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Objective:To investigate outcomes and safety of doxycycline-moxifloxacin sequential regimen in the treatment of Mycoplasma genitalium urethritis/cervicitis. Methods:From June 2019 to December 2020, patients with Mycoplasma genitalium urethritis/cervicitis confirmed by nucleic acid amplification testing were successively recruited at Department of Sexually Transmitted Diseases, Hospital of Dermatology, Chinese Academy of Medical Sciences, and received sequential therapy with oral doxycycline for 7 days followed by oral moxifloxacin for 7 days. Clinical and/or etiological assessment was conducted 2 to 3 weeks after the end of treatment. Fisher′s exact test was used to analyze factors influencing the treatment outcome. Results:Totally, 36 eligible subjects were enrolled, including 30 males and 6 females. Among them, 18 (50%) patients completed post-treatment etiological assessment, which showed that 12 achieved microbiological cure, and treatment failures occurred in 6; another 18 patients achieved clinical cure. The overall response rate to doxycycline-moxifloacin sequential therapy was 83.3% (30/36, 95% confidence interval[ CI]: 70.5%, 96.1%) . The treatment outcome showed no significant association with the patients′ age, gender, marital status, number of sexual partners in the past 1 month, history of sexually transmitted diseases, history of antibiotic use in the past 1 month, or co-infections (all P > 0.05) . Conclusion:The efficacy of doxycycline-moxifloacin sequential regimen is limited in the treatment of Mycoplasma genitalium infections in Nanjing area, and clinicians should be alerted to the possibility of treatment failure in clinical practice.
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Mycoplasma genitalium can cause non-gonococcal urethritis, cervicitis, and even epididymitis, pelvic inflammatory diseases, preterm birth and other complications. Due to increased prevalence of Mycoplasma genitalium infections and emergence of resistance to a variety of commonly used antibiotics in recent years, European experts, Australian Sexual Health Alliance (ASHA) and British Association for Sexual Health and HIV (BASHH) issued guidelines for the diagnosis and treatment of Mycoplasma genitalium infections in 2016, 2017 and 2018 respectively. These guidelines provide opinions on clinical manifestations and diagnosis of Mycoplasma genitalium infections, treatment and follow-up of patients and partners, which have an important role in guiding the management of Mycoplasma genitalium infections in China.
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AIM: To investigate the effect of curcumin on oxidized low-density lipoprotein (ox-LDL)-induced injury of human aortic endothelial cells (HAECs).METHODS: HAECs were pre-treated with curcumin at different concentrations and then treated with ox-LDL.The cell viability was assessed by MTT assay.The cell proliferation ability was analyzed by EdU assay.ELISA was used to determine the concentrations of interleukin-6 (IL-6), transforming growth factor β1 (TGFβ1), high mobility group box-1 protein (HMGB1) and secretory receptor for advanced glycation end products (sRAGE) in the HAEC culture medium.The binding activity of peroxisome proliferator-activated receptor γ (PPARγ) was evaluated by electrophoretic mobility shift assay.The protein levels of HO-1, HMGB1, RAGE,IL-6,TGFβ1 and phosphorylated PPARγ in the HAECs were determined by Western blot.RESULTS: The viability and the proliferation ability decreased significantly in the HAECs treated with ox-LDL.The PPARγ/HO-1 signaling pathway was inhibited while its down-stream HMGB1/RAGE signaling pathway was activated by ox-LDL.The levels of IL-6, TGFβ1, HMGB1 and sRAGE were increased.Pre-treatment with curcumin activated PPARγ/HO-1 signaling pathway and inhibited HMGB1/RAGE signaling pathway in ox-LDL treated HAECs in a concentration-dependent manner.The levels of IL-6, TGFβ1, HMGB1 and sRAGE were also decreased dramatically by pre-treatment of curcumin in a concentration-dependent manner.CONCLUSION: ox-LDL induces HAEC damage by inhibiting PPARγ/HO-1 to activate HMGB1/RAGE inflammatory signaling.Curcumin exerts protective effect on ox-LDL treated HAECs via activating PPARγ/HO-1 signaling pathway.
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Objective To investigate the serovar distribution of Chlamydia trachomatis (Ct) isolated from male patients with urethritis in sexually transmitted disease (STD) clinic.Methods Urine specimens were collected from male patients with urethritis in STD clinic at Hospital of Dermatology,Chinese Academy of Medical Sciences between January 2013 and December 2013.Fluorescence-based quantitative PCR was performed to detect Ct DNA in these specimens.DNA was extracted from Ct-positive urine specimens,and nested PCR was conducted to amplify the VS1-VS2 regions of the outer membrane protein A (ompA) gene,followed by gene sequencing.The resulting sequences were aligned to reference sequences by the DNAStar5.0 software to determine Ct serovars.Results A total of 432 urine specimens were collected,and 33.1％ (143/432) of them were positive for Ct.The VS1-VS2 regions of the ompA gene were amplified from 127 out of the 143 Ct-positive specimens,but not from the other 16 specimens.Nine serovars were identified by gene sequencing among the 127 specimens,including serovar E (29 strains,22.83％),F (28 strains,22.05％),D (19 strains,14.96％),G (16 strains,12.60％),J (16 strains,12.60％),K (8 strains,6.30％),H (5 strains,3.94％),I (3 strains,2.36％) and B (3 strains,2.36％),and Ct serovars E,F,D,J and G accounted for 85.02％ among all the strains.Synonymous mutations were identified in 14 out of the 127 strains when compared with reference strains.Conclusions E,F,D and G serovars were the main Ct serovars in male patients with urethritis in STD clinic.The proportion of Ct serovar E strain was decreased,but that of serovar J strain was increased compared with 20 years ago.
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Gonorrhea is a sexually transmitted disease caused by Neisseria gonorrhoeae,and substantially harms human health and socioeconomic development.Due to inappropriate treatment and the presence of drug resistance genes in patients,antibiotic resistance has emerged in Neisseria gonorrhoeae,such as resistance to penicillin,tetracycline,ciprofloxacin,or other antibiotics.Currently,extended-spectrum cephalosporins (ESCs) are the first-line treatment of gonococcal infection.With the wide use of ESCs,the sensitivity of Neisseria gonorrhoeae to ESCs has been decreasing gradually,and there have been reports on cases of treatment failure in clinical practice.In order to control gonorrhea and deal with drug resistance in Neisseria gonorrhoeae,combined therapy,alternative therapy and new drugs have been developed in clinic.
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Objective:Carcinoma of bladder is the most common malignancy in the urinary system in China.Most patients with this disease had non-muscle invasive bladder cancer (NMIBC)at the time of diagnosis.Radical cystectomy was indicated for patients with high risk or refractory NMIBC.We aimed to investigate the overall survival and disease-specific survival and related influence factors in patients un-dergoing radical cystectomy for pathological non-muscle invasive bladder cancer.Methods:From Jan. 2006 to Dec.2012,a total of 164 patients with pathological non-muscle invasive bladder cancer under-went radical cystectomy in Peking University First Hospital.Clinical data were retrospectively collected. Incidence of lymph node metastasis and disease recurrence were calculated.The risk factors of disease re-currence were analyzed.Kaplan-Meier plots were used to estimate the overall survival and cancer-specific survival.Multivariate Cox regression analysis was used to evaluate the prognostic factors for survival.Re-sults:Of all the patients included,159 had T1 disease,and 5 had CIS only.The median follow-up dura-tion was 46.5 months (range:7 -99 months).Fourteen patients were lost during the follow-up.Lymph node metastasis was noted in 6 patients (3.7%),4 patients had N1 disease,one patient had N2 di-sease,and one patient had N3 disease.Disease recurrence occurred in 16 patients (9.8%).The most common recurrence sites were the liver,bones,and lungs.The 5-year overall survival and disease-spe-cific survival for all the patients were 85% and 91%,respectively.The patients who underwent pelvic lymph node dissection showed a better prognosis in terms of disease-specific survival than those without (P =0.012).Patients with recurrence harbored a significant poorer survival (P <0.001).According to univariate Cox regression analysis,whether lymph node dissection was performed was an independent risk factor of disease recurrence (P =0.050,OR =2.695,95%CI 0.999 -7.271).In COX regression mo-del,age (P =0.008,OR =1.071,95%CI 1.018 -1.126)and whether lymph node dissection was performed (P =0.011,OR =3.385,95%CI 1.329 -8.621)were related to disease-specific survival. Conclusion:Patients with pathological non-muscle invasive bladder cancer underwent early radical cys-tectomy have a favorable prognosis,and bilateral pelvic lymph node dissection is essential for this proce-dure as it gains a survival benefit for the patients.
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Objective To establish two nested PCR assays for detection of Trichomonas vaginalis in urine samples from male patients with urethritis,and to evaluate their diagnostic value.Methods One thousand and eighty-eight male patients with urethritis were enrolled from sexually transmitted disease (STD) clinic in the Hospital of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College between April 2011 and December 2013.Urethral swabs were collected followed by smear testing,wet mount microscopic examination of Trichomonas vaginalis,and cultivation of Neisseria gonorrhoeae.Urine specimens were also obtained from these patients followed by DNA extraction.Two nested PCR assays were developed and performed to amplify the repeat genomic sequence and β-tubulin gene of Trichomonas vaginalis.Results Trichomonas vaginalis was detected in none of these swab specimens by wet mount microscopy,but in 29 (2.67％) of the urine specimens by either of the two nested PCR assays.Moreover,the positive specimens detected by the two nested PCR assays were completely consistent.Conclusion Compared with wet mount microscopy,nested PCR has higher sensitivity and specificity in detection of Trichomonas vaginalis in urine samples from male patients.
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Objective To evaluate the relationship between spectinomycin resistance in Neisseria gonorrhoeae and mutations in the rpsE gene.Methods Genomic DNA was extracted from 4 clinical isolates of Neisseria gonorrhoeae with different levels of spectinomycin resistance.Then,PCR was performed to amplify the entire rpsE gene and the spectinomycin resistance-determining region (SRDR) in the 16S rRNA gene followed by direct sequencing.Two spectinomycin-sensitive Neisseria gonorrhoeae strains were transformed with the genomic DNA containing the mutant rpsE gene.Subsequently,the susceptibility of the transformants to spectinomycin was determined,and PCR was performed to amplify the rpsE and 16S rRNA genes in the transformants followed by sequencing.Results All the 4 spectinomycin-resistant Neisseria gonorrhoeae strains harbored an A70C transversion in the rpsE gene,but no abnormality in the SRDR of the 16S rRNA gene.No mutations were detected in the spectinomycin-sensitive Neisseria gonorrhoeae strains.The A70C transversion in the rpsE gene was also detected in the two Neisseria gonorrhoeae transformants with spectinomycin resistance.Conclusion The A70C point mutation within the rpsE gene is associated with spectinomycin resistance in Neisseria gonorrhoeae.
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Objective To study the association of U. urealyticum biovar and genotype with nongonococcal, nonchlamydial mucopurulent cervicitis. Methods The study population consisted of two groups: patient group (226 female patients with nongonococcal, nonchlamydial mucopurulent cervicitis) and control group (118 healthy women). The biovar and genotype of U. urealyticum were identified in specimens positive for U. urealyticum culture by using PCR-single-strand conformation polymorphism (PCR-SSCP) analysis. Results The most common genotype in both groups was mba 3/14 in biovar 1 with the detection rate being 30.98%(57/184) in patients with mucopurulent cervicitis and 43.42% (33/76) in the controls. A significant difference was observed in the prevalence of genotype 2B in biovar 2 between the patients and controls [16.30% (30/184) vs 6.58% (5/76), χ2 = 4.367, P= 0.037). The genotype 1, 3/14 and 6 in biovar 1 predominated in the controls with their total prevalence being 81.58%. Conclusion The genotype 2B in biovar 2 of U. urealyticum may be associated with nongonococcal, nonchlamydial mucopurulent cervicitis among female patients attending an STD clinic.
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Objective To study the relationship between M. Genitalium and nongonococcal, non-chlamydial mucopurulent cervicitis. Methods A total of 226 patients with nongonococcal, nonchlamydial mu copurulent cervicitis and 118 healthy female volunteers were recruited in this study. Cervical samples were collected and M.genitalium was detected by PCR amplification of 16s rRNA and Pa genes. Information about socio-demographic characteristics, medical history, and sexual behaviors was gathered by questionnaire from both populations. Results The prevalence of M. Genitalium infection was 11.06% (25/226) in patients with mucpurulent cervicitis, 0,85% (1/118) in the healthy controls; the difference was significant between the two groups (x2 = 11.58, P < 0.001). Single variant analysis on the 226 patients showed that the preva-lence of M. Genitalium infection was 27.78%, 16.36%, 18.28% and 14.12% in patients with ectopic pregnancy history, cervical inflammation, pelvic organ tenderness, 10 or more polymorphonuclear leukocytes (PMNs)per oil immersion field in cervical discharge, respectively, significantly higher than that in patients without ectopic pregnancy history, cervical inflammation or pelvic organ tenderness, and those with less than 10 PMNs per oil immersion field in cervical discharge (9.62%, 6.03%, 6.02% and 1.79%, all P < 0.05). M.genitalium infection was also related to multiple sex partners and the presence of mucopurulent secretion in cervix (P < 0.001). Conclusion The prevalence of M.genitalium infection is higher in patients with non-gonococcal, nonchlamydial mucopurulent cervicitis attending STD clinic than that in normal population.
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Objective To investigate the amino acid patterns in penicillin-binding protein 2(PBP2)in Neisseria gonorrhoeae isolates with reduced susceptibility to ceftriaxonc.and the relationship between the amino acid patterns and reduced ceftriaxone susceptibility.Methods DNA was extracted from 13 clinical isolates of N.gonorrhoeae.including 11 strains with decreased susceptibility to ceftriaxone and 2 sensitive isolates.The full-length penA gene encoding the penicillin-binding protein 2 was amplified and sequenced.BLASTn and BLASTx programs were used to assess the insertion and substitution patterns of nucleotides in penA gene and of amino acids in PBP2,respectively.Results BLASTn analysis revealed insertion or substitution of 18-38 nucleotides in the penA gene of gonococcal isolates with reduced ceftriaxone susceptibility.As shown by BLASTX analysis.there were five patterns of amino acid substitution or insertion in PBP2 of the 11 isolates with reduced ceftriaxone susceptibility.However.mosaic structure of PBP2 was not found in any of these isolates.Conclusion Mosaic PBP2 seems not to be the major factor contributing to the decrease in susceptibility of N.gonorrhoeae to ceftriaxone.
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To study the cleavage of neuron stem cells in time lapse image sequences and realize their features abstraction, identification and tracking, a precise segmentation algorithm that can preserve the shape of division cells is presented in this paper. The fuzzy threshold segmentation is based on Zadth's maximum entropy. The optimal parameters of the maximum fuzzy entropy are decided by genetic algorithm. Region merging and splitting of the under-segmentation objects of the result of fuzzy segmentation are realized by weighted distance transform, region labeling and some operations on morphology. By comparison with some results of fuzzy and hard segmentation, this algorithm can implement the precise segmentation that is necessary for some specified objects in automatic identification and tracking of neuron stem cells.
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Humans , Algorithms , Cell Culture Techniques , Methods , Cell Division , Computer Simulation , Entropy , Fuzzy Logic , Image Processing, Computer-Assisted , Microscopy, Video , Methods , Models, Biological , Neurons , Cell Biology , Stem Cells , Cell BiologyABSTRACT
Objective To evaluate the ability of restriction endonuclease analysis by pulsed field gel electrophoresis (PFGE REA) and random amplified polymorphic DNA (RAPD) analysis for molecular typing of Neisseria gonorrhoeae isolates. Methods Genomic DNA from 36 Neisseria gonorrhoeae isolates were digested with SpeI and analysed by contour clamped homogeneous electric fields electrophoresis. The 36 isolates were also characterised by RAPD analysis with arbitrary primer OPA 03. Results 12～19 DNA fragments from 5 to 600kb were obtained among the 36 isolates after PFGE REA. The 36 isolates were discriminated into 20 PFGE patterns. Amplification of Neisseria gonorrhoeae DNA with primer OPA 03 produced 15 different DNA fragments (280～1900bp), and 3～9 fragments per strain could be seen. The 36 isolates showed 18 different RAPD patterns. Strains sharing common auxotypes and antibiotic spectrum could be differentiated by PFGE REA and RAPD analysis. General agreement was found between these two techniques. Isolates from different geographic areas and even some isolates in the same area showed considerable amount of DNA polymorphism. In addition, some isolates shared common or very similar patterns were unique to a given geographic area. Conclusion It is concluded that both PFGE REA and RAPD analysis are useful, sensitive molecular techniques for differentiation of clinical isolates of Neisseria gonorrhoeae. They should be helpful in the investigation of strain origin, clonal relation among strains and spread of antibiotic resistance. Compared with PFGE REA, RAPD analysis is faster, relatively simple and more economical.
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ObjectiveTodetectM.genitalium(Mg),M.penetrans(Mpe),M.pirum(Mpi),M.fermentans(Mf),Ureaplasmaurealyticum(Uu)andM.hominis(Mh)infectionsinurethra/cervicalcanalandpharynxandexploretheirclinicalsignificance.MethodsCultureandPCRwereperformedin72patientswithNGU/MPCtodetectMg,Mpe,Mpi,Mf,UuandMh.Thesecretionsfromurethra/cervicalcanalandpharynxweretested.ResultsMg,Mpe,Mpi,Mf,UuandMhweredetectedfromgenitalspecimensin23.6%,12.5%,2.8%,0,26.4%and8.5%ofpatients,respectively.Mg,Mpe,Mpi,Mf,UuandMhweredetectedfrompha-ryngealspecimensin24.6%,14.5%,0,0,2.9%and2.9%ofpatients,respectively.Thesamespeciesofmy-coplasmaswerefoundinbothgenitalandpharyngealspecimensin10patients(14.5%).ConclusionsUuandMginfectionsarecommoninpatientswithNGU/MPC.ThenewmycoplasmaspeciesMpeshouldbepaidattentionto.TheresultsindicatethatMgandMpemaybetransmittedbygenital-genitalsexandoral-genitalsex.MfmaybeofnoassociationwithNGU.
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Objective To detect and type herpes simplex virus (HSV) in genital lesions of the patients attending STD clinic. Methods Clinical data were collected and analyzed from patients with anogenital non-herpetic lesions including induration or furuncle, fissure, folliculitis, single ulcer and so on. HSV was detected and typed by culture and PCR with specimens taken from these lesions. Results One hundred and five cases were recruited in this study. Among them, 18 cases presented induration (furuncle), 15 fissure, 16 folliculitis, 7 abrasion, 12 single ulcer, 25 nonspecific erythema and 12 balanoposthitis with edema and exudation. HSV was found in 33.3%(6/18), 20%(3/15), 37.5%(6/16), 28.6%(2/7), 33.3%(4/12), 20%(5/25) and 50%(6/12) of these lesions, repectively, by PCR, while in 22.2%(4/18), 13.3%(2/15), 25%(4/16), 14.3%(1/7), 33.3%(4/12), 8%(2/25) and 41.7%(5/12), repectively, by viral culture. The positive rates of HSV from all these non-herpetic lesions were 30.5% (32/105) and 21% (22/105), respectively (? = 0.095, P = 0.114), by PCR and viral culture. The results of HSV typing were consistent between PCR and immunofluorescence with type-specific monoclonal antibodies. Among those with HSV infections, HSV-1 infection acounted for 9.4% (3/32), and HSV-2 90.6% (29/32). Conclusions The clinical manifestations of genital HSV infections vary, and HSV could be isolated from lesions of induration (furuncle), fissure, folliculitis, abrasion, single ulcer, nonspecific erythema and balanoposthitis with edema and exudation. HSV-2 is the predominant type.
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Objective To monitor antibiotic resistance of Neisseria gonorrhoeae in Nanjing City based on analysis of the results from 1999 to 2002. Methods The production of ?-lactamase was determined by paper acidometric method. Minimum inhibitory concentrations (MICs) of penicillin, ceftriaxone, tetracycline, ciprofloxacin and spectinomycin were determined by an agar plate dilution method. Results A total of 417 clinical isolates of Neisseria gonorrhoeae were examined. During the period of 1999-2002, positive rate of PPNG rapidly increased from 8.0% to 31.31% (P