Objective: To analyze the expression of sigma factor in drug resistance gene mutations of Mycobacterium tuberculosis (MTB), so as to provide a reference for the drug resistance mechanism of tuberculosis. Methods: Clinical sputum specimens of outpatients at Tianjin Center for Tuberculosis from 2018 to 2022 were collected. A total of 899 MTB-positive strains were obtained by culture, and 492 phenotypically sensitive strains and 407 phenotypically resistant strains were identified by an in vitro phenotypic drug susceptibility test. Thirty drug-sensitive strains of MTB were randomly selected, and 98 drug-resistant strains with specific resistance phenotypes were chosen; all were subjected to melting curve analysis for detection of drug-resistance gene mutations. The strains were divided into sensitive strains without gene mutation, isoniazid-resistant strains with inhA mutation or katG mutation, rifampicin-resistant strains with rpoB mutation, and multigene mutation-resistant strains with inhA+rpoB mutation or katG+rpoB mutation. The mRNA relative expression of sigma factor was detected by fluorescence quantitative PCR, and the ratio of sigma factor mRNA relative expression between the experimental strain and the standard strain >2 was used to screen for highly expressed sigma factor. The differences in sigma factor mRNA relative expression and high expression rate between drug-resistant gene mutant strains and sensitive strains were analyzed. Results: Thirty sensitive strains and 90 drug-resistant strains were included. Among them, there were 16 strains with inhA mutation, 22 strains with katG mutation, 13 strains with rpoB mutation, 15 strains with inhA+rpoB mutation, and 24 strains with katG+rpoB mutation. Compared to the sigma factors of the sensitive strains, the mRNA expression levels of sigG and sigI in inhA-mutated strains, sigF, sigG, sigH, sigI, sigJ, and sigL in katG-mutated strains, and sigF, sigG, sigH, sigJ, and sigL in rpoB-mutated, inhA+rpoB-mutated, and katG+rpoB-mutated strains were significantly higher (all P<0.05). Additionally, the high-expression rates of sigI in inhA-mutated strains, sigF, sigG, sigI, sigJ, and sigL in katG-mutated and inhA+rpoB-mutated strains, and sigF, sigG, sigH, sigJ, and sigL in rpoB-mutated and katG+rpoB-mutated strains were also higher (all P<0.05). Conclusion Compared to sensitive MTB strains, sigI showed higher relative expression of mRNA and high-expression rate in inhA-mutated strains, and sigF, sigG, sigJ, and sigL had higher mRNA relative expression and high-expression rates in katG-mutated, rpoB-mutated, and multi-drug-resistant strains.

Objective　To investigate whether the expression regulation of regulatory gene Sigma factors (sigA-sigM) is related to isoniazid resistance phenotype in isoniazid-resistant Mycobacterium tuberculosis (MTB) caused by katG mutations, and to provide reference for the study of the molecular mechanism of isoniazid resistance. Methods　A total of 90 strains were collected from the patients undergoing first-line treatment at the Tianjin Tuberculosis Control Center during drug resistance testing from 2020 to 2022, of which 30 strains were sensitive strains without katG mutation, and 65 strains were isoniazid-resistant strains caused by katG mutation, including 11 strains resistant only to isoniazid, 24 strains resistant to both isoniazid and streptomycin, and 30 strains multidrug-resistant to isoniazid, streptomycin, and rifampicin. After the strains were collected on the isoniazid drug-containing medium, the RNA of the strains was extracted, and the relative expression levels of Sigma factors were detected by reverse transcription-polymerase chain reaction(RT-PCR). The expression differences of Sigma factors in different isoniazid drug-resistant phenotypes were analyzed by the Mann-Whitney U test. Results The gene expression levels of sigA, sigC, sigF, sigG, sigH, sigI, sigJ, sigK, sigL in isoniazid mono-resistant group were significantly higher than those in pan-isoniazid-sensitive group (Z=4.368, 5.701, 6.865, 4.021, 5.126, 2.670, 5.983, 4.701, 5.490, P all<0.001). The gene expression levels of sigA, sigC, sigF, sigG, sigH, sigI, sigJ, sigK, sigL in poly-resistance group were significantly higher than those in pan-sensitivity group (Z=-5.017, -4.670, -4.667, -5.456, -4.083, -5.393, -4.712, -6.971, -8.206, -5.211, P all<0.001). In multidrug-resistant (MDR) group, the gene expression levels of sigC, sigD, sigE, sigF, sigG, sigH, sigI, sigJ, sigK, sigL were significantly higher than those in pan-isoniazid-sensitive group (Z=-5.537, -4.003, -5.216, -7.328, -7.730, -5.658, -4.440, -6.036, -4.862, -4.312, P all<0.001). The expression levels of sigB, sigF, sigG showed statistically significant differences in gene expression between the isoniazid mono-resistant, isoniazid poly-resistant, and isoniazid multidrug-resistant groups (Z=10.139, 7.735, 14.532, P all<0.001). The expression rates of sigF, sigG, sigI, sigJ, and sigL in the isoniazid-resistant group were significantly higher than those in isoniazid sensitive group (χ2=17.410. 45.673. 57.661. 42.896. 26.363, P all<0.001). Conclusions sigF, sigG, sigI, sigJ, and sigL are associated with isoniazid resistance due to katG mutations.

Objective:To analyze the risk factors of neonatal nosocomial infection and establish a risk prediction model.Methods:The medical records of patients in the neonatal ward of Shengjing Hospital of China Medical University in 2023 were collected and divided into infection group and non-infection group according to whether nosocomial infection occurred. Univariate analysis was used to compare the differences of related factors between two groups. Multivariate Logistic regression analysis was used to screen the independent risk factors of neonatal nosocomial infection and construct the risk prediction model. The predictive value of the model was analyzed by receiver operating characteristic curve.Results:A total of 3 487 neonates were included in the study, including 1 881 males and 1 606 females. The median gestational age was 35 (32,38) weeks, and the median birth weight was 2 470(1 760,3 200) g. There were 158 cases in infection group and 3 329 cases in non-infection group. The main types of nosocomial infection were neonatal septicemia (142 cases,85.5%), central venous catheter-related infection (13 cases, 7.8%) and respiratory tract infection (eight cases, 4.8%). The main pathogens were Klebsiella pneumoniae (50 strains, 29.8%), Escherichia coli (26 strains, 15.5%)，and Staphylococcus epidermidis (25 strains, 14.9%). The independent risk factors of neonatal nosocomial infection were low birth weight ( OR=0.999,95% CI 0.999-1.000, P=0.039), central venous catheter placement ( OR=2.122, 95% CI 1.359-3.315, P=0.001), tracheal intubation ( OR=3.241, 95% CI 1.811-5.798, P<0.001)，and parenteral nutrition ( OR=11.087,95% CI 3.696-33.258, P<0.001). The regression equation of the established risk prediction model was Logit( P) =-47.407-0.001× birth weight +0.752× central venous catheter placement +1.176× tracheal intubation +2.406× parenteral nutrition. The area under the receiver operating characteristic curve was 0.870, the sensitivity was 89.2%, and the specificity was 76.0%,respectively. Conclusion:The risk prediction model of low birth weight, central venous catheter placement, tracheal intubation and parenteral nutrition is a good predictor of neonatal nosocomial infection and has certain clinical application value.

Objective:To explore the clinical efficacy and safety analysis of a novel laser localization technology assisted percutaneous puncture of trigeminal nerve microsphere capsule compression surgery for the treatment of primary trigeminal neuralgia.Methods:A retrospective selection was conducted on 63 patients with primary trigeminal neuralgia who underwent percutaneous puncture of the trigeminal nerve microsphere capsule compression surgery at the First Hospital of Hunan University of Chinese Medicine from January 2020 to December 2021. According to different surgical methods, they were divided into a new laser localization assisted puncture group (observation group) of 32 cases and a traditional barehanded localization puncture group (control group) of 31 cases. An analysis was conducted on the surgical time, puncture time, puncture frequency, intraoperative exposure to radiation, number of cases of poor balloon formation, and clinical efficacy within 6 months after surgery for two groups of patients. The prognosis of the patients was followed up at 6 months after surgery.Results:The surgical time, puncture time, puncture frequency, and intraoperative exposure of the observation group were all less than those of the control group, and the differences were statistically significant (all P<0.05). There was no statistically significant difference ( P>0.05) in the number of cases of poor balloon angioplasty between the observation group and the control group, as well as the pain score grading of the Barlow Neurological Institute (BNI) on the first day after surgery. Within 6 months after surgery, there was no statistically significant difference in the incidence of facial numbness, diplopia, masseter weakness, perilabial herpes, and recurrent pain between the two groups of patients (all P>0.05). Conclusions:Laser positioning technology can assist in precise puncture of the foramen ovale and accurate placement of balloons based on surgical experience, which helps to improve surgical safety, reduce postoperative complications and intraoperative radiation dose, and achieve satisfactory short-term follow-up results.

ObjectiveTo establish a vaginal self-sampling HPV cervical cancer screening model in Hainan Province, to analyze the application of p16 protein detection in HPV positive and non-HPV16 /18 shunt screening. MethodsFrom January 2019 to September 2022, a total of 200 women from the targeted population was randomly selected for vaginal self-sampling HPV typing test to screen cervical cancer using randomized numeric table method, followed by cervical cytology sampling for cytology p16 protein detection. Postoperative pathological examination was used as the gold standard. Multivariate logistic regression analysis was used to analyze the influencing factors of HPV positive detection rate in cervical lesions, and the nomogram model was constructed simultaneously. The receiver operating characteristic(ROC) curve and calibration curve were used for evaluating the accuracy of the nomogram model. Differences in the distribution of self-sampled HPV-positive and HPV infected genotypes were recorded, and the application of p16 protein detection in HPV-positive and non-HPV16/18 shunt screening was analyzed. ResultsAged ≥40 years, BMI ≥28.00 kg·m^-2, number of sexual partners ≥2, frequency of sexual life ≥10 times·month^-1, bleeding from sexual intercourse, and age of first sexual intercourse <22 years were the risk factors for HPV positive of cervical lesions (all P<0.001). The results of ROC curve and calibration curve showed that the area under ROC curve (AUC) was 0.874 (95%CI: 0.823‒0.907, P<0.05), the sensitivity was 0.835, the specificity was 0.847, and the Youden index was 0.672, indicating a good fit of the model. Results of vaginal self-sampling HPV test showed that the positive rate of HPV was 86.50% (173/200). HPV high-risk infection types mainly included HPV16, 18, 31, 33, 35, 39, 45, 52, 56, 58, 59, 68, 73, and 82. Single HPV infection accounted for 95.95% (166/173), 2.89% (5/173) were infected with two types of HPV, and 1.16% (2/173) were infected with three or more types of HPV. Colposcopic pathologic diagnosis was used as the gold standard, and the results showed that the accuracy of p16 protein detection in the diagnosis of cervical cancer was 93.50% (187/200), with a sensitivity of 96.53% (167/173), and a specificity of 74.07% (20/27). The negative and positive predictive value were 76.92% (20/26) and 95.98% (167/174), respectively. The results of shunt screening showed that there were 80 cases infected with HPV16, 79 cases infected with HPV18 and 41 cases of non-HPV16/18, with a sensitivity of 90.91%, 90.32% and 86.67%, a specificity of 71.43%, 64.71% and 72.73%, a negative predictive value of 62.50%, 64.71% and 66.67%, a positive predictive value of 93.75%, 90.32% and 89.66%, and an accuracy of 87.50%, 84.81% and 82.93%, respectively. The specificity and accuracy of p16 positive screening for cervical cancer were significantly higher than that of HPV positive detection, but the false positive rate was significantly lower than that of HPV positive detection. The AUCs of HPV positive, p16 positive and combination of the two detection methods for cervical cancer were 0.603, 0.822 and 0.907, respectively. ConclusionVaginal self-sampling HPV testing is a widely accepted mode for cervical cancer screening. Cervical cytology p16 protein detection is important for self-sampled HPV positive and shunt screening of non-HPV16/18.

Objective:To explore the sleep status and impact factor analysis of patients in clinical trials of antineoplastic drugs, and provide a basis for improving the sleep status and impact analysis of patients in clinical trials of antineoplastic drugs.Methods:From April to May 2023, 107 oncology patients in the Phase I Clinical Trial Ward of the Affiliated East Hospital of Tongji University were selected as the research objects by convenient sampling method. The general information questionnaire, Pittsburgh Sleep Quality Index Scale (PSQI), Numeric rating scale (NRS), Generalized Anxiety Disorder Scale (GAD-7) and Depression Self-Ration Tool Scale (PHQ-9). Multivariate Logistic regression analysis methods were used to carry out a cross-sectional investigation and the relevant factors affecting patients′sleep.Results:Totally 103 questionnaires were effectively collected. The 103 patients′ age ranged from 20 to 75 years old, including 61 males and 42 females. 47.57% (49/103) patients in clinical trials of antineoplastic drugs had abnormal sleep. The average score of patients (PSQI) (7.66 ± 3.93) was higher than the average score of the domestic norm (3.88 ± 2.52), and there was significant statistical difference ( t = 9.76, P<0.01). Logistic regression analysis showed that pain ( OR = 3.004, 95% CI 1.135-7.948, P<0.05) and trial cycle ( OR = 0.432, 95% CI 0.191-0.978, P<0.05) were significant risk factors for abnormal sleep quality. Conclusions:The incidence of abnormal sleep quality in patients of clinical trials of antineoplastic drugs is high, but the sleep quality is poor. The factors that affect the sleep quality of patients in clinical trials of antineoplastic drugs are mainly related to the patient′s trial cycle and cancer pain. According to these characteristics, individualized programs should be developed to improve the sleep quality of patients with advanced cancer, so as to improve the quality of life of patients with advanced cancer.

Objective　To investigate the relationship between the degree of cervical lesions and the status of paired box-1 (PAX1) and tumor protein 63(TP63) gene promoter methylation in human papillomavirus (HPV)-positive patients with cervical lesions, as well as analyze their clinical significance. Methods Cervical tissue specimens were collected from 128 patients who were suspected of cervical lesions and HPV infection, and admitted to Qionghai People's Hospital between December 2021 and December 2022. According to pathological examination results, the patients were divided into the low-grade squamous intraepithelial lesion (LSIL) group (n=43), high-grade squamous intraepithelial lesion (HSIL) group (n=51) and cervical cancer group (n=34). The second-generation hybrid capture method was used for viral load. The degree of PAX1 and TP63 gene promoter methylation in each group was detected by bisulfite sequencing, and mRNA expression of PAX1 and TP63 was detected by real-time quantitative PCR. The diagnostic performance of the degree of PAX1 and TP63 methylation for cervical intraepithelial neoplasia (CIN2+) was evaluated. Results There were statistically significant differences in HR-HPV viral load between the groups (P>0.05). A total of 49 (38.28%) patients with PAX1 gene promoter methylation, and 55 (42.97%) patients with TP63 gene promoter methylation were detected among the 128 patients. The percentages of PAX1 and TP63 gene promoter methylation in the cervical cancer group, HSIL group and LSIL group were (67.65% and 73.53%), (43.14% and 49.02%) and (9.30% and 11.63%), with statistically significant differences between groups (P<0.05). The mRNA expression levels of PAX1 and TP63 in the cervical cancer group, HSIL group and LSIL group were [(0.34±0.08) and (0.45±0.13)], [(0.72±0.11) and (0.63±0.09)], [(1.04±0.09) and (0.87±0.11)], with statistically significant differences between groups (P<0.05). Receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) values of PAX1, TP63 gene promoter methylation, and their combination for diagnosing CIN2+ were 0.793, 0.842, and 0.857, respectively. The sensitivity values were 77.78%, 83.33%, and 77.78%. The specificity values were 80.85%, 85.11%, and 93.62%. The combined detection can improve the specificity of diagnosis of CIN2+ lesions. Conclusions The degree of PAX1 and TP63 gene promoter methylation is closely related to cervical lesions in patients with HPV infection, which indicates that it can be used as potential auxiliary indicators for the clinical diagnosis of CIN2+lesions.

Objective:To investigate the distribution of respiratory pathogens and risk factors of death in patients with pulmonary infection in neurosurgical intensive care unit (NICU).Methods:A total of 87 patients with pulmonary infection in the NICU of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine from January 2018 to December 2019 were collected, and the pathogens of their respiratory tract were analyzed to understand the types and distribution of bacteria in the lung infection. Univariate statistical analysis was used to analyze the relationship between the patient′s clinical outcome with age, diabetes, hypertension, renal insufficiency, hypoproteinemia, anemia, chronic respiratory disease, surgery, tracheotomy, and bacterial multi-resistance. Binary logistic regression analysis was used to analyze the influencing factors of death in NICU patients with pulmonary infection.Results:A total of 112 pathogenic bacteria were isolated in this research group, including 83 Gram-negative bacteria (74.11%), 22 Gram-positive bacteria (19.64%), and 7 Fungi (5.25%). Imipenem was highly sensitive to Gram-negative bacteria, vancomycin was highly sensitive to Gram-positive bacteria, and other drugs were highly resistant. 41 patients died (47.13%). Age≥60 ( OR=3.501, 95% CI: 1.152-10.638), renal insufficiency ( OR=3.872, 95% CI: 1.336-11.224), tracheotomy ( OR=0.317, 95% CI: 0.114-0.882), bacteria multi-drug resistance ( OR=3.480, 95% CI: 1.162-10.422) were independent risk factors for death in NICU patients with pulmonary infection. Conclusions:Patients with severe neurological diseases are in critical condition, and there are many patients with pulmonary infection, with poor prognosis and high mortality. Gram-negative bacteria are the most common respiratory pathogens. Carbapenems account for the highest proportion of antibiotics in clinic. Advanced age, renal insufficiency and bacterial multidrug resistance increase the mortality of patients, while early tracheotomy can reduce the mortality of patients.

Objective:To investigate the correlation between the number of circulating tumor cells (CTC) in peripheral blood and clinicopathological features of patients with breast cancer.Methods:The clinical data of 104 breast cancer patients at Guangzhou Panyu Central Hospital between January 2017 and May 2020 were retrospectively analyzed. The number of CTC in peripheral blood, the levels of serum tumor markers [alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen (CA)125, CA153] were detected. In blood samples, the number of CTC ≥ 2/ml was defined as CTC positive. Immunohistochemistry was used to analyze the protein expression of Ki-67 in tumor tissues. The association of CTC with clinicopathological features, serum tumor markers and Ki-67 protein expression was also analyzed.Results:The CTC positive rate was 80.77% (84/104). There were statistically significant differences in composition of whether there was vascular tumor thrombus (χ 2 = 0.860, P = 0.009), axillary lymph node metastasis (χ 2 = 12.382, P<0.01), N staging ( P = 0.002) and TNM staging (χ 2 = 7.698, P = 0.006) between patients with CTC positive and negative. However, there were no statistically significant differences in composition of age ( t = 0.634, P = 0.528), tumor quadrant (χ 2 = 6.523, P = 0.163), molecular subtyping (χ 2 = 4.164, P = 0.384), histological grade (χ 2 = 1.901, P = 0.387), T staging ( P = 0.099) and whether there was nerve invasion (χ 2 = 0.092, P = 0.761). The levels of serum CEA and CA125 in CTC positive patients were higher than those in CTC negative patients [median ( P25, P75): 2.50 ng/ml (2.21 ng/ml, 2.92 ng/ml) vs. 1.89 ng/ml (1.61 ng/ml, 2.35 ng/ml); 13.81 U/ml (11.79 U/ml, 16.28 U/ml) vs. 11.17 U/ml (8.91 U/ml, 12.80 U/ml); all P < 0.05], and CTC was positively correlated with serum CEA and CA153 levels ( r = 0.520, P<0.01; r = 0.497, P<0.01); CTC was not related to Ki-67 protein expression (χ 2 = 0.512, P = 0.474). Conclusion:The number of CTC in peripheral blood is closely related to clinical staging, lymph node or hematogenous metastasis, tumor markers CEA and CA153 levels of breast cancer. The increased number of CTC may cause tumor progression and metastasis.

Objective To evaluate transurethral fulguration with intravesical instillation of heparin and alkalinized lidocaine for the treatment of interstitial cystitis/bladder pain syndrome (IC/BPS).Methods Data from the chnical records of 31 patients (30 female and 1 male) with IC/BPS were analyzed retrospectively.Transurethral fulguration and biopsy were performed.Intravesical instillation of heparin and alkalinized lidocaine (25 000 units of hepmin,10 mL of 2％ lidocaine,and 5 mL of 5％ sodium bicarbonate) was administered weekly for 8 weeks.Thereafter,intmvesical instillation treatment was administered twice a month.The interstitial cystitis symptom index and problem index (O'Leary-Sant index),visual analog scale score for pain,quality-of-life index,voiding frequency,bladder capacity,and side effects of intravesical instillation were recorded preoperatively and at the first and sixth month follow-ups postoperatively.Results The follow-up period was 6 to 24 months.The interstitial cystitis symptom index and problem index,visual analog scale score for pain,quality-of-life index,daily voiding time,and maximal bladder volume improved significantly in 28 cases (90.32％;P ＜ 0.01),and no significant adverse effects were observed.Two patients underwent cystectomy,and the symptoms disappeared after the operation.Carcinoma in situ was detected on histopathological examination in one patient.Conclusion Transurethral fulguration with intravesical instillation of heparin and alkalinized lidocaine is a safe and effective therapy for IC/BPS.