Objective: To investigate the urban and rural differences in adverse outcomes of pulmonary tuberculosis (PTB) in patients with pulmonary tuberculosis-diabetes mellitus comorbidity (PTB-DM), so as to provide insights into improving the prevention and treatment measures for PTB-DM. Methods: Patients with PTB-DM who were admitted and discharged from 14 designated tuberculosis hospitals in Hangzhou City from 2018 to 2022 were selected. Basic information, and history of diagnosis and treatment were collected through hospital information systems. The adverse outcomes of PTB were defined as endpoints, and the proportions of adverse outcomes of PTB in urban and rural patients with PTB-DM were analyzed. Factors affecting the adverse outcomes of PTB were identified using a multivariable Cox proportional hazards regression model. Results: A total of 823 patients with PTB-DM were enrolled, including 354 (43.01%) urban and 469 (56.99%) rural patients. There were 112 (13.61%) patients with adverse outcomes of PTB. The proportions of adverse outcomes of PTB in urban and rural patients were 14.41% and 13.01%, respectively, with no statistically significant difference (P>0.05). Multivariable Cox proportional hazards regression analysis identified first diagnosed in county-level hospitals or above (HR=2.107, 95%CI: 1.181-3.758) and drug resistance (HR=3.303, 95%CI: 1.653-6.600) as the risk factors for adverse outcomes of PTB in urban patients with PTB-DM, while the treatment/observed management throughout the process (HR=0.470, 95%CI: 0.274-0.803) and fixed-dose combinations throughout the process (HR=0.331, 95%CI: 0.151-0.729) as the protective factors for adverse outcomes in rural patients with PTB-DM. Conclusions There are differences in influencing factors for adverse outcomes of PTB in urban and rural patients with PTB-DM. The adverse outcomes of PTB are associated with first diagnosed hospitals and drug resistance in urban patients, and are associated with the treatment/observed management and fixed-dose combinations throughout the process in rural patients.

BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.

Objectives:To investigate the value of metagenomic next-generation sequencing (mNGS) in the diagnosis of Pneumocystis jirovecii pneumonia (PJP) in patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) .Methods:The data of 98 patients with suspected pulmonary infection after allo-HSCT who underwent pathogen detection from bronchoalveolar lavage fluid between June 2016 and August 2023 at Nanfang Hospital were analyzed. The diagnostic performance of mNGS, conventional methods, and real-time quantitative polymerase chain reaction (qPCR) for PJP were compared.Results:A total of 12 patients were diagnosed with PJP, including 11 with a proven diagnosis and 1 with a probable diagnosis. Among the patients with a proven diagnosis, 1 was positive by both conventional methods and qPCR, and 10 were positive by qPCR only. Pneumocystis jirovecii was detected by mNGS in all 12 patients. The diagnostic sensitivity of mNGS for PJP was 100%, which was greater than that of conventional methods (8.3%, P=0.001) and similar to that of qPCR (91.6%, P=1.000) . A total of 75% of the patients developed mixed pulmonary infections, and cytomegalovirus and Epstein-Barr virus were the most common pathogens. Mixed infection was detected in eight patients by mNGS and in five patients by qPCR, but not by conventional methods ( P=0.008) . Conclusions:mNGS had good sensitivity for diagnosing PJP after allo-HSCT and was advantageous for detecting mixed infectious pathogens; therefore, mNGS might be an effective supplement to regular detection methods and qPCR.

Objective:To investigate the epidemiology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in patients with maintenance hemodialysis (MHD) in Jiangsu province during SARS-CoV-2 pandemic in China from December 7, 2022 to January 27, 2023, and to analyze the influencing factors of all-cause death.Methods:It was a multi-center cross-sectional investigation. Structured questionnaire was used to collect patient information by medical staff of each hemodialysis center (room) as investigators. Part of the demography data and laboratory examination data came from the Jiangsu Province Hemodialysis Data Information System. MHD patients from hemodialysis centers (rooms) at all levels of medical institutions and independent hemodialysis institutions in Jiangsu province during the outbreak of SARS-CoV-2 infection were included, and the clinical characteristics and all-cause mortality of confirmed and suspected cases of SARS-CoV-2 infection were analyzed.Results:Questionnaire surveys and data analysis on 57 278 patients in 407 hemodialysis centers (rooms) were completed, accounting for 90.41% of the total number of MHD patients (63 357 cases) in Jiangsu province during the same period. There were 24 038 cases (41.97%) of SARS-CoV-2 infection and 14 805 cases (25.85%) of suspected infection, which were widely distributed in all dialysis centers in Jiangsu province. After clinical classification of 38 843 confirmed and suspected SARS-CoV-2 infection cases, 3 662 cases were severe and critical cases, accounting for 9.43% of the infected and suspected cases. Among the patients who had completed the questionnaires, there were 1 812 all-cause deaths, with an all-cause mortality rate of 3.16%. Multivariate logistic regression analysis showed that elderly (taking ≤50 years as a reference, 51-59 years: OR=1.583, 95% CI 1.279-1.933, P=0.001; 60-69 years: OR=3.972, 95% CI 3.271-4.858, P<0.001; 70-79 years: OR=7.236, 95% CI 5.917-8.698, P<0.001; ≥80 years: OR=11.738, 95% CI 9.459-14.663, P<0.001), male ( OR=1.371, 95% CI 1.229-1.529, P<0.001), and co-infection with hepatitis B virus (HBV) (positive serum HBV surface antigen, OR=0.629, 95% CI 0.484-0.817, P<0.001) were independent influencing factors for all cause mortality. Receiver-operating characteristic curve analysis showed that the area under the curve for male, age and current HBV infection prediction of all-cause death was 0.529 ( P<0.001), 0.724 ( P<0.001) and 0.514 ( P=0.042), respectively, and the cut-off value for age prediction of all-cause death was 65.5 years old. Compared with patients without HBV infection, MHD patients with HBV infection significantly reduced the proportion of severe and critically ill patients, all-cause hospitalizations and all cause deaths when infected with SARS-CoV-2 (4.99% vs. 6.41%, χ2=6.136, P=0.013; 8.90% vs. 11.44%, χ2=11.662, P<0.001; 2.01% vs. 3.37%, χ2=10.713, P=0.001, respectively). Conclusion:The MHD patients in Jiangsu province are susceptible to SARS-CoV-2. Elderly age and male gender are independent risk factors for death in MHD patients during the epidemic, while the HBV infection may be a protective factor for death of MHD patients infected with SARS-CoV-2.

The latest epidemiological data suggests that the situation of adult diabetes in China is severe, and metabolic diseases have become significant chronic illnesses that have a serious impact on public health and social development. After more than six years of practice, the National Metabolic Management Center(MMC) has developed distinctive approaches to manage metabolic patients and has achieved a series of positive outcomes, continuously advancing the standardized diagnosis and treatment model. In order to further improve the efficiency, based on the first edition, the second edition guideline was composed by incorporating experience of the past six years in conjunction with the latest international and domestic guidelines.

Dysregulated crosstalk between different signaling pathways contributes to tumor development, including resistance to cancer therapy. In the present study, we found that the mitogen-activated extracellular signal-regulated kinase (MEK) inhibitor trametinib failed to suppress the proliferation of PANC-1 and MGC803 cells by activating the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway, while the JAK2 inhibitor fedratinib failed to inhibit the growth of the PANC-1 cells upon stimulation of extracellular signal-regulated kinase (ERK) signaling. In particular, the most prominent enhancement of the anti-proliferative effect resulted from the concurrent blockage of the JAK2/STAT3 and ERK signaling pathways. Furthermore, the combination of the two inhibitors resulted in a reduced tumor burden in mice. Our evidence suggests novel crosstalk between JAK2/STAT3 and ERK signaling in gastric cancer (GC) and pancreatic ductal adenocarcinoma (PDAC) cells and provides a therapeutic strategy to overcome potential resistance in gastrointestinal cancer.

Objective:To explore the identification value of carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC-Ag), cytokeratin 19 fragment (Cyfra211), thyroglobulin (TG), ferritin (Fer) and procalcitonin (PCT) in fine needle aspiration eluent in benign and malignant cervical nodules, and acquire the optimal diagnostic model.Methods:Three hundred and ninety-six single cervical nodule patients who underwent fine needle aspiration biopsy from August 2017 to August 2019 in the Center Hospital of Xiaogan City of Hubei Province were selected. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG, Fer and PCT were detected by electrogenerated chemiluminescence method. The results of cytopathological diagnosis were regard as "gold standard", and the diagnostic efficiency of single and combined indexes in fine needle aspiration eluent were analyzed by receiver operating characteristic (ROC) curve.Results:Among the 396 patients, malignant nodules was in 101 cases, and benign nodules was in 295 cases. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG and Fer in patients with malignant nodules were significantly higher than those in patients with benign nodules: (27.73 ± 10.63) μg/L vs. (16.81 ± 8.18) μg/L, (1.59 ± 0.74) μg/L vs. (1.09 ± 0.83) μg/L, (3.31 ± 1.48) μg/L vs. (1.66 ± 0.59) μg/L, (144.96 ± 38.93) μg/L vs. (95.03 ± 47.23) μg/L and (191.18 ± 80.13) μg/L vs. (137.87 ± 63.22) μg/L, the PCT was significantly lower than that in patients with benign nodules: (0.61 ± 0.24) μg/L vs. (1.01 ± 0.52) μg/L, and there were statistical differences ( P<0.01). The ROC curve analysis result showed that the CEA, Cyfra211 and TG had super diagnostic value (area under curve>0.7, Youden index>0.5); the area under curve of CEA, Cyfra211 combined with TG was significantly higher than other combined detection of 2 indexes ( P<0.05). Conclusions:The combined detection of CEA, Cyfra211 and TG in fine needle aspiration eluent can effectively distinguish the benign and malignant cervical nodules.

The amino acid sequence of ancestral enzymes from extinct organisms can be deduced through in silico approach termed ancestral sequence reconstruction (ASR). ASR usually has six steps, which are the collection of nucleic acid/amino acid sequences of modern enzymes, multiple sequence alignment, phylogenetic tree construction, computational deduction of ancestral enzyme sequence, gene cloning, and characterization of enzyme properties. This method is widely used to study the adaptation and evolution mechanism of molecules to the changing environmental conditions on planetary time scale. As enzymes play key roles in biocatalysis, this method has become a powerful method for studying the relationship among the sequence, structure, and function of enzymes. Notably, most of the ancestral enzymes show better temperature stability and mutation stability, making them ideal protein scaffolds for further directed evolution. This article summarizes the computer algorithms, applications, and commonly used computer software of ASR, and discusses the potential application in directed evolution of enzymes.
Amino Acid Sequence ; Evolution, Molecular ; Phylogeny ; Proteins/genetics* ; Sequence Alignment

OBJECTIVE：To investigate t he effects of Baihe gujin decoction combined with Shenmai injection on therapeutic efficacy and related indexes in elderly patients with ventilator-associated pneumonia. METHODS ：A total of 75 elderly patients with ventilator-associated pneumonia who were admitted to the Intensive Care Unit （ICU）of Zhuji Hospital of TCM from Apr. 2018 to Oct. 2019 were included. They were randomly divided into control group ，Shenmai group ，Baihe gujin decoction+Shenmai group ， with 25 cases in each group. Control group was given conventional treatments as Piperacillin sodium tazobactam sodium injection 4.5 g，ivgtt，q8 h for anti-infective treatment+ Ambroxol hydrochloride injection 4 mL，iv，bid for expectorant+ Doxofylline injection 20 mL，qd for relieving spasm and asthma+maintaining water ，electrolyte and acid-base balance. Shenmai group was given Shenmai injection 100 mL，ivgtt，qd，on the basis of control group. Baihe gujin decoction+Shenmai group was given Baihe A101） gujin decoction 200 mL nasally ， morning and evening ，on m the basis of Shenmai group. Each group took 7 days as a course of treatment ，and the treatment was continued for 2 courses. Acute physiology and chronic health status （APACHE）Ⅱ score，C-reactive protein （CRP）in peripheral blood ，procalcitonin（PCT），bronchial CD 68 count were observed in 3 groups before and after treatment while the mechanical ventilation time ，hospitalization time in the ICU and ADR recorded. RESULTS：Before treatment ，there was no statistically significant difference in APACHE Ⅱ score，the levels of CRP and PCT ， bronchial CD 68 count among 3 groups（P＞0.05）. After 14 days of treatment ，APACHE Ⅱ score，CRP and PCT levels of all 3 groups were significantly lower than before treatment ；bronchial CD 68 count was significantly higher before treatment ，the order of bronchial CD 68 cell count was Baihe gujin decoction+Shenmai group ＞Shenmai group ＞control group （P＜0.05 or P＜0.01）；after 7 days of treatment ，the order of CRP and PCT levels was Baihe gujin decoction+Shenmai group ＜Shenmai group ＜control group （P＜0.05）；after 14 days of treatment ，there was no staistically significant difference in APACHE Ⅱ score，CRP and PCT levels among 3 groups（P＞0.05）. After treatment ，the order of mechanical ventilation time and ICU hospitalization time was Baihe gujin decoction + Shenmai group ＜Shenmai group ＜control group （P＜0.05 or P＜0.01）. There was no significant difference in the incidence of ADR among 3 groups（P＞0.05）. CONCLUSIONS ：Based on routine treatment ，Baihe gujin decoction combined with Shenmai injection can improve the immune function of lung macrophages in the elderly patients with ventilator-associated pneumonia，inhibite pulmonary inflammation progress ，reduce mechanical ventilation time ，shorten the length of hospital stay with good safety.

Objective To evaluate the effect of serratus plane block (SPB) combined with general anesthesia and SPB with patient-controlled nerve analgesia (PCNA) after surgery on the early postoperative quality of recovery in the patients undergoing radical mastectomy.Methods Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ female patients,aged 18-64 yr,weighing 45-70 kg,scheduled for elective unilateral modified radical mastectomy under general anesthesia,were assigned into 3 groups (n=20 each) using a computer software:SPB and general anesthesia plus patient-controlled intravenous analgesia (PCIA) group (SG+PCIA group),SPB and general anesthesia plus SPB with PCNA group (SG+PCNA group),and general anesthesia plus PCIA group (G+PCIA group).Ultrasound-guided ipsilateral SPB was performed before anesthesia induction.Analgesia was maintained with propofol-remifentanilsevoflurane,rocuronium or vecuronium was intermittently injected to maintain muscle relaxation.PCIA solution contained tramadol 800 mg and flurbiprofen axetil 100 mg in 54 ml of normal saline,and the PCA pump was set up to deliver a 2 ml bolus dose,with a 15 min lockout interval and background infusion at a rate of 0.5 ml/h after a loading dose of 5 ml.The location of the indwelling catheter was confirmed again using ultrasound at the end of surgery,PCNA solution contained 1％ ropivacaine 500 mg in 250 ml of normal saline,and the PCA pump was set up to deliver a 5 ml bolus dose,with a 45 min lockout interval and background infusion at a rate of 5 ml/h after a loading dose of 5 ml.The PCA pumps were used until 48 h after surgery.The automatic key was pressed when pain scores (numerical rating scale [NRS] scores) ≥ 4 at rest or during activity (at 45° ipsilateral upper extremity-up tilt).Quality of Recovery-40 (QoR-40) score was used to assess the early postoperative quality of recovery at 24 and 48 h after surgery.NRS scores at rest or during activity were recorded at 6,8,12,24 and 48 h after surgery.The total pressing times of PCA and occurrence of adverse reactions such as respiratory depression,pruritus,infection at the puncture site or pneumothorax were also recorded.Results Compared with group G+PCIA,the postoperative QoR40 scores were significantly increased,NRS scores at rest or during activity were decreased at each time point after surgery,and the total pressing times of PCA were reduced in SG+PCIA and SG+PCNA groups,the incidence of nausea and vomiting was significantly decreased in group SG+PCIA,and the incidence of nausea and vomiting and dizziness was significantly decreased in group SG+PCNA (P＜0.05 or 0.01).Compared with group SG+PCIA,the postoperative QoR-40 scores were significantly increased at 24 h after surgery,and NRS scores at rest or during activity were decreased after surgery in group SG+PCNA (P＜ 0.05 or 0.01).Conclusion SPB combined with general anesthesia and SPB with PCNA after surgery can raise the early postoperative quality of recovery in the patients undergoing modified radical mastectomy.