impact factors,in terms of such dimensions ass research orientation,financial and technical support,personnel training,and scientific research management platform.The short and long term effects of the comprehensive management strategy system were analyzed using the SCI papers publication data and IF data from 201 1 to 2014 at the hospital,as a reference for building a scientific management system of SCI papers for the administrators.

Objective:To explore the mechanism of action of Coptis chinensis in the treatment of dental caries using a network pharmacology approach and animal experiments. Methods:The active ingredients of C. chinensis and their targets were screened by the traditional Chinese medicine systems pharmacology (TCMSP) database and analysis platform, and the targets were searched online through the GeneCards database. The intersecting targets of C. chinensis and dental caries were screened at Venny 2.1, and the intersection targets were analyzed online for protein-protein interaction analysis and gene ontology (GO) and kyoto encyclopedia of genes and genomics (KEGG) enrichment. Then, Cytoscape was used to create a "component-target-pathway" network diagram. Rats were randomly divided into the model group and the C. chinensis group to establish a rat model of dental caries. Rats in the model group were repeatedly rubbed with a cotton ball soaked in 150 μl of 0.9% NaCl solution for 5 min, and rats in the C. chinensis group were repeatedly rubbed with a cotton ball soaked in C. chinensis (5.8 mg of C. chinensis in 150 μl of 0.9% NaCl solution) for 5 min. The two groups of rats were treated once a week for four consecutive weeks. The number of Streptococcus mutans colonies was counted, and serum serine/threonine protein kinase 1 (AKT1), JUN, interleukin-6 (IL-6), tumor necrosis factor (TNF), and B-cell lymphoma-2 (Bcl-2) were detected by enzyme immunoassay. Results:A total of 11 active ingredients in C. chinensis were found, which regulate multiple molecular pathways by intervening in 54 targets, thereby treating dental caries. Quercetin, berberine, flavodoxin, berberine infusion, and tetrahydroberberine were the core components, and AKT1, JUN, IL-6, TNF, and Bcl-2 were the core targets. GO analysis showed that BP mainly included cytokine activity, signaling receptor activator activity, signaling receptor modulator activity, cytokine receptor binding, and receptor ligand activity, etc.; and CC mainly included the response to lipopolysaccharides, the response to bacterial molecules, cellular responses to lipids, inflammatory responses, and negative regulation of cell population proliferation; MF mainly includes membrane rafts, membrane microregions, extracellular matrix, external encapsulated structures, and plasma membrane protein complexes, etc. KEGG analysis showed that advanced glycosylation end product-receptor for advanced glycosylation end products (AGE-RAGE), TNF, IL-17, Toll-like receptor, hypoxia-inducible factor-1 (HIF-1), mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB), epidermal growth factor receptor (EGFR), Janus kinase-signal transducer and activator of transcription (JAK-STAT), and phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathways have been associated with C. chinensis treatment. The results of animal experiments showed that serum Bcl-2 protein expression increased and serum AKT1, JUN, IL-6, TNF, and other proteins decreased after the C. chinensis treatment. Conclusions:C. chinensis can be involved in regulating the targets of dental caries through multiple pathways, with good therapeutic effects and a wide range of mechanisms of action, and is expected to be an important component in the development of proprietary Chinese medicines for the treatment of dental caries.

Objective To study the clinical value of whole －body magnetic resonance diffusion weighted imaging(WB－DWI) in evaluating the chemotherapy response for lung cancer,thus to provide evidence for optimizing clinical imaging examination. Methods From October 2017 to May 2018,60 patients with lung cancer confirmed by histopathology in Linfen Central Hospital were selected. The patients underwent DWI examinations before chemotherapy and after two cycles of chemotherapy. The change of tumor size,distant metastasis and apparent diffusion coefficient (ADC) value were compared before and after chemotherapy. The correlation between the change rate of ADC value and the shrinkage rate of tumor size in the effective group was analyzed. Results Of 60 cases,1 case had new cerebral metastases after chemotherapy. There were statistically significant differences in ADC value [(1. 12 ± 0.33) ×10 －3mm2/svs.(1.56±0.40) ×10 －3mm2/s]andtumorsize[(4.63±2.75)cmvs.(2.28±1.45)cm] between before and after chemotherapy in the effective group(t= －3. 954,4. 711,all P＜0. 01). There was correlation between the change of ADC value and tumor size(r=0. 34,P＜0. 05). Conclusion WB－DWI can not only detect the change of tumor size and distant metastasis quickly and effectively,but also can observe the microscopic changes of tumor cells by measuring ADC value. So it can predict the early therapeutic response of the tumor and make effective evaluation for the staging and chemotherapy response of lung cancer.

Objective To investigate the situation of sexual partners and related factors among men who have sex with men (MSM) in college students.Methods Snowball sampling and Convenience sampling were both used to recruit MSM from colleges in Tsingtao in 2016.Questionnaire-based interviews were conducted to collect data of socio-demographic and situation of sexual partners.Sample Size was estimated based on cross-sectional study,and theoretical 267.SPSS 17.0 software was used for statistical analysis.Results A total of 300 MSM,average aged 20.7,were analyzed.Both first sex partner and the last same-sex sexual partner were mct instantly,with proportions as 58.7％ (176/300) and 62.3％ (187/300) respectively.Among all the MSM,88.3％ (265/300) preferred selecting men as sex partners and 42.7％ (128/300) enjoyed finding sex partners in college,while 86.0％ (258/300) preferred finding their sex partners through intemet.Conclusions Intemet had been the major way of looking for sex partner among MSM in college students,the male sexual partner were met instantly.We should focus on men who have sex with men and their sexual health among college students to prevent and control HIV/AIDS.

Objective: To elucidate the clinicopathologic characteristics of atypical epithelioid trophoblastic lesions with cyst and fistula formation after cesarean section. Methods: The clinical and pathological data of 4 cases of post-cesarean atypical epithelioid trophoblastic lesions with cyst and fistula formation diagnosed at Women′s Hospital, School of Medicine, Zhejiang University during April 2007 to June 2018 were evaluated by hematoxylin and eosin stain and EnVision two-step immunohistochemical staining technique. Results: The age of the 4 patients ranged from 32 to 41 years, with a mean age of 36.5 years. Three patients recieved cystectomy and one underwent subtotal hysterectomy. Histologically, the lesions were well circumscribed and consisted of uniform cells of medium size, irregularly enlarged with hyperchromatic nuclei and 1 to 2 inconspicuous nucleoli embedded in abundant hyalinized matrix with fibrinoid material in the center. The cells exhibited immunohistochemical feature of chorionic-type intermediate trophoblastic cells (CK18+, p63+ and CD146-). All patients were alive without recurrence during follow-up of 1 to 40 months (mean=22 months). Conclusion Atypical epithelioid trophoblastic lesion with cyst and fistula formation after cesarean section has unique histological features, and its biological behavior and prognosis are still unclear, which need further exploration.

Objective:To explore the mediating role of interdisciplinary cooperation ability between the level of hospice care practice and difficulty degree of hospice care for medical staff, in order to improve the quality of hospice care services for medical staff, and to provide evidence for relevant managers to formulate effective measures to reduce the implementation of hospice care difficulties.Methods:This study was a cross-sectional survey. Using convenience sampling method, the general data questionnaire, Palliative Care Difficulties Scale (PCDS), Palliative Care Self-Report Practice Scale (PCPS) and Modified Index of Interdisciplinary Collaboration (MIIC) were used to investigate 362 medical staff in all hospice care institutions of Shiyan City from July to August 2022.Results:The total score of PCDS in 362 medical staff was (40.58±13.44) points, (67.47 ± 12.50) points for PCPS and (108.36 ± 21.46) points for MIIC. There was a positive correlation between the total score of MIIC and PCDS ( r=0.500, P<0.01). The total score of PCPS was negatively correlated with the total score of MIIC ( r=-0.337, P<0.01) and the total score of PCDS ( r=-0.189, P<0.01). The interdisciplinary cooperation ability of medical staff in hospice care had a complete mediating effect between the level of practice and the degree of difficulty (Effect value=-0.190, 95% CI-0.274 to -0.126), and the mediating effect accounted for 98.5% of the total effect value. Conclusions:The practice level of hospice care can not directly affect the implementation of the difficulty degree, but by taking active measures to improve the interdisciplinary cooperation ability, so as to enhance the practice level, and then reduce the implementation of hospice care difficulty degree.

Objective:To discuss the value of real-time three-dimensional speckle tracking imaging(RT3D-STI) in the evaluation of the left atrial (LA) volume and function in end-stage renal disease (ESRD) patients with different dialysis methods.Methods:Eighty-three ESRD patients who were treated in the Affiliated Hospital of Qingdao University from February to August 2019 were enrolled and divided into non-dialysis group ( n=25), hemodialysis group (HD group, n=31) and peritoneal dialysis group (PD group, n=27). And 30 healthy volunteers were enrolled as control group. Subsequently, the LA volume indexs including LA maximal volume index (LAVImax), LA minimal volume index (LAVImin) and LA presystolic volume index (LAVIpre), the LA function parameters including LA ejection fraction (LAEF), LA passive ejection fraction(LApEF) and LA active ejection fraction (LAaEF) and the LA global strain parameters including global longitudinal strain(GLS), global radial strain(GRS) and global circumferential strain(GCS) were obtained by RT3D-STI. The differences of the parameters among these groups were compared and the correlations between the myocardial strain parameters and dialysis time were analyzed. Results:①The LAVImax, LAVImin and LAVIpre of the non-dialysis group, PD group and HD group were higher than those in the control group, while the LAEF, LApEF, GLS, GRS and GCS were lower than those in the control group(all P<0.05); the LAVImin of the PD group as well as the LAVImin, LAVIpre and LAVImax of HD group were higher than those of non-dialysis group, while the LAEF, LApEF, GLS, GCS of the PD group and the LAEF, LApEF, GLS, GRS, GCS of HD group were lower than those of non-dialysis group (all P<0.05). The LAVImax, LAVImin, LAVIpre of HD group were higher than those of PD group, while the LAEF, LApEF, GLS, GRS and GCS of HD group were lower than those of PD group (all P<0.05). ②The GLS and GCS in PD group were negatively correlated with the dialysis time( r=-0.670, -0.596, all P<0.05), and the GLS, GCS and GRS in HD group were negatively correlated with the dialysis time( r=-0.624, -0.631, -0.551, all P<0.05). Conclusions:RT3D-STI can assess the LA volume and function of ESRD patients with different dialysis methods effectively and can further evaluate the prognosis of patients and guide timely clinical intervention.

Objective To understand the gender characteristics,HIV/AIDS related knowledge awareness and behaviors of transgender women.Methods A questionnaire survey was conducted among the transgender women recruited through snowball sampling in Jinan in 2014,and descriptive epidemiologic analysis was conducted on the survey results.Results A total of 55 transgender women were surveyed,all of them were male physically and female psychologically.Serious gender conflict occurred in 27 subjects (49.1％),and very serious gender conflict occurred in 8 subjects (14.5％).Thirty subjects dressed up as a man in social life,accounting for 54.5％;25 subjects dressed up as a women in social life,accounting for 45.5％.The average awareness rate of HIV/AIDS related knowledge was 57.9％(22/38).The awareness of knowledge about AIDS associated behaviors,such as multi sex partner and anal sex,was poor.For the lovers or sexual partners,58.2％ of the subjects (32/55) would choose males and 50.9％ of the subjects (28/55) had chosen males,and for the sex partner at latest sex,63.6％(35/55) of the subjects had chosen males.Up to 56.3％ of the subjects had sex with casual sexual partners (net friends and partners of one-night stand) at latest sex behavior.Among the subjects surveyed,18(32.7％) never used condoms;29(52.7％) used condoms occasionally;4(7.3％) used condoms frequently and 4(7.3％) used condoms at each sex.Conclusions AIDS associated high risk behaviors were common among the transgender women,such as unprotected anal sex,multiple sexual partners,frequent sex and poor condom use.It is necessary to conduct the study of the HIV infection prevention in transgender women.