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.

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.

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.

Objective To investigate DNA methylation markers in the whole blood of patients with systemic lupus erythematosus(SLE),in hope to facilitate the evaluation of SLE severity.Methods Whole blood samples were obtained from 58 patients with SLE(including 14 cases of severe SLE,25 moderate SLE,19 inactive SLE)and 50 healthy controls.Bisulphite sequencing was performed to determine the methylation status of interleukin-2 common receptor gamma chain(IL-2RG)promoter region,and real-time reverse transcriptionPCR to quantify the expression level of IL-2RG mRNA,in these subjects.Results The methylation level of IL2RG promoter region was 0.217 ± 0.140,0.325 ± 0.230,0.342 ± 0.085 and 0.175 ± 0.036 in the patients withsevere,moderate and inactive SLE and healthy controls,respectively.A significant increase was observed in the methylation level of IL-2RG promoter region in the patients with inactive SLE compared with the patients with severe SLE and healthy controls(both P ＜ 0.01),and in the patients with SLE compared with the healthy controls(0.263 ± 0.047 vs.0.175 ± 0.036,P ＜ 0.05).The expression level of IL-2RG mRNA was significantly lower in the patients with SLE than in the healthy controls(2.550 ± 0.823 vs.4.293 ± 1.283,P ＜ 0.05).A negative correlation was observed between the expression level of IL-2RG mRNA and methylation level of IL2RG promoter region in 20 patients with SLE(r =-0.44,P ＜ 0.05).Conclusion The methylation status of IL2RG promoter region is statistically higher in patients with SLE than in healthy controls,and significantly different between patients with active SLE and those with stable SLE.

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.

Objective: To observe the clinical efficacy and safety of Yiqi Tongluo Decoction in the intervention of early traditional Chinese medicine (TCM) syndromes after ischemic cerebrovascular disease (ICVD) intervention. Methods: From October 2020 to July 2023, a randomized, double-blind, placebo-controlled study was conducted to include 60 patients with qi deficiency, blood stasis, and phlegm obstruction syndrome after ICVD interventional therapy. They were assigned to the Yiqi Tongluo Decoction treatment group (30 cases) and the TCM placebo routine treatment control group (30 cases) according to the randomized block design. Both groups received routine standardized treatment of Western medicine, including dual antiplatelet, lipid regulation, and control of risk factors for cerebrovascular disease. The treatment group was treated with Yiqi Tongluo Decoction based on the control group. The course of treatment was 60 days and follow-up was carried out 2 and 6 months after the operation. The improvement of qi deficiency syndrome, blood stasis syndrome, phlegm syndrome score and TCM syndrome score, modified Rankin score (mRS), Barthel index (BI) score, Fatty acid-binding protein 4 (FABP4) level, incidence of transient ischemic attack (TIA) and ischemic stroke (IS) and incidence of adverse reactions, Head and neck CT angiography (CTA) or digital subtraction angiography (DSA) examination were collected. The clinical efficacy of the patients 2 months after the operation was taken as the main outcome index to preliminarily evaluate the early and long-term efficacy of Yiqi Tongluo Decoction after the ICVD intervention. The early and long-term clinical efficacy and safety of Western medicine standardized treatment combined with TCM Yiqi Tongluo Decoction on patients with qi deficiency, blood stasis and phlegm obstruction syndrome after ICVD intervention were evaluated. The safety of Yiqi Tongluo Decoction in the treatment of patients after ICVD intervention with white blood cell (WBC), C-reactive protein (CRP), fibrinogen (FIB), plasminogen time (PT), recurrence of cerebral ischaemia and restenosis in patients at 2 and 6 months after treatment were evaluated. Results: Compared to the control group, the TCM syndrome scores for qi deficiency, blood stasis and phlegm syndrome in the treatment group reduced significantly, the clinical efficacy improved significantly, the mRS score and FABP4 were reduced, and the BI score was increased. Adverse events such as cerebral ischaemia were fewer in the treatment group than in the control group, but the difference was not statistically significant; levels of CRP, WBC and PT were reduced, and levels of FIB were reduced at 6 months post-treatment, all P<0.01, and images were intuitively compared. The treatment group was superior to the control group. Conclusion Yiqi Tongluo Decoction combined with Western medicine standard treatment can improve the early clinical efficacy of ICVD patients with qi deficiency, blood stasis and phlegm obstruction syndrome after interventional surgery, improve neurological impairment and daily living ability, reduce the state of qi deficiency syndrome, blood stasis syndrome and phlegm syndrome after interventional surgery, and improve the clinical efficacy of TCM. At the same time, it can reduce the level of FABP4, the target of atherosclerosis and restenosis after interventional surgery, reduce the level of inflammation after interventional surgery in patients with ICVD, regulate coagulation function, and reduce the incidence of long-term recurrence of cerebral ischemia after interventional surgery, with good safety.