To screen and identify a chitosanase with high stability, we cloned the chitosanase gene from Bacillus atrophaeus with a high protease yield from the barren saline-alkali soil and expressed this gene in Escherichia coli. The expressed chitosanase of B. atrophaeus (BA-CSN) was purified by nickel-affinity column chromatography. The properties including optimal temperature, optimal pH, substrate specificity, and kinetic parameters of BA-CSN were characterized. The results showed that BA-CSN had the molecular weight of 31.13 kDa, the optimal temperature of 55 ℃, the optimal pH 5.5, and good stability at temperatures below 45 ℃ and pH 4.0-9.0. BA-CSN also had good stability within 4 h of pH 3.0 and 10.0, be activated by K⁺, Na⁺, Mn²⁺, Ca²⁺, Mg²⁺, and Co²⁺, (especially by Mn²⁺), and be inhibited by Fe³⁺, Cu²⁺, and Ag⁺. BA-CSN showcased the highest relative activity in the hydrolysis of colloidal chitosan, and it had good hydrolysis ability for colloidal chitin. Under the optimal catalytic conditions, BA-CSN demonstrated the Michaelis constant K_m and maximum reaction rate V_max of 9.94 mg/mL and 26.624 μmoL/(mL·min), respectively, for colloidal chitosan. In short, BA-CSN has strong tolerance to acids and alkali, possessing broad industrial application prospects.
Bacillus/genetics* ; Hydrogen-Ion Concentration ; Escherichia coli/metabolism* ; Glycoside Hydrolases/biosynthesis* ; Substrate Specificity ; Enzyme Stability ; Chitosan/metabolism* ; Temperature ; Kinetics ; Cloning, Molecular ; Bacterial Proteins/biosynthesis* ; Recombinant Proteins/genetics*

Objective: To establish a progressive research strategy for “colonic components analysis - efficacy verification and mechanism exploration - gut microbiota”, screen pharmacodynamic substances, and investigate their mechanism via gut microbiota. Methods: The pharmacodynamics of Gegen Qinlian decoction (GQD) were assessed using a mouse model of dextran sulfate sodium-induced ulcerative colitis (UC). Ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometer was used to identify the prototype and metabolic components of GQD in the colon during UC. To analyze the structure and function of characteristic genera of GQD and its active components, 16S rRNA sequencing was performed. Results: We identified 67 prototypic and 14 metabolic components of GQD in the UC colon. The primary prototype components are flavonoids and alkaloids, including puerarin (PUE), baicalin (BAI), and berberine (BER). The metabolism was predominantly sulfonation. Efficacy verification showed that the main active components, puerarin, baicalin, and berberine, had good therapeutic effects on UC. The results of 16S rRNA gene sequencing showed that GQD improved UC by regulating the structure and function of the gut microbiota. The abundance of gut microbiota involved in the metabolism of the prototype components was influenced by the corresponding components. The function prediction results showed that PUE was the most comparable to GQD, with 24 consistent pathways. BAI and BER showed comparable gut microbiota regulation pathways. Characteristic pathways of BER include glucometabolic processes. Conclusion This study focused on the key issues in the gut microbiota pathway and developed a progressive research strategy to understand the transformation mechanisms of colonic components. This research systematically analyzed the active components and metabolic transformation of GQD in the colon during the pathological state of UC, as well as changes in the structure and function of the gut microbiota, clarified the mechanism of GQD and its active components in improving UC via the gut microbiota pathway.

Objective:To observe any effect of using a lower limb exoskeleton robot during ankle rehabilitation training on the walking ability of stroke survivors.Methods:Forty-five persons with cerebral apoplexy were randomly divided into a control group, a robot group and a combination group, each of 15. In addition to routine rehabilitation training 5 days a week for 3 weeks, the robot group additionally trained for 10 minutes assisted by a lower limb exoskeleton. The combined group joined that training and additionally undertook 10 minutes of ankle rehabilitation training. Before and after the experiment all of the participants were evaluated using the Fugl-Meyer lower extremity scale (FMA-LE), the Holden functional walking scale (FAC), and for walking speed and step frequency.Results:After treatment, significant improvement was observed in the average FMA-LE score, FAC grade, walking speed and step frequency in all 3 groups. The robot group′s average FMA-LE score, walking speed and step frequency were then significantly better than those of the control group ( P<0.05). Moreover, the average FMA-LE score, step speed and step frequency of the combined group after treatment were (22.67±1.63) min, (0.65±0.05) m/s and (80.80±4.28) steps /min, respectively, significantly better than the other two groups ( P<0.05). Conclusion:Using an exoskeleton robot combined with ankle rehabilitation training can significantly improve the walking of stroke survivors.

Objective: To reveal the molecular mechanism underlying the compatibility of Salvia miltiorrhiza Bge (S. miltiorrhiza, Dan Shen) and C. tinctorius L. (C. tinctorius, Hong Hua) as an herb pair through network pharmacology and subsequent experimental validation. Methods: Network pharmacology was applied to construct an active ingredient-efficacy target-disease protein network to reveal the unique regulation pattern of S. miltiorrhiza and C. tinctorius as herb pair. Molecular docking was used to verify the binding of the components of these herbs and their potential targets. An H9c2 glucose hypoxia model was used to evaluate the efficacy of the components and their synergistic effects, which were evaluated using the combination index. Western blot was performed to detect the protein expression of these targets. Results: Network pharmacology analysis revealed 5 pathways and 8 core targets of S. miltiorrhiza and C. tinctorius in myocardial protection. Five of the core targets were enriched in the hypoxia-inducible factor-1 (HIF-1) signaling pathway. S. miltiorrhiza-C. tinctorius achieved vascular tone mainly by regulating the target genes of the HIF-1 pathway. As an upstream gene of the HIF-1 pathway, STAT3 can be activated by the active ingredients cryptotanshinone (Ctan), salvianolic acid B (Sal. B), and myricetin (Myric). Cell experiments revealed that Myric, Sal. B, and Ctan also exhibited synergistic myocardial protective activity. Molecular docking verified the strong binding of Myric, Sal. B, and Ctan to STAT3. Western blot further showed that the active ingredients synergistically upregulated the protein expression of STAT3. Conclusion The pharmacodynamic transmission analysis revealed that the active ingredients of S. miltiorrhiza and C. tinctorius can synergistically resist ischemia through various targets and pathways. This study provides a methodological reference for interpreting traditional Chinese medicine compatibility.

Objective To comprehensively analyze the effectiveness of exercise training-based respiratory rehabilitation therapy on patients with occupational pneumoconiosis (hereinafter referred to as "pneumoconiosis"). MethodsLiterature on randomized controlled trials of exercise training-based respiratory rehabilitation therapy for pneumoconiosis patients published from the establishment of the database to July 2023 was retrieved from academic systems such as the China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, VIP Database, and China Biology Medicine using bibliometrics method. The RevMan 5.4 software was used for meta-analysis of the selected literatures. Subgroup analysis was conducted to explore the source of study heterogeneity. The funnel plot method was used to test publication bias. Results A total of 55 articles were included, involving 2 436 pneumoconiosis patients in the experimental group and 2 405 pneumoconiosis patients in the control group. The result of random or fixed effect model showed that the six minutes walking distance, the total score of Short from Health Survey-36, forced vital capacity (FVC), forced expiratory volume in one second (FEV₁), FEV₁/FVC, arterial partial pressure of oxygen of pneumoconiosis patients increased after respiratory rehabilitation therapy (all P<0.05), while the total score of the St. George's Respiratory Questionnaire and arterial partial pressure of carbon dioxide decreased compared with the conventional treatment (all P<0.05). The result of subgroup analysis showed that the total score of the St. George's Respiratory Questionnaire, FEV₁, and the index of arterial partial pressure of oxygen of pneumoconiosis patients was better in the rehabilitation treatment for ≥six months compared with those

Objective To compare pregnancy outcomes among pregnant women undergoing cervical polypectomy during different gestational periods.Methods A total of 125 pregnant women were diagnosed with cervical polyps and underwent surgical removal.These participants were divided into the natural pregnancy group(n=71)and the in vitro fertilization and embryo transfer group(IVF-ET group,n=54).Retrospective analysis was conducted on various clinical parameters including age,gestational times,delivery times,abortion times,history of adverse preg-nancy outcomes,history of endometrial polyps,pre-polyp removal vaginal bleeding episodes,gestational age at the time of polyp removal procedure,number of polyps detected during surgery,postoperative pathology results as well as pregnancy outcomes.Results Vaginal bleeding was the most common symptom of pregnancy combined with cer-vical polyps,accounting for 111 cases(88.8％)(62 cases in the natural pregnancy group(87.32％)and 49 ca-ses in the IVF-ET group(90.74％)).Some patients had increased vaginal discharge or were found during a gyne-cological exam,accounting for 11.2％of the patients.All patients underwent cervical polypectomy during pregnan-cy,except for 9 cases of continued pregnancy(4 cases in the natural pregnancy group and 5 cases in the IVF-ET group),of which 13 had miscarriage,10 had preterm birth(6 in the natural pregnancy group and 4 in the IVF group)and 93(54 cases in natural pregnancy group,39 cases in IVF group)(74.4％)delivered at full term.Postoperative pathology was mostly inflammatory polyps(105 cases,84％),deciduous polyps(20 cases,8％)(10 cases in the natural pregnancy group and 10 cases in the IVF-ET group,14.09％and 18.52％,respective-ly).The incidence of preoperative vaginal bleeding,decidual polyp,postoperative abortion,and postoperative pre-term birth in the IVF-ET group exhibited higher rates compared to those observed in the natural pregnancy group;however,these differences did not reach statistical significance.Conclusion The cervical polypectomy procedure was performed during pregnancy for both the natural conception group and the IVF-ET group,with no significant disparity observed in pregnancy outcomes between these two cohorts.

Objective To observe the therapeutic effect of laparoscopic sleeve gastrectomy (LSG) in treating patients with obesity and polycystic ovary syndrome (PCOS). Methods In this retrospective analysis, clinical materials of 60 patients with obesity and PCOS by LSG were analyzed, and postoperative improvements of PCOS symptoms were observed. Results A total of 60 patients had a preoperative mean body mass index (BMI) of (39.4±5.4) kg/m² and an average age of (26.3±6.1) years old, and all of them successfully completed LSG. After follow-up of 12 to 18 months, 50 cases (83.3%) had normalized menstrual cycles, with an average recovery time of (4.5 ±2.2) months after surgery, and 7 cases had natural pregnancy after surgery. Before surgery, ultrasound results in 35 patients indicated polycystic ovarian morphology (PCOM), and 29 cases (82.9%) regained normal ovarian status with an average recovery duration of (6.3±2.8) months. Complicated hyperandrogenemia was observed in 31 patients, and 29 cases (93.5%) exhibited normalization of testosterone levels within 18 months after surgery. At 18 months after surgery, the BMI was (26.47±4.70) kg/m², the average weight loss was (32.4±7.4) %. Conclusion LSG is a safe and effective treatment for obese PCOS.

Objective Investigating the relationship between smoking,overweight,and bibulosity with plasma PAI-1 and osteonecrosis of the femoral head after internal fixation by retrospective study,and providing reference for clinical treatment.Methods A total of 95 patients with femoral neck fracture treated with internal fixation from January 2021 to January 2022 were retrospectively studied.The data of smoking,overweight,alcohol abuse,preoperative and postoperative plasma PAI-1 levels were collected.The patients were divided into necrosis group and non-necrosis group according to the occurrence of femoral head necrosis at the last follow-up.Univariate analysis was performed on smoking,overweight,alcohol abuse,plasma PAI-1 level before and 2 days after operation,and the presence of smoking,overweight and bibulosity factors was analyzed.The association of smoking,overweight and bibulosity with plasma PAI-1 level was analyzed.Figure out the relationship between smoking,overweight,bibulosity and plasma PAI-1 and femoral head necrosis after internal fixation.Results Femoral head necrosis occurred in 22 of 95 patients.Age,gender,smoking,overweight and bibulosity were not independent risk factors for postoperative femoral head necrosis(P＞0.05).The plasma PAI-1 level at 2 days after operation in necrosis group was(46.95±2.35)ng/ml,which was significantly higher than that in non-necrosis group[(41.94±4.73)ng/ml,P＜0.05].The level of plasma PAI-1 at 2 days after operation was(48.29± 1.78)ng/ml in patients with smoking,overweight and alcohol abuse at the same time,which was significantly higher than that in other patients[(42.40±4.60 ng/ml,P＜0.05].In addition,when the smoking,overweight,and bibulosity were concurrent,it was a risk factor for femoral head necrosis(P＜0.05).Conclusion When the smoking,overweight,and bibulosity were concurrent,it can increases the risk of osteonecrosis of the femoral head after internal fixation,which may be related to the significant increase of plasma PAI-1 level,especially at 2 days after surgery.