Increased risk of end-stage renal disease in patients with systemic sclerosis.

OBJECTIVE: Systemic sclerosis (SSc) is a systemic autoimmune disease affecting multiple organs, including the kidneys. There is a lack of long-term renal prognosis studies on patients with SSc. The aim of this study was to assess the risk of end-stage renal disease (ESRD) in patients with SSc. METHOD: We designed a prospective cohort study based on the National Health Insurance Research Database of Taiwan. Patients with SSc and a non-SSc control group were selected from 1 January 2000 to 31 December 2013. The SSc cohort and control group were matched on the propensity score in a 1:2 ratio. The primary outcome was development of ESRD. Cox proportional hazard regression was performed to assess the effects of SSc on ESRD. RESULTS: After propensity score matching, we enrolled 2012 patients in the SSc group and 4024 patients in the control group. During a mean follow-up of 6.5 years, 86 individuals [SSc group, n = 41 (2.04%); control group, n = 45 (1.12%)] had developed ESRD. The risk of ESRD in the SSc group was approximately two times higher than that in the control group [hazard ratio (HR) = 2.12, 95% confidence interval (CI) 1.39-3.24]. Subgroup analysis revealed that the higher risk of ESRD was predominantly in males (HR = 4.14, 95% CI 1.97-8.71) and the younger population (HR = 7.09, 95% CI 2.31-21.80). CONCLUSION: There was a significantly higher risk of ESRD among SSc patients than among the general population, with males and younger generations being the most vulnerable groups.

[Association of DNA methylation of IFNG gene with no/low response to hepatitis B vaccine in children].

Objective: To explore the association of DNA methylation with immune response to hepatitis B (HepB) vaccine in Han nationality children from Guangxi province. Methods: A total of 263 children aged 8-9 months who had completed HepB immunization program were recruited from three hospitals in Guangxi province by using unmatched case-control method. Children with the HepB surface antibody concentration(Anti-HBs)<100 mIU/ml was set as the case group and ≥100 mIU/ml as the control group. Multiplex PCR and heavy sulfite sequencing were used to treat the samples. Illumina platform was used for high-throughput DNA methylation sequencing of IFNG gene target regions and CpG sites. Unconditional logistic regression was used to analyze the association between cytosine-phospho-guanosine DNA methylation at 18 loci of IFNG gene and HepB immune response level. Results: There were 104 children in the case group and 159 in the control group. The median (Q1, Q3) level of anti-HBs in two groups were 62.34 (30.06, 98.88) mIU/ml and 1 089.10 (710.35, 1 233.45) mIU/ml. The methylation levels of IFNG_1 gene 44 and 93 locus in the case group were higher than those in the control group (P<0.05). The unconditional logistic regression model showed that the DNA methylation level of IFNG_1 gene at 44 (OR=1.18, 95%CI: 1.03-1.35) and 93 (OR=1.21, 95%CI: 1.07-1.38) locus was associated with the HepB response level. Conclusion: The changes of DNA methylation at locus 44 and 93 of IFNG_1 gene may be relevant factors affecting the response level of HepB in Han nationality children from Guangxi province.

[Application of artificial intelligence-assisted diagnosis for cervical liquid-based thin-layer cytology].

Objective: To explore the application value of artificial intelligence-assisted diagnosis system for TBS report in cervical cancer screening. Methods: A total of 16 317 clinical samples and related data of cervical liquid-based thin-layer cell smears, which were obtained from July 2020 to September 2020, were collected from Southern Hospital, Guangzhou Huayin Medical Inspection Center, Shenzhen Bao'an People's Hospital(Group) and Changsha Yuan'an Biotechnology Co., Ltd. The TBS report artificial intelligence-assisted diagnosis system of cervical liquid-based thin-layer cytology jointly developed by Southern Medical University and Guangzhou F. Q. PATHOTECH Co., Ltd. based on deep learning convolution neural network was used to diagnose all clinical samples. The sensitivity,specificity and accuracy of both artificial intelligence-assisted diagnosis system and cytologists using artificial intelligence-assisted diagnosis system were analyzed based on the evaluation standard(2014 TBS). The time spent by the two methods was also compared. Results: The sensitivity of artificial intelligence-assisted diagnosis system in predicting cervical intraepithelial lesions and other lesions (including endometrial cells detected in women over 45 years old and infectious lesions) under different production methods, different cytoplasmic staining and different scanning instruments was 92.90% and 83.55% respectively, and the specificity of negative samples was 87.02%, while that of cytologists using artificial intelligence-assisted diagnosis system was 99.34%, 97.79% and 99.10%, respectively. Moreover, cytologists using artificial intelligence-assisted diagnosis system could save about 6 times of reading time than manual. Conclusions: Artificial intelligence-assisted diagnosis system for TBS report of cervical liquid-based thin-layer cytology has the advantages of high sensitivity, high specificity and strong generalization. Cytologists can significantly improve the accuracy and work efficiency of reading smears by using artificial intelligence-assisted diagnosis system.

[Clinical features of 9 cases of Brucella endocarditis].

Objective: To explore the clinical characteristics of patients with Brucella endocarditis. Methods: The clinical data of 9 patients with Brucella endocarditis admitted to Beijing Ditan Hospital from October 2008 to August 2018 were retrospectively analyzed. Through the electronic medical record system of the hospital. Through assessing the electronic medical record system of the hospital, demographic data, main symptoms, vital signs, blood culture, Rose Bengal Plate Agglutination Test, echocardiography, electrocardiogram, chest imaging and other clinical data of included patients were inquired and recorded. Patients were followed up by telephone for medication, operation and outcome. Results: The 9 patients were all Han nationality, aged from 25 to 66 years, 7 out of 9 patients were male, and they came from Hebei, Shandong, Shanxi, Inner Mongolia and Beijing. Of the 9 patients, 5 were farmers, 2 were self-employed, 1 was a technician, and 1 was unemployed. Of the 9 cases, 8 had a history of close contact with cattle and sheep, and 5 had a history of eating beef and mutton. Rose-Bengal Plate Agglutination Test and blood culture were positive in all 9 patients. Aortic valve was involved in 7 out of 9 patients, mitral and tricuspid valve was involved in 1 patient, respectively, and aortic dissection occurred in 1 patient. Condition of 1 patient rapidly deteriorated after admission and finally died during hospitalization despite antibiotic therapy, the remaining patients received long-term antibiotic treatment. A total of 7 patients who underwent valve replacement were followed up. One patient died of cerebral hemorrhage 6 months after operation, and the remaining 6 patients recovered well after valve replacement. Heart failure occurred in all 9 patients, and pericardial effusion occurred in 8 patients. Electrocardiogram showed low voltage of the QRS complex in the limb in 3 cases and poor R-wave progression in V(1)-V(3) lead in 2 cases, and sinus tachycardia in 2 cases. One patient developed non-specific ST-T abnormalities. All patients had fever, 7 patients complained of weakness, and 6 patients complained of palpitations. Among the 9 patients, 7 cases had anemia, 7 patients had pneumonia, 6 had bilateral pleural effusion, 4 had thrombocytopenia. Creatinine was above normal in 4 patients, urine protein was positive in 3 patients, Delta Bilirubin was higher in 3 cases. Conclusions: Patients with Brucella endocarditis often suffer from heart failure and have severe complications. Adequate antibiotic therapy in combination with valve replacement is effective for the treatment of patients with Brucella endocarditis.

Tetramethylpyrazine (TMPZ) triggers S-phase arrest and mitochondria-dependent apoptosis in lung cancer cells.

Tetramethylpyrazine (TMPZ) is one of the active compounds extracted from the traditional Chinese herb Chuanxiong. Several studies have shown its anti-cancer properties. However, its functions in lung cancer and the underlying cellular mechanisms are relatively unknown. Our present study aimed to investigate the effects of TMPZ on A549 and 95D cells. The MTT assay showed that TMPZ decreased cell viability in a dose- and time-dependent manner. The results of the colony formation assay indicated that TMPZ strongly suppressed colony formation ability in A549 and 95D cells. Flow cytometric analysis revealed that TMPZ induced S phase arrest in lung cancer cells. In addition, TMPZ induced apoptosis, as shown by the results of propidium iodide/Annexin V double-staining. Furthermore, TMPZ decreased mitochondrial membrane potential (∆Ψm) in a dose-dependent manner. Finally, western blot analysis of TMPZ-treated cells revealed the activation of Caspase-3 and the increase of the ratio of Bax/Bcl-2. These results demonstrated that TMPZ could suppress carcinogenesis of lung cancer cells through blocking cell cycle and inducing mitochondria-dependent apoptosis by regulating Caspase-3 and Bax/Bcl-2, suggesting that TMPZ may be a promising drug to treat lung cancer.

Genetic Polymorphisms and Mutations of 30 Y-STR Loci in Chinese Han Population.

OBJECTIVES: To investigate the genetic polymorphisms and mutations of 30 Y-STR loci in Chinese Han males and to evaluate its forensic application. METHODS: The DNA extracted from blood samples of 1 005 unrelated males and 1 008 father-son pairs （1 949 individuals in all） in Chinese Han population were typed using developed 30 Y-STR loci identification system. The parameters of population genetics and the mutation rates of each locus were analysed statistically. RESULTS: A total of 983 haplotypes were found in 1 005 unrelated males from Chinese Han population, of which 963 were unique. The overall haplotype diversity （HD） and discrimination capacity （DC） were 0.999 955 and 0.978 109, respectively. Totally 340 alleles were detected on 30 Y-STR loci, the value of gene diversity （GD） ranged from 0.410 3 to 0.952 3. The GD values of 24 out of the 30 loci were over 0.6. There were 30 269 allele transfers in 1 008 father-son pairs, one mutation in 68 father-son pairs, and the mutation of three father-son pairs occurred at two loci. On 26 Y-STR loci, 74 mutations were detected in 71 father-son pairs. The average mutation rates were 2.4×10⁻³ （95% CI： 1.9×10⁻³-3.1×10⁻³ï¼‰. Seventy-three mutation events were one-step mutation （98.6%）, 1 mutation event was two-step mutation （1.4%）. CONCLUSIONS: The multiplex PCR system with 30 Y-STR loci has high genetic polymorphism and low mutation rates in Chinese Han males. Therefore, the system shows important values in Y-STR database construction and population genetic research.

Periodically Modulated Size-Dependent Elastic Properties of Armchair Graphene Nanoribbons.

First-principles calculations were conducted on armchair graphene nanoribbons (AGNRs) to simulate the elastic behavior of AGNRs with hydrogen-terminated and bare edges. The results show width-dependent elastic properties with a periodicity of three, which depends on the nature of edge. The edge eigenstress and eigendisplacement models are able to predict the width-dependent nominal Young's modulus and Poisson's ratio, while the Clar structure explains the crucial role of edges in the periodically modulated size-dependent elastic properties.

Phylogeography of Cephalotaxus oliveri (Cephalotaxaceae) in relation to habitat heterogeneity, physical barriers and the uplift of the Yungui Plateau.

Habitat heterogeneity, physical barriers, and the uplift of the Yungui Plateau were found to deeply affect the phylogeographic pattern and evolutionary history of Cephalotaxus oliveri, a perennial conifer endemic to China. In this study, we explored the phylogeography using three chloroplast sequences (trnL-trnF, trnT-trnD and atpB-rbcL) in 22 natural populations of C. oliveri distributed throughout its range. The Yungui Plateau populations of C. oliveri were revealed to origin ca. 9.15Ma by molecular clock estimation, which is consistent with rapid uplift of the Qinghai-Tibetan Plateau (QTP) ca. 8-10Ma. Additionally, geological effects of the Yungui Plateau were suggested to promote the rapid intra-specific differentiation of C. oliveri in the Pliocene and Early Pleistocene. The relatively low level of genetic diversity (h=0.719, θ=1.17×10(-3)) and high population differentiation (NST=0.771 and GST=0.642) implied restricted gene flow among populations, which was confirmed by the Nested Clade Analysis (NCA). Mismatch distribution and haplotypes network provided evidences of recent demographic population expansion. Furthermore, the statistical dispersal-vicariance analysis indicated that the center of origin was in Central China. The comparison of haplotype distribution patterns indicated that the regions of HNHPS and HBLD were the potential refugia during the Pleistocene ice ages. Our results highlighted that habitat heterogeneity and physical barriers presenting in a species range can predict genetic patterns.

Association of chicken growth hormone polymorphisms with egg production.

Growth hormone (GH) has diverse functions in animals, together with other hormones from the somatotropic axis. Here, chicken GH (cGH) was investigated in recessive white chickens and Qingyuan partridge chickens as a candidate gene affecting egg production traits. Chicken egg production traits were studied in association with 4 selected single nucleotide polymorphisms (T185G, G662A, T3094C, and C3199T). Genotyping was performed by the polymerase chain reaction-ligase detection reaction method. T185G was significantly associated with the egg production traits of body weight at first egg (BW), egg weight at first egg (EW), and the total egg production of 300-day old birds (EN 300). T3094C was also significantly associated with certain egg production traits; however, it affected the 2 breeds differently. Haplotypes of the 4 single nucleotide polymorphisms were also significantly associated with egg production traits of chicken age at first egg laying, BW, EW, and EN 300. H1H6 was the most advantageous diplotype for egg production. We putatively concluded that polymorphisms in the cGH gene and its haplotypes could be used as potential molecular markers for egg production traits to enhance the breeding programs of indigenous chickens.

Is sudden deafness in non-diabetic patients affected by their glycosylated hemoglobin levels?

OBJECTIVE: Though the incidence of sudden sensorineural hearing loss (SSNHL) is relatively low, the disorder has a major impact on the quality of life of patients. Identifying biological markers for the disease will be useful, especially in resource-scarce areas. Our study aims to evaluate the correlation between the degree of hearing impairment and glycosylated hemoglobin (HbA1c) in patients with SSNHL. PATIENTS AND METHODS: One hundred and thirty-eight patients with SSNHL and no history of diabetes were included in this study. The intravenous HbA1c content before treatment was correlated with the pure tone audiogram (PTA) average as per the criteria for SSNHL. Spearman correlation and the Receiver Operating Characteristic (ROC) curve were used to determine the HbA1c levels of the study participants. The critical value of HbA1c and its diagnostic implications for assessing the degree of hearing impairment in patients with SSNHL were noted. RESULTS: There was a significant positive correlation between HbA1c and PTA in patients with SSNHL (p<0.05). In addition, the best HbA1c cutoff value for screening and referring an individual for a detailed audiometric evaluation of hearing impairment was 5.550%, as indicated by the ROC curve. CONCLUSIONS: The level of HbA1c in the circulation may affect the onset, duration, and progression of SSNHL. The same parameter may be used as a diagnostic and prognostic indicator for this condition.

[Transoral robotic nasopharyngectomy for local recurrent nasopharyngeal carcinoma].

Objective: To investigate the safety, efficacy, locally control and survival results of transoral Da Vinci robotic surgery for salvage treatment of locally recurrent nasopharyngeal carcinoma. Methods: This retrospective study included 33 patients with locally recurrent nasopharyngeal carcinoma (stage rT1-2, partial rT3) underwent transoral Da Vinci robotic surgery between October 2017 and January 2020. There were 20 males and 11 females, with an average age of (47.9±10.5) years. The lesions were localized in nasopharyngeal cavity in 14 cases, with extending to parapharyngeal space in 6 cases and the floor of sphenoid sinus in 13 cases. Transnasal endoscopy was used to assist surgery if necessary. SPSS 25.0 statistical software was used for statistical analysis. Results: Transoral robotic nasopharyngectomy was successfully performed in all cases without conversion to open surgery, of which 13 cases were combined with transnasal endoscopic surgery. The average operation time was (126.2±30.0) min, ranging from 90 to 180 min. The postoperative pathological margin was R0 (31 cases) and R1 (2 cases), with no tumor residue. Complications of surgery mainly included symptoms of headache, nasal dryness and velopharyngeal insufficiency without nasopharyngeal hemorrhage. Follow-up time was from 3 to 54 months. One case had tumor recurrence 11 months after operation, 1 case had ipsilateral cervical lymph node metastasis 27 months after operation, 2 cases had distant metastasis and 1 case died of nasopharyngeal hemorrhage 3 months after operation. The 1-year, 2-year and 3-year overall survival rates were 97.0%, 96.0% and 92.9%, respectively and the local recurrence free rates were 97.0%, 95.7% and 91.7%, respectively. Conclusion: Transoral robotic nasopharyngectomy is safe and feasible for local recurrent nasopharyngeal carcinoma in selected patients, with higher local control rate and quality of life.

[Sleep quality and sleep disturbances in Chinese pregnant women: a multicenter cross-sectional study].

Objective: This study aims to investigate the sleep quality of pregnant women in Xuhui District, Shanghai, and the related factors of sleep disturbances during pregnancy. Methods: From February 2019 to February 2021, we used online integrated sleep questionnaire (including PSQI, BQ, ESS, AIS) in Shanghai Jiao Tong University School of Medicine Affiliated Sixth People's Hospital, The International Peace Maternity and Child Health Hospitals of China Welfare Institution, and Shanghai Eighth People's Hospital, to investigate the sleep quality across pregnancy. We also collected maternal physical examination results, childbearing history, sociodemographic, and other clinical data. The prevalences and related factors of various sleep disturbances in pregnant women were analyzed, including insufficient/excessive nighttime sleep, low sleep efficiency, difficulty falling asleep, poor sleep quality, insomnia, daytime sleepiness, and high risk of sleep-disordered breathing (SDB). Results: This study includes 1 898 cases in the first trimester (T1), 3 099 cases in the second trimester (T2), and 1 539 cases in the third trimester (T3). Poor sleep quality (38.6%), daytime sleepiness (mild 41.9%, moderate 17.7%, severe 2.1%), and suspicious insomnia (32.3%) are most prevalent among women in T1 (P<0.01). In comparison, short sleep time (2.7%), long sleep time (8.6%), difficulty falling asleep (12.2%), poor sleep efficiency (35.4%), very poor sleep quality (6.7%), clinical insomnia (21.8%), and high-risk SDB (6.4%) are most prevalent among women in T3 (P<0.05). During pregnancy, late gestation (OR=1.016, 95%CI: 1.006-1.025) and multiple induced/drug abortions (OR=1.329, 95%CI: 1.043-1.692) are risk factors for poor sleep quality (PSQI>5), while multiple full-term deliveries (OR=0.800, 95%CI: 0.675-0.949) is its protective factor. Advanced maternal age (OR=0.976, 95%CI: 0.956-0.997), multiple full-term deliveries (OR=0.808, 95%CI: 0.680-0.959), late gestation (OR=0.983, 95%CI: 0.974-0.992) and hypertension (OR=0.572, 95%CI: 0.401-0.814) are protective factors for daytime sleepiness (ESS>6). The high-risk pregnancy category (OR=9.312, 95%CI: 1.156-74.978) is a risk factor for insomnia (AIS≥4), while multiple full-term deliveries (OR=0.815, 95%CI: 0.691-0.961) is its protective factor. High BMI (OR=1.334, 95%CI: 1.270-1.402) and hypertension (OR=4.427, 95%CI: 2.539-7.719) are risk factors for high-risk SDB in pregnant women. Conclusions: The prevalences of various sleep disturbances are high throughout pregnancy. Noticeably, symptoms of maternal SDB develop along with pregnancy. Different types of sleep disturbances are associated with different factors. Women of high-risk pregnancy category, in late gestation, with high BMI, hypertension, a history of induced/drug abortion, or without a history of full-term delivery can be at high risk of sleep disturbances during pregnancy.

Discovery and insights from DSX mission's high-power VLF wave transmission experiments in the radiation belts.

Space weather phenomena can threaten space technologies. A hazard among these is the population of relativistic electrons in the Van Allen radiation belts. To reduce the threat, artificial processes can be introduced by transmitting very-low-frequency (VLF) waves into the belts. The resulting wave-particle interactions may deplete these harmful electrons. However, when transmitting VLF waves in space plasma, the antenna, plasma, and waves interact in a manner that is not well-understood. We conducted a series of VLF transmission experiments in the radiation belts and measured the power and radiation impedance under various frequencies and conditions. The results demonstrate the critical role played by the plasma-antenna-wave interaction around high-voltage space antennae and open the possibility to transmit high power in space. The physical insight obtained in this study can provide guidance to future high-power space-borne VLF transmitter developments, laboratory whistler-mode wave injection experiments, and the interpretation of various astrophysical and optical phenomena.

Age- and gender-related long-term renal outcome in patients with lupus nephritis.

OBJECTIVE: To investigate age- and gender-related long-term renal outcome in patients with lupus nephritis (LN). METHODS: This is a retrospective, chart review study of patients with LN at Chang Gung Memorial Hospital, Kaohsiung, between January 1986 and June 2004. All had undergone a renal biopsy that showed LN. The end point of outcome was chronic renal insufficiency (CRI), which was defined as 'doubling of baseline serum creatinine lasting for at least 6 months with a value at least 2 mg/dl.' The patients were categorized by age tertiles and gender. A 5-year survival curve was constructed to study the effect of age and gender on the outcome. RESULTS: In total, 121 sets of patient data were evaluated. The study group included 104 women and 17 men. Of the study patients, 26 (21%) developed CRI after 5 years of follow-up. There was no significant difference among age groups in developing CRI (p = 0.23). In terms of gender, men had worse long-term renal outcome (p = 0.004) than women. CONCLUSIONS: The long-term renal outcome of the LN patients did not differ among age groups, but was worse in men.

[Effects of Freund's complete adjuvant on autophagy protein expression in rat tuberculous wound model].

Objective: To explore the effects of Freund's complete adjuvant on autophagy protein expression in rat tuberculous wound model. Methods: The experimental research method was used. In the first batch, twelve 6-week-old male Sprague-Dawley (SD) rats were sensitized by subcutaneous injection of Freund's complete adjuvant into the hips. Three weeks later, the rats were infected with attenuated Bacille Calmette-Guérin (BCG) subcutaneously on both sides of the back spine. After establishing the tuberculosis wound rat model, according to the random number table (the same grouping method below), the rats were divided into 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group, with 3 rats in each group, with continuous normal feeding to the corresponding days after infection. In the second batch, twenty-three 6-week-old male SD rats were divided into blank control group (n=3, normal feeding without any treatment), BCG alone group (n=5), BCG+ rapamycin group (n=6), BCG+ 3-methyladenine group (n=6), and BCG+ starvation group (n=3). The last 4 groups of rats were sensitized as before, and infected as before 1 week later. Rats in BCG alone group were fed normally without any treatment. Rats in BCG+ rapamycin group or BCG+ 3-methyladenine group were intraperitoneally injected with rapamycin or 3-methyladenine once every other day and fed normally. Rats in BCG+ starvation group were fasted for 48 hours after infection and then fed normally. All the rats in the first batch of 4 groups were sacrificed on the corresponding days after infection, and the tissue where the buttocks were injected with Freund's complete adjuvant was harvested; the tissue of rats in the second batch of BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were harvested the same as before 7 days after infection, and all the rats in blank control group were taken the same tissue at the same time point. Hematoxylin-eosin staining was performed to observe the structure and morphology of cells in the tissue harvested; immunohistochemistry was used to observe the protein expressions of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B) in the tissue harvested. Data were statistically analyzed with Kruskal-Wallis test and Bonferroni correction. Results: Inflammatory cell infiltration was observed in the tissue of rats where the Freund's complete adjuvant was injected in 8 d infection group, granuloma formation was seen in 15 d infection group, part of tissue cell necrosis was seen in 32 d infection group and 43 d infection group, and cell necrosis in 43 d infection group was worse than that in 32 d infection group. Seven days after infection, inflammatory cell infiltration was seen in the tissue of rats where the Freund's complete adjuvant was injected in BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group, while regular arrangement of cells and no inflammatory cell infiltration were observed in blank control group. There were no statistically significant differences in the protein expressions of Beclin-1 or LC3B in the tissue of rats where the Freund's complete adjuvant was injected in 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group (H=1.923, 5.821, P>0.05). Seven days after infection, the protein expressions of Beclin-1 and LC3B in the tissue of rats where the Freund's complete adjuvant was injected in blank control group, BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were respectively 0.325% (0.250%, 0.360%), 3.225% (1.340%, 3.987%), 4.823% (2.630%, 6.559%), 4.216% (1.790%, 5.969%), 1.765% (0.865%, 2.649%), and 0.301% (0.264%, 0.516%), 2.865% (1.455%, 5.768%), 1.033% (0.398%, 1.873%), 1.168% (0.429%, 1.907%), 0.655% (0.283%, 1.652%). The protein expression of Beclin-1 in the tissue of rats where the Freund's complete adjuvant was injected in BCG+ rapamycin group was significantly higher than that of blank control group (Z=4.796, P<0.05). The protein expression of LC3B in the tissue of rats where the Freund's complete adjuvant was injected in BCG alone group was significantly higher than that of blank control group (Z=4.953, P<0.05). Conclusions: Freund's complete adjuvant can enhance the expression levels of local tissue autophagy-related proteins Beclin-1 and LC3B in rat tuberculous wound model.

[Reliability study on establishing a rat tuberculosis wound model using Bacillus Calmette-Guérin].

Objective: To evaluate the reliability of a rat tuberculous wound model established by injecting Bacillus Calmette-Guérin (BCG). Methods: The experimental research was conducted. According to the random number table, fifteen 6-week-old male Sprague-Dawley rats were divided into normal control group and infection group, with 3 rats in normal control group and 12 rats in infection group. Rats in infection group were injected with Freund's complete adjuvant, 3 weeks later, they were injected subcutaneously with BCG bacterial solution to establish a model of tuberculous wounds in rats; rats in normal control group did not receive any treatment. On the 8th, 15th, 32nd, and 43rd day of infection, the skin condition at the injection sites of the rats in infection group was observed roughly. Skin tissue at the injection sites of 3 rats in infection group at each corresponding time point stated above and skin tissue at the corresponding sites of the rats in normal control group were stained with hematoxylin-eosin to observe the cell arrangement, necrosis and inflammation. On 43rd day of infection, acid-fast staining was performed on the skin tissue at the injection sites of the rats in infection group to observe the distribution of bacteria. Results: On the 8th, 15th, 32nd, and 43rd day of infection, tuberculous wound lesions were gradually developed at the skin tissues at the injection sites of the rats in infection group. The cells of the diseased tissue of the rats in infection group arranged disorderly or concentrically, and the number of granulomas and necrotic cells gradually increased, while the skin tissue cells in the corresponding parts of the rats in normal control group arranged regularly with no inflammatory cell infiltration. On the 43rd day of infection, a large number of rod-shaped bacteria were observed in the skin tissue at the injection sites of the rats in infection group. Conclusions: The rat tuberculous wound model established using BCG is stable and reliable, which can meet the experimental requirements.

[Comparative analysis of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis in paraffin-embedded tissue of clinical tuberculous wound].

Objective: To observe the effect of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis (MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods: The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang'an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher's exact probability test, one-way analysis of variance, independent sample t test and Wilcoxon signed rank test. Results: The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups (P=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar (F=1.284, P=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining (t=-12.495, -7.961, P<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue (Z=-3.162, P<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining (t=4.569, 15.519, P<0.01). Conclusions: Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.

Gender-based differences in COVID-19.

The coronavirus disease (COVID-19) is a novel emerging infectious disease spreading worldwide. To further understand the disease, we compared its clinical characteristics, symptoms and outcomes by gender. In an analysis of public surveillance data of Taiwan from January 21 to April 18, 2020, a total of 398 patients were diagnosed with COVID-19 by the detection of severe acute respiratory syndrome coronavirus 2 in pharynx swabs. We divided the patients into two groups: men and women. The associated data were collected, and multivariate comparisons of radiographic infiltration were conducted to analyse the gender-based differences. The mean incubation period was 5.4 ± 5 days, and the incubation period in men was 3.2 days longer than that in women (8 ± 8.1 vs. 4.8 ± 3, p = 0.05). The male patients with COVID-19 with infiltration in chest X-rays (CXR) were 12 years older than their female counterparts. The mortality rate in the male patients with COVID-19 was 6.4-fold higher than that in the female patients (3.2% vs. 0.5%, p < 0.05). The patients with comorbidities of diabetes mellitus and hypertension were vulnerable to infiltration in CXR and the patients with COVID-19 who had infiltration in CXR easily ended up with intubation, intensive care unit admission and mortality. Moreover, female patients with COVID-19 who had fever, cough and dyspnoea were susceptible to infiltration in CXR. Irrespective of whether the cases were imported female from Europe, America or Asia, indigenous male, the factors associated with death in patients with severe COVID-19 were male sex, elderly, female with fever, cough, dyspnoea and DM.

[Research advances in healing mechanism of chronic refractory wounds mediated by long non-coding RNA].

Chronic refractory wound refers to the wound with unclear etiology, multiple and complex injury factors, slow healing, and no obvious tendency of healing after treatment for 4 weeks. The formation and evolution process of chronic refractory wounds are very complex, involving re-epithelialization of wound tissue, cell proliferation, tissue remodeling, and angiogenesis and lymphangiogenesis. The abnormal expression of long non-coding RNA may be involved in the formation of chronic refractory wounds, but the specific pathogenesis and related molecular biological changes are still controversial. In this paper, we reviewed the process and role of long non-coding RNA in regulating keratinocyte differentiation, fibroblast proliferation, and regeneration of vascular and lymphatic endothelial cell in chronic refractory wounds.

Circular RNA ZNF292 affects proliferation and apoptosis of hepatocellular carcinoma cells by regulating Wnt/ß-catenin pathway.

OBJECTIVE: The purpose of this study was to explore the function of circular ribonucleic acid (circRNA) zinc finger protein 292 (ZNF292) in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: The expression of circRNA ZNF292 in Huh-7 cells was knocked down by small interfering RNAs (siRNAs), and the effect of circRNA ZNF292 knockdown on the proliferation of Huh-7 cells was analyzed by Cell Counting Kit-8 (CCK-8) assay and colony formation assay. Then, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) were adopted to analyze the impacts of circRNA ZNF292 knockdown on the cycle distribution and apoptosis of Huh-7 cells. Besides, the influences of circRNA ZNF292 knockdown on Wnt/ß-catenin signaling pathway and its downstream molecules were detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting. RESULTS: Compared with those in siRNA-normal control (NC) group, the proliferation of Huh-7 cells was significantly inhibited and their cloning ability was remarkably weakened (p<0.05), the proportion of cells in S phase was decreased while that in G1 phase was increased (p<0.05), the apoptosis rate of Huh-7 cells was higher and the number of apoptosis was larger in siRNA-2# knockdown group (p<0.05). Besides, in Huh-7 cells with circRNA ZNF292 knockdown, the expressions of Axin, ß-catenin, phosphorylated signal transducer and activator of transcription 3 (p-STAT3), p-STAT5, Cyclin A and Cyclin-dependent kinase 2 (CDK2) were down-regulated, while the expressions of STAT3 and STAT5 did not change remarkably. CONCLUSIONS: Knock downing circRNA ZNF292 leads to cell cycle arrest in G1 phase, thus suppressing cell proliferation and promoting cell apoptosis. The regulatory mechanism of circRNA ZNF292 may involve the regulation of cell cycle and related genes.