Objective To investigate the mechanism of action of Guizhi Fuling pill in treating chro-nic prostatitis(CP)using network pharmacology and molecular docking techniques.Methods Compo-nents of Guizhi Fuling pill were collected from the Traditional Chinese Medicines Systems Pharmacolo-gy Platform(TCMSP),and target information was obtained from the SwissTarget database.Targets for chronic prostatitis were screened from the GeneCards,OMIM,CTD,and DisGeNET disease data-bases.A protein-protein interaction(PPI)network was established and analyzed.Gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway en-richment analysis were performed using the DAVID database.The Cytoscape software was employed to construct an association network linking the components of Guizhi Fuling Pill,their targets,and the targets of chronic prostatitis.Molecular docking was conducted using AutoDock Vina software to verify the binding stability between the components of Guizhi Fuling pill and their targets.Results After screening and deduplication in the TCMSP database,76 components of Guizhi Fuling Pill were iden-tified,and 655 component targets were retrieved from the SwissTarget database.There were 190 intersecting targets between GuizhiFuling Pill and chronic prostatitis.GO analysis indicated that Guizhi Fuling Pill may treat chronic prostatitis by participating in processes such asapoptosis,ATP binding,and signal transduction.KEGG analysis suggested that Guizhi Fuling Pill can regulate pathways such as phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)and mitogen-acti-vated protein kinase(MAPK)to intervene in chronic prostatitis.Molecular docking data demonstra-ted that the components of Guizhi Fuling pill exhibited stable conformations with their targets.Con-clusion The components of Guizhi Fuling Pill can stably bind to their targets and exert therapeutic effects on chronic prostatitis through multiple targets and pathways.

Objective:To investigate the clinical effect of free deep femoral artery third perforating flap repaired soft tissue loss after Pilon fracture surgery in I stage.Methods:Fifteen patients were treated from April, 2013 to January, 2020. Miller AO classification: 8 cases 43-C1, 4 cases 43-C2 and 3 cases 43-C3. All cases were accompanied with severe soft tissue contusion and skin necrosis. After fracture reduction, soft tissue defects, internal fixation exposure and tendon exposure around the wound. Free deep femoral artery third perforating flap (3.5 cm ×15.5 cm to 5.5 cm×12.5 cm) for the repair of soft tissue defects around ankle in the I stage, the blood vessels of the flap were end-to-side anastomosed with vessels of the posterior tibial or anterior tibial. Regular follow-up after surgery.Results:One case of venous crisis occurred, other 14 cases survived, were followed-up from 5 to 18 months, the ankle joint function was good, did not affect the foot shoes, with excellent color and texture, the flap restored protective sensation, and leaving only linear scar, no muscle adhesion.Conclusion:Free deep femoral artery third perforating flap repaired soft tissue loss of surgical incision after fracture operated than significantly reduce the postoperative fracture infection and protect the blood supply around the fracture. It is an effective method of repair.

Objective:Evaluation of the clinical value of the BioPerfectus multiplex real time (BMRT)-HPV for cervical cancer screening.Methods:Physician-collected specimens of 1 495 women who were positive of Cobas 4800 HPV (Cobas-HPV), HPV genotyping based on SEQ uencing (SEQ-HPV), and (or) cytology ≥low grade squamous intraepithelial lesion (LSIL) in the primary screening of Chinese Multiple-center Screening Trial (CHIMUST), and 2 990 women selected from those who were negative of primary screening in the same project through nested control randomization with age-matching were tested for BMRT-HPV, which reported type-specific viral loads/10 000 cells in each specimen. With comparing to Cobas-HPV results and taking cervical histopathological diagnosis as the endpoint, the concordance of high-risk (HR)-HPV subtypes among the three assays was explored ,and the sensitivity and specificity of BMRT-HPV for cervical cancer screening were evaluated.Results:(1) The overall agreenment of HR-HPV subtypes between BMRT-HPV and Cobas-HPV, or SEQ-HPV test sample was 94.8%, 94.4%, with Kappa values 0.827, 0.814. (2) The sensitivity and specificity for cervical intraepithelial neoplasia (CIN) Ⅱ + of BMRT-HPV, Cobas-HPV and SEQ-HPV were 92.62%, 94.26%, 93.44% and 84.67%, 83.25%, 82.76%, respectively. There were no significant difference in sensitivity among the three HPV assays (all P>0.05), but the specificity of BMRT-HPV for CIN Ⅱ + was higher than those of Cobas-HPV and SEQ-HPV ( P<0.01). The sensitivity for CIN Ⅲ + of three HPV assays were all 100.00%, and the specificity for CIN Ⅲ + of BMRT-HPV was higher than those of Cobas-HPV and SEQ-HPV (83.40% vs 81.95%, 83.40% vs 81.50%; P<0.01). The number of pathological examinations of colposcopy for cervical biopsy detected in 1 case of CIN Ⅱ + or CIN Ⅲ + in BMRT-HPV was less than those in Cobas-HPV and SEQ-HPV ( P<0.01). When using HPV 16/18 + cytology ≥atypical squamous cell of undetermined signification (ASCUS) to triage HPV positive women among three assays, there was no different in the sensitivities of detecting CIN Ⅱ + and CIN Ⅲ + ( P>0.05). The specificity BMRT-HPV was slightly higher than those in Cobas-HPV or SEQ-HPV (all P<0.05), and the colposcopy referral rate was lower than those in Cobas-HPV and SEQ-HPV (all P<0.05). Conclusions:BMRT-HPV is as sensitive as Cobas-HPV or SEQ-HPV for primary cervical cancer screening, and has higher specificity. Therefore it could be used as a primary screening method for cervical cancer, which is worthy of clinical application.

Objective:To evaluate the value of p16 INK4a detected by p16 INK4a immunostaining as a new generation of cervical cytology for primary screening and secondary screening in population-based cervical cancer screening, and in improving cytological diagnosis. Methods:Between 2016 and 2018, 5 747 non-pregnant women aged 25-65 years with sexual history were recruited and underwent cervical cancer screening via high-risk (HR)-HPV/liquid-based cytological test (LCT) test in Shenzhen and surrounding areas. All slides were immuno-stained using p16 INK4a technology, among them, 902 cases were offered p16 INK4a detection during primary screening, and the remaining 4 845 cases were called-back by the virtue of abnormal HR-HPV and LCT results for p16 INK4a staining. Participants with complete LCT examination, HR-HPV test, p16 INK4a staining and histopathological examination results were included in this study. The performance of p16 INK4a in primary and secondary screening, and in assisting cytology to detect high grade squamous intraepithelial lesion [HSIL, including cervical intraepithelial neoplasia (CIN) Ⅱ or Ⅲ] or worse [HSIL (CIN Ⅱ) + or HSIL (CIN Ⅲ) +] were analyzed. Results:(1) One-thousand and ninety-seven cases with complete data of p16 INK4a and histology were included. Pathological diagnosis: 995 cases of normal cervix, 37 cases of low grade squamous intraepithelial lesion (LSIL), 64 cases of HSIL and one case of cervical cancer were found. Among them, 65 cases of HSIL (CIN Ⅱ) + and 34 cases of HSIL (CIN Ⅲ) + were detected. The positive rate of p16 INK4a in HSIL (CIN Ⅱ) + was higher than that in CINⅠ or normal pathology (89.2% vs 10.2%; P<0.01). (2) p16 INK4a as primary screening for HSIL (CIN Ⅱ) + or HSIL (CIN Ⅲ) + was equally sensitive to primary HR-HPV screening (89.2% vs 95.4%, 94.1% vs 94.1%; P>0.05), but more specific than HR-HPV screening (89.8% vs 82.5%, 87.7% vs 80.2%; P<0.05). p16 INK4a was equally sensitive and similarly specific to cytology (≥LSIL; P>0.05). (3) The specificity of LCT adjunctive p16 INK4a for detecting HSIL (CIN Ⅱ) + or HSIL (CIN Ⅲ) + were higher than that of LCT alone or adjunctive HR-HPV ( P<0.01), while the sensitivity were similar ( P>0.05). (4) p16 INK4a staining as secondary screening: p16 INK4a was significantly more specific (94.1% vs 89.7%, 91.9% vs 87.4%; P<0.01) and comparably sensitive (84.6% vs 90.8%, 88.2% vs 91.2%; P>0.05) to cytology for triaging primary HR-HPV screening. HPV 16/18 to colposcopy and triage other HR-HPV with p16 INK4a was equally sensitive (88.2% vs 94.1%; P=0.500) and more specific (88.3% vs 83.0%; P<0.01) than HPV 16/18 to colposcopy and triage other HR-HPV with LCT≥ atypical squamous cells of undetermined significance (ASCUS), and the referral rate decreased (14.0% vs 19.4%; P=0.005). Conclusions:For primary screening, p16 INK4a is equally specific to cytology and equally sensitive to HR-HPV screening. p16 INK4a alone could be an efficient triage after primary HR-HPV screening. In addition, p16 INK4a immunostaining could be used as an ancillary tool to cervical cytological diagnosis, and improves its accuracy in cervical cancer screening.

Objective:To explore the effectiveness and safety of holmium laser treatment via flexible bronchoscopy in children with tracheobronchial tuberculosis (TBTB).Methods:The clinical data of 40 children with TBTB admitted in Qilu Children′s Hospital of Shandong University from February 2016 to June 2019 were retrospectively analyzed. According to the treatment approach, they were divided into conventional treatment group (20 cases) and holmium laser treatment group (20 cases). The sex, age, course of disease and lesion location before treatment, and the time till relief of atelectasis and airway obstruction after treatment were compared between the two groups with t test and χ 2 test. Results:Among the 40 cases, 24 were males and 16 females, and 35 had lymph node fistula (87.5%) and 5 had granulation proliferation (12.5%). There was no statistically significamt difference between the conventional group and laser group in sex (male ratio: 50%（10/20） vs. 75%（15/20）, χ 2=2.66, P=0.10), age （(3.2±2.2） years vs. （2.2±1.8） years, t=1.41, P=0.16), course of disease (<1 month ratio: 30%（6/20） vs. 35%（7/20）, χ 2=0.11, P=0.73), lesion location (single lesion ratio： 45%（9/20） vs. 60%(12/20), χ 2=0.90, P=0.34). The effectiveness was evaluated after 3 months, and the total effective rate were 100% in laser group and 40% in conventional group, the difference was significant (χ2=25.34, P<0.01). No serious adverse events occurred in the two groups during the observational period. Conclusions:Holmium laser treatment via bronchoscopy is an effective way to remove the tuberculosis foci in the trachea, which can reduce the incidence of further related complications, and could be applied in clinical approach.

Objective To investigate the correlations of P2Y1 and ITGB3 single nucleotide polymorphisms (SNP) with aspirin resistance (AR) in patients with large atherosclerotic stroke (LAA) in a Chinese Han population.Methods Patients with first-ever LAA from Anhui stroke registration system were enrolled.Thrombus elasticity diagram was used to detect the platelet function.TaqMan technology was used to detect the P2Y1 and ITGB3 genotypes.Results A total of 206 patients with LAA were enrolled.Thirty-one patients (15.0％) had AR and 175 (85.0％) were aspirin sensitive (AS).The frequency of P2Y1 rs701265 G allele in the AR group was significantly higher than that in the AS group (43.5％ vs.26.9％;x2 =7.074,P=0.008).The frequency of P2Y1 rs701265 AA genotype in the AR group was significantly lower than that in the AS group (32.3％ vs.53.7％;x2 =4.850,P=0.028).There were no significant significances in the frequencies of P2Y1 rs1065776 and ITGB3 rs5918 alleles and genotypes between the AR group and the AS group.Multivariate logistic regression analysis showed that P2Y1 rs701265 G allele was an independent risk factor for AR in patients with LAA (odds ratio 2.186,95％ confidence interval 1.190-4.016;P=0.012).Conclusion The P2Y1 rs701265 polymorphism is associated with AR in Chinese Han patients with LAA,while the P2Y1 rs1065776 and ITGB3 rs5918 polymorphisms are not.

Objective: The aim of this work was to report the surveillance and dissemination of NDM-1 positive bacteria in a patient and ward environment. Methods: In 2010, during the therapy for a 51 years old patient, clinical and environmental samples were collected for carbapenem resistant bacterial culture, according to the clinical microbiological examination. Strains identification and antibiotic susceptibility were tested by VITEK Compact 2 system and E-test. The bla_NDM-1 was detected by PCR and analyzed by sequencing. Plasmids containing bla_NDM-1 were submitted to PFGE-S1 and Southern hybridization. Results: During hospitalization from October 1st to November 4th, nine strains were isolated from blood, sputum, urine, fecal, and ward ground samples. The Klebsiella oxytoca, Raoultella planticola, and Acinetobacter baumannii were isolated from blood sample. The Klebsiella pneumonia and Acinetobacter baumannii were isolated from sputum sample. An Acinetobacter lwoffii was isolated from urine sample. An Escherichia coli was isolated from fecal sample. And the Acinetobacter lwoffii and Acinetobacter spp. were isolated from ward ground. Four strains were NDM-1 positive, which were Raoultella planticola (RpNDM1) isolated from blood, Escherichia coli (EcNDM1) isolated from fecal, Acinetobacter lwoffii (AlDNM1) and Acinetobacter spp. (AsNDM1) isolated from ward ground. Four NDM-1 positive strains were resistant to Piperacillin, Piperacillin tazobactam, Cefepime, Ceftriaxone, Ceftazidime, Imipenem, Meropenem, and Ertapenem. Southern hybridization revealed that bla_NDM-1 were all located on plasmids in the four positive strains. Conclusion bla_NDM-1 can transfer rapidly among different species, resulting in difficult to control and prevent. While isolating patient who is carrying NDM-1 positive strains, more attention should be paid to the disposal of patient's excreta, especially stool, should be paid more attention.

Objective There is little research on the relationship of gene polymorphism of CYP2C19 and clopidogrel response after percutaneous transluminal angioplasty and stenting ( PTAS) in patients with ischemic cerebrovascular disease.The study aimed to investigate the relationship between gene polymorphism and high on-treatment platelet reactivity ( HTPR ) after PTAS and 6 months of regular dual antiplatelet administration in patients. Methods A total of 145 Chinese patients treated with PTAS in our de-partment from January 2011 to March 2014 were enrolled in this study.According to the gene sequencing, patients were divided into wild-type group(CYP2C19*1/*1,69 cases) and mutation group(heterozygous mutation CYP2C19*1/*2 and homozygous mutation CYP2C19*2/*2, 76 cases).Patients received a 100mg/d aspirin and 75mg/d clopidogrel maintenance dose (MD) as dual anti-platelet therapy after PTAS.The clopidogrel inhibition effect was measured by thrombelastography ( TEG) system 6 months after PTAS. Routine cerebral artery digital subtraction angiography was applied to evaluate whether there was restenosis in stent and logistic regres-sion analysis was used to analyze the influential factors of HTPR after PTAS and clopidogrel adminstration. Results After 6 months'regular administration of clopidogrel after PTAS, the platelet adenosine diphosphate ( ADP ) receptor inhibition rates in wild-type group, heterozygous mutation and homozygous mutation group were respectively (58.43 ±21.98)%, (47.80 ±22.93)%, (37.53 ± 21.84)%.The platelet ADP receptor inhibition rate was significantly decreased compared with wild-type group(P=0.001).Carriers of at least one CYP2C19 loss-of-function ( LOF) allele had a higher frequency of clopidogrel HTPR (35.5% vs17.4 % for patients with and without LOF alleles, respectively;P=0.014) .Using multivariate logistic regression analysis, the carriage of CYP2C19 LOF alleles was an independent predictor of the post-procedure HTPR (OR=2.356, 95% CI:1.053-5.272, P=0.037).The rate of ISR was remarkably higher in patients with at least one CYP2C19*2 alleles compared with wild-type patients(11.8%vs 1.4%, P=0.019) . Conclusion In patients with ischemic cerebrovascular disease, the CYP2C19 LOF allele had significant impact on post-procedure clopidogrel HTPR and the prognosis of ISR after PTAS.

OBJECTIVETo reveal the role of poly-ADP-ribosylation and DNA methylation in carcinogenic process induced induced by Cr (VI), and to discuss the relations between them.

METHODSThe pre-established Poly (ADP-ribose) glycohydrolase (PARG) deficient cells and 16HBE cells were treated with different concentrations of Cr (VI), and the changes of total genomic DNA methylation level in different groups were detected by methylation immunofluorescent detection, as well as the changes of the activity of methyltransferases. Moreover, RT-PCR and western blotting method were applied to analyze the changes of expression of DNMT1, DNMT3a, DNMT3b and MBD2, upon the protein level.

RESULTSAfter treated by Cr(VI) for 24 h, the healthy 16HBE cells showed a significant lower level of genomic DNA methylation; however, there was no significant changes (P > 0.05) found in PARG deficient cells by immunofluorescence assay. When the dose of Cr (VI) reached 5.0 µmol/L, the activity of methyltransferases in 16HBE cells and PARG deficient cells (49.33 ± 2.65, 80.05 ± 2.05) decreased by 20% and 50% comparing with contrast group (99.27 ± 1.10, 99.30 ± 0.60) . After treated by Cr (VI) for 24 h, the expression of mRNA and protein level among DNMT1, DNMT3a, DNMT3b and MBD2 decreased significantly in healthy 16HBE cells; and the expression of DNMT1 and DNMT3a decreased in PARG deficiency cells. The relevant expression levels of mRNA of DNMT1 were separately (0.99 ± 0.09), (0.79 ± 0.10), (0.59 ± 0.13) and (0.39 ± 0.02) (F = 247.17, P < 0.01), the expression levels of protein were separately (1.00 ± 0.03), (0.69 ± 0.15), (0.65 ± 0.10) and (0.55 ± 0.13) (F = 214.12, P < 0.01), the expression levels of DNMT3a mRNA were separately (1.00 ± 0.04) , (0.93 ± 0.11) , (0.79 ± 0.07) , (0.59 ± 0.05) (F = 498.16, P < 0.01) , and the expression levels of protein were separately (1.00 ± 0.14) , (0.97 ± 0.11) , (0.79 ± 0.17) , (0.57 ± 0.15) (F = 390.11, P < 0.01) when the dose of Cr (VI) at 0, 0.3, 1.2 and 5.0 µmol/L. However, there were no significant changes of expression found in DNMT3b and MBD2.

CONCLUSIONPoly-ADP-ribosylation could regulate the activity of DNMT3b and MBD2, protect cells against the DNA methylation alteration induced by Cr(VI) and maintain the global genomic DNA methylation level.

Cell Line ; Chromium ; toxicity ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; drug effects ; DNA-Binding Proteins ; metabolism ; Epithelial Cells ; metabolism ; Genome ; Humans ; Poly Adenosine Diphosphate Ribose ; metabolism ; RNA, Messenger ; genetics

Objective To investigate the effect of progressive neurological deterioration ( PND) of cerebral watershed infarction on early prognosis. Methods The consecutive patients with cerebral watershed infarction admitted in the Department of Neurology,Jinling Hospital,Nanjing University School of Medicine and their cerebral watershed infarctions confirmed by the imaging examination from March 2009 to March 2014 were enrolled. The clinical features, laboratory indicators and imaging features of internal watershed infarction,cortical-type watershed infarction,and mixed watershed infarction were identified and analyzed. The National Institutes of Health Stroke Scale was used to score neurological deficit. The modified Rankin scale ( mRS) was used to score the prognosis of patients. Single factor analysis was used to compare the differences between the groups. At the same time,the correlation between PND and poor prognosis of cerebral watershed infarction at day 90 was analyzed by multivariable Logistic regression analysis. Results A total of 89 patients with cerebral watershed infarction were enrolled,including 43 cortical-type watershed infarctions,36 internal watershed infarctions, and 10 mixed watershed infarctions. Single factor analysis indicated that the incidences of PND of internal watershed infarction and mixed watershed infarction were significantly higher than the cortical-type watershed infarction (36. 1% [n=13],50. 0% [n=5], and 16. 3% [n=7],respectively;P=0. 018). At day 90,28 patients had poor prognosis,and mRS was (3.4±1. 0) scores at day 90. There was significant difference in the types of infarction between the patients with poor prognosis and patients with good prognosis (P<0. 05). In patients with poor prognosis, most of them were internal watershed infarctions,accounting for 50. 0% (14/28),while in patients with good prognosis,most of them were cortical-type watershed infarctions(57. 4% [35/61]). The incidence of PND in patients with poor prognosis was significantly higher than that in patients with good prognosis (57.1% [16/28] vs. 14. 8% [9/61];P<0. 05). The result of multivariate Logistic regression analysis showed that after adjustment for confounding factor, PND was independently associated with the poor prognosis of cerebral watershed infarction at day 90 (OR 6. 969,95%CI 2. 451-19. 869;P<0. 01). Conclusion Compared with the cortical-type watershed infarction, the patients with internal watershed infarction is more prone to have PND, and PND is independently correlate with the poor prognosis at day 90.