1.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
2.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
3.Study on the impact of long-term blood glucose fluctuations and blood glucose control on the occurrence of stroke in patients with type 2 diabetes mellitus
Zhihao HU ; Shuai ZHU ; Yuhua LI ; Anle LI ; Fang XIANG ; Hongjie YU ; Yiying ZHANG
Chinese Journal of Diabetes 2024;32(2):101-107
Objective Explore the impact of long-term blood glucose fluctuations and blood glucose control on the stroke incidence in patients with type 2 diabetes mellitus(T2DM).Methods An observa-tional cohort of patients with T2DM was established based on"Shanghai Community Chronic Disease Health Management Service Objects"on October 1st,2018.Follow-ups were conducted every three months,and fasting blood glucose(FBG)were tested at each visit.Basic epidemiological data were collected via the Shanghai Community Health Management Information Platform or survey questionnaires,and stroke incidents were gathered via the"Shanghai Cardio-Cerebrovascular Event Monitoring System".The first reported stroke incident within the observation period was considered.The observational deadline was December 31st,2021.Standard deviation of FBG was used to evaluate blood glucose fluctuation and FBG control rate was used to reflect blood glucose control status.Cox Proportional Hazards Model was utilized to analyze the impacts.Results The cumulative observation time was 91,826.1 person-years for the study,in which there were 1785 cases of stroke events observed,implying a cumulative incidence of 5.73%and incidence rate of 1943.9/100000 person-years.The mean number of follow-ups(10.29±3.07)per patient was recorded.The details of the stroke cases were as follows:transient ischemic attacks(n=111,6.22%),non-lacunar brain infarctions(n=754,42.24%),lacunar brain infarctions(n=798,44.71%),intracerebral hemorrhages(n=80,4.48%),non-intracerebral hemorrhages(n=8,0.45%),subarachnoid hemorrhages(n=12,0.67%),and unclassified strokes(n=22,1.23%).After excluding subjects with less than five follow-ups,the Cox Proportional Hazards Model suggested that increased standard deviation of FBG was an independent risk factor for the first stroke incident,and an increased blood glucose control rate was an independent protective factor.Both the standard deviation of FBG and the blood glucose control rate were not correlated with stroke recurrence.Conclusion Long-term blood glucose fluctuations and blood glucose control can predict the risk of a first stroke in patients with T2DM.The relationship between blood glucose fluctuations,blood glucose management,and stroke recurrence requires further studies.
4.Exploration of Therapeutic Effect of Wujiwan on Inflammatory Bowel Disease in Rats Based on PPARγ Signaling Pathway and T-cell Immunoregulation
Shiyun GUO ; Yuxuan GUO ; Yi SUN ; Xiaoxin ZHU ; Yujie LI ; Ying CHEN ; Qing YANG ; Yajie WANG ; Qi LI ; Xiaogang WENG ; Zhihao DENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):237-245
ObjectiveThis study explores the efficacy and pharmacological mechanism of Wujiwan in rats with inflammatory bowel disease (IBD) from the perspectives of the peroxisome proliferator-activated receptor γ (PPARγ) signaling pathway and T-cell immunity, providing reference for the treatment of IBD with traditional Chinese medicine. MethodThe study involved administering 2,4,6-trinitrobenzenesulfonic acid (TNBS) enemas to 35 rats to induce acute IBD. After 24 hours, the animals were divided into the following groups: normal group, model group, Wujiwan treatment group, and positive drug control group. Each group received gastric gavage for 8 consecutive days before the rats were dissected to compare the disease activity index (DAI) of the rat colon tissue, the colon mucosal damage index (CMDI), and the spleen index. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interleukin-1β (IL-1β), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α) in the serum. Quantitative real-time polymerase chain reaction (Real-time PCR) was used to determine the mRNA expression levels of T-bet (T-box expressed in T cells) and Gata3 (Gata-binding protein-3) in the colon tissue. Western blot analysis was conducted to detect the protein expression levels of PPARγ, T-bet, and nuclear factor-κB p65 (NF-κB p65) in the rat colon. ResultThe rat model of IBD was successfully established. Compared with the model group, the Wujiwan treatment group showed reduced DAI, CMDI, and spleen index, decreased content of TNF-α in the serum(P<0.01), significantly increased content of IL-10(P<0.01), and elevated mRNA content of T-bet and Gata3(P<0.05) in the colon tissue. The expression of PPARγ protein was augmented(P<0.05), and the expression of T-bet and NF-κB p65 protein was decreased(P<0.05,P<0.01). ConclusionWujiwan activates or upregulates PPARγ expression in IBD rats to inhibit the generation of pro-inflammatory factors, participates in the inflammatory immune process, and alleviates inflammatory reactions. Its mechanism may involve regulating the NF-κB pathway through PPARγ, enhancing Th2 cell transcription expression, and reducing Th1 cell transcription.
5.A Review of Theoretical Research on Interpretation of Scientific Connotation of Compatibility of Traditional Chinese Medicine Compounds
Shiyun GUO ; Zhihao DENG ; Yan LI ; Yuxuan GUO ; Xiaoxin ZHU ; Yujie LI ; Ying CHEN ; Qing YANG ; Yi SUN ; Yajie WANG ; Qi LI ; Weiyan CAI ; Xiaogang WENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):338-344
Traditional Chinese medicine (TCM) has a long history of application in China and has consistently played a vital role in treating diseases and saving lives. TCM prescriptions (compounds) constitute the primary form of clinical TCM treatment and significantly differ from western medicine (chemicals) due to the diverse composition and chemical constituents of TCM (compounds). Nevertheless, the potential multi-component, multi-target, and multi-pathway action characteristics of TCM prescriptions also demonstrate their possible (complementary) therapeutic advantages when compared with single-component chemical drugs. Therefore, driven by the development of modern science and technology and the demands of the modernization and internationalization of TCM, modern theories regarding the complexity of TCM prescription effects have been continuously proposed: Different from the abstract language of traditional prescription theory, the modern TCM prescription theory is more inclined to illustrate the connotation of prescription compatibility concretely and vividly from an experimental and scientific perspective. In this paper, new theories on the complexity of TCM prescriptions proposed in recent years are summarized to provide research references and ideas for the greater role of TCM prescriptions and a better scientific understanding.
6.Construction of digital intelligent minimally invasive research-oriented hepatobiliary and pancreatic surgery discipline
Jian YANG ; Xiwen WU ; Wen ZHU ; Haoyu HU ; Nan XIANG ; Ning ZENG ; Zhihao LIU ; Xuequan FANG ; Chihua FANG
Chinese Journal of Digestive Surgery 2024;23(1):91-97
The emergence and evolution of digital intelligent technology has profoundly influenced the development of minimally invasive research-oriented hepatobiliary and pancreatic surgery discipline. Over various periods, our team has always adhered to the principle of "being oriented by clinical issues and driven by clinical needs", continuously carried out innovative research across interdisciplinary boundaries, propelling the evolution of digital intelligent technology. Spanning over two decades, this journey includes the progression from digital virtual human, three-dimensional visualization, molecular fluorescence imaging, augmented reality and mixed reality, artificial intelligence, to the realm of human visualization meta-universe. This evolution facilitates the shift from two-dimensional empirical diagnoses of hepatobiliary and pancreatic surgical diseases to deep learning intelligent diagnostics, the transition from morphology-based tumor diagnoses to molecular imaging-based diagnostics, and from conventional empirical surgery to intelligent navigation surgery. The authors provide a comprehensive review of our developmental process and achievements within the realm of digital intelligent diagnostic and therapeutic technologies, with the aims to promote the development and application of digital intelligent medicine.
7.Establishment of a qPCR method to detect Staphylococcus xylosus and its application
Lingzhi YU ; Liping FENG ; Zhihao KONG ; Qi ZHU ; Xiaofeng WEI
Acta Laboratorium Animalis Scientia Sinica 2024;32(1):73-79
Objective To establish and evaluate a method for rapid and sensitive S.xylosus detection using qPCR(real-time quantitative PCR).Methods A gehM gene fragment was selected as the target for S.xylosus.A set of specific primers was synthesized and a qPCR method was established to detect S.xylosus.A S.xylosus standard strain and other non-target strains were chosen for analysis.DNA of S.xylosus was diluted 10-fold to determine its sensitivity.Clinical samples were tested,and positive products were sequenced.The result were compared with those of bacterial culture.Results S.xylosus had a specific amplification curve,whereas other non-S.xylosus species did not,indicating that the primers were specific for S.xylosus.Sensitivity was 100 fg/μL DNA.Repeatability within and between groups was less than 3%.A total of 60 clinical samples were analyzed,of which five samples had a typical S curve.qPCR products were sequenced and BLAST searched.The similarity of the gene sequences was 99.63%,indicating that the sample was positive for the S.xylosus gehM gene with a positivity rate of 8.3%.However,the positivity rate of bacterial culture was 6.7%.The positivity rate of qPCR was slightly higher than that of the culture.Conclusions The established qPCR method is rapid with high sensitivity and specificity,and can be used to detect S.xylosus.
8.Current Situation,Existing Problems and Solving Strategies of the Assessment of Medical Ethics and Styles in the Examination of Doctors' Qualification
Hua GUO ; Yu HAN ; Kai REN ; Zhihao ZHOU ; Mingjie ZI ; Yujiao SONG ; Min ZHU ; Lili XYU ; Yi LI ; Tingting WANG ; Linyong YU
Journal of Traditional Chinese Medicine 2024;65(12):1292-1295
This paper discussed the current education status on medical ethics and styles and the assessment condition in the examination of doctors' qualification, as well as the existing problems and potential solutions by reviewing domestic and foreign literature and summarizing the practice experience. Traditionally, medical ethics and styles have always been integrated into clinical medical practice in China. However, under the modern medical education system, it is challenged to integrate traditional education on medical ethics and styles with the rules of modern medical knowledge. By summarizing the education and assessment status of medical ethics and styles in the examination of doctors' qualification, it is found that the current examination is relatively poor in the evaluation content, and the way of evaluation is not diverse, with lack of curriculum of medical humanities. The solutions suggested are enriching relevant examination content, introducing more and comprehensive evaluation method, and establishing more medical humanities-related courses.
9.Multicenter retrospect analysis of early clinical features and analysis of risk factors on prognosis of elderly patients with severe burns
Qimin MA ; Wenbin TANG ; Xiaojian LI ; Fei CHANG ; Xi YIN ; Zhaohong CHEN ; Guohua WU ; Chengde XIA ; Xiaoliang LI ; Deyun WANG ; Zhigang CHU ; Yi ZHANG ; Lei WANG ; Choulang WU ; Yalin TONG ; Pei CUI ; Guanghua GUO ; Zhihao ZHU ; Shengyu HUANG ; Liu CHANG ; Rui LIU ; Yongji LIU ; Yusong WANG ; Xiaobin LIU ; Tuo SHEN ; Feng ZHU
Chinese Journal of Burns 2024;40(3):249-257
Objective:To investigate the early clinical characteristics of elderly patients with severe burns and the risk factors on prognosis.Methods:This study was a retrospective case series study. Clinical data of 124 elderly patients with severe burns who met the inclusion criteria and were admitted to the 12 hospitals from January 2015 to December 2020 were collected, including 4 patients from the Fourth People's Hospital of Dalian, 5 patients from Fujian Medical University Union Hospital, 22 patients from Guangzhou Red Cross Hospital of Jinan University, 5 patients from Heilongjiang Provincial Hospital, 27 patients from the First Affiliated Hospital of Naval Medical University, 9 patients from the First Affiliated Hospital of Nanchang University, 10 patients from Affiliated Hospital of Nantong University, 9 patients from Tongren Hospital of Wuhan University & Wuhan Third Hospital, 12 patients from the 924 th Hospital of PLA, 6 patients from Zhangjiagang First People's Hospital, 4 patients from Taizhou Hospital of Zhejiang Province, and 11 patients from Zhengzhou First People's Hospital. The patients' overall clinical characteristics, such as gender, age, body mass index, total burn area, full-thickness burn area, inhalation injury, causative factors, whether combined with underlying medical diseases, and admission time after injury were recorded. According to the survival outcome within 28 days after injury, the patients were divided into survival group (89 cases) and death group (35 cases). The following data of patients were compared between the two groups, including the basic data and injuries (the same as the overall clinical characteristics ahead); the coagulation indexes within the first 24 hours of injury such as prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time, D-dimer, fibrinogen degradation product (FDP), international normalized ratio (INR), and fibrinogen; the blood routine indexes within the first 24 hours of injury such as white blood cell count, platelet count, neutrophil-to-lymphocyte ratio, monocyte count, red blood cell count, hemoglobin, and hematocrit; the organ function indexes within the first 24 hours of injury such as direct bilirubin, total bilirubin, urea, serum creatinine, aspartate aminotransferase, alanine aminotransferase, total protein, albumin, globulin, blood glucose, triglyceride, total cholesterol, alkaline phosphatase, creatine kinase, electrolyte indexes (potassium, sodium, chlorine, calcium, magnesium, and phosphorus in blood), uric acid, myoglobin, and brain natriuretic peptide; the infection and blood gas indexes within the first 24 hours of injury such as procalcitonin, C-reactive protein, pH value, oxygenation index, base excess, and lactate; treatment such as whether conducted with mechanical ventilation, whether conducted with continuous renal replacement therapy, whether conducted with anticoagulation therapy, whether applied with vasoactive drugs, and fluid resuscitation. The analysis was conducted to screen the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns. Results:Among 124 patients, there were 82 males and 42 females, aged 60-97 years, with body mass index of 23.44 (21.09, 25.95) kg/m 2, total burn area of 54.00% (42.00%, 75.00%) total body surface area (TBSA), and full-thickness burn area of 25.00% (10.00%, 40.00%) TBSA. The patients were mainly combined with moderate to severe inhalation injury and caused by flame burns. There were 43 cases with underlying medical diseases. The majority of patients were admitted to the hospital within 8 hours after injury. There were statistically significant differences between patients in the 2 groups in terms of age, total burn area, full-thickness burn area, and inhalation injury, and PT, APTT, D-dimer, FDP, INR, white blood cell count, platelet count, urea, serum creatinine, blood glucose, blood sodium, uric acid, myoglobin, and urine volume within the first 24 hours of injury (with Z values of 2.37, 5.49, 5.26, 5.97, 2.18, 1.95, 2.68, 2.68, 2.51, 2.82, 2.14, 3.40, 5.31, 3.41, 2.35, 3.81, 2.16, and -3.82, respectively, P<0.05); there were statistically significant differences between two groups of patients in whether conducted with mechanical ventilation and whether applied with vasoactive drugs (with χ2 values of 9.44 and 28.50, respectively, P<0.05). Age, total burn area, full-thickness burn area, serum creatinine within the first 24 hours of injury, and APTT within the first 24 hours of injury were the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns (with odds ratios of 1.17, 1.10, 1.10, 1.09, and 1.27, 95% confidence intervals of 1.03-1.40, 1.04-1.21, 1.05-1.19, 1.05-1.17, and 1.07-1.69, respectively, P<0.05). Conclusions:The elderly patients with severe burns had the injuries mainly from flame burns, often accompanied by moderate to severe inhalation injury and enhanced inflammatory response, elevated blood glucose levels, activated fibrinolysis, and impaired organ function in the early stage, which are associated with their prognosis. Age, total burn area, full-thickness burn area, and serum creatinine and APTT within the first 24 hours of injury are the independent risk factors for death within 28 days after injury in this population.
10.Feasibility and safety of stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath
Haijie XIE ; Junkai HUANG ; Zhihao FU ; Fu ZHU ; Linguo XIE ; Chunyu LIU
Chinese Journal of Urology 2024;45(8):614-618
Objective:To investigate the feasibility and safety of a treatment system for stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath.Methods:The clinical data of 72 patients with upper urinary calculi admitted to the Second Hospital of Tianjin Medical University from November 2022 to February 2023 were retrospectively analyzed. All patients achieved stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath. There were 50 males and 22 females. The average age was (54.7±12.1) years. Preoperative urine culture was positive in 14 cases, negative in 3 cases (4.2%)with nitrite positive, and 11 cases were negative for urine culture and nitrites but positive of white blood cells (+ + + ). There were 29 cases of renal calculi, 33 cases of upper ureteral calculi, and 10 cases of upper ureteral calculi combined with renal calculi.The mean stone diameter was 17.0(14.0, 24.0)mm. CT value was (1 049.3±258.6)HU. Twenty-four patients carried ureteral stents before operation. A total of 42 cases used ureteral sheaths with diameters of F11/13, and 30 cases used sheaths with diameters of F12/14.During the operation, an infusion pump was used to provide sufficient irrigation pressure. The negative pressure suction was attached to the distal end of the sheath. The flexible head of the sheath was guided to the target renal calyx, to completely aspirate stone fragments. Stone baskets was used in 11 cases during the procedure. The level of ureteral injury was assessed according to the Traxer grading system at the end of the operation. A ureteral stent with extraction string was retained.On the first day postoperatively, CT scanning was performed to evaluate the residual stone fragments. Patients were discharged on postoperative day 2-3 after the removal of the ureteral stent and catheter. Follow-up was conducted for 30 days postoperatively, during which the Ureteral Stent Symptom Questionnaire (USSQ) was used to assess voiding symptoms and pain. Painkiller usage and emergency revisit situations were recorded. CT scans were performed to evaluate the stone-free rate on postoperative day 30.Results:The average operation time was 30.0 (20.0, 44.5) minutes. A total of 70 cases had no ureteral injuries, and 2 cases had Grade 1 ureteral injuries (minor mucosal damage). Three cases developed fever within 72 hours postoperatively, with no cases of septic shock or fever after stent removal. Eight patients reported waist and abdominal discomfort after discharge and took oral pain medication. Among them, one patient returned to the emergency department for pain treatment. Five patients reported moderate or severe genitourinary symptoms (including voiding frequency, nocturia, urgency/incontinence, dysuria, hematuria, and incomplete emptying) based on subjective evaluation. All patients could work and recovered a normal daily life after discharge and there was no readmission or additional surgical procedures. There were 61 patients achieved immediate stone-free status on the first day after surgery, and 66 patients achieved stone-free status during follow-up at 30 days postoperatively.Conclusions:Stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath is safe and feasible.

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