Objective:To investigate the efficacy and mechanism of botulinum toxin type A (BTX-A) combined with static progressive stretching (SPS) in the treatment of traumatic knee stiffness in rats.Methods:Forty healthy male SD rats aged 8 weeks and weighing 220-300 g, were randomly divided into blank control group ( n=8) and model groups ( n=28) (including injury group, BTX-A group, SPS group and BTX-A+SPS group, with 7 in each group). Hlidebrand′s method was used to construct a traumatic knee stiffness model, with the following main steps: destruction of the joint capsule, Kirschner wire fixation, joint drilling, and removal of the internal fixation at 4 weeks. The blank control group did not receive any treatment and could move freely in the cage. The injury group moved freely after successful modeling. On the day of internal fixation removal, BTX-A was injected into the joint cavity in group BTX-A, SPS treatment was started in the SPS group, BTX-A was injected into the joint cavity and SPS treatment was started in the BTX-A+SPS group. The treatments lasted 16 days. The range of motion (ROM) and joint stiffness were measured before treatment and at 16 days after treatment. At 16 days after treatment, knee joint tissue was collected and the rats were sacrificed, and the articular capsule fibrous tissue proliferation was observed by HE and Masson staining. The expression levels of phosphorylated (p)-Smad2, Smad2, p-Smad3, Smad3, Smad4, transforming growth factor-β1 (TGF-β1), collagen type I, collagen type III, and α-smooth actin (α-SMA) were determined by Western blot. The ratio of phosphorylated protein to total protein was calculated to reflect the phosphorylation level. Results:(1) ROM: Before treatment, the ROM in the blank control group was significantly higher than that in the other groups ( P<0.05), with no significant difference in ROM among the other groups ( P>0.05). At 16 days after treatment, ROM in the injury group, BTX-A group, SPS group, and BTX-A+SPS group was lower than that in the blank control group ( P<0.05), among which ROM in the BTX-A+SPS group was significantly higher than that in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, there was no significant difference in ROM before and after treatment in the blank control group ( P>0.05), and ROM in the other groups was significantly increased compared with that before treatment ( P<0.01). (2) Joint stiffness: At 16 days after treatment, the joint stiffness levels in the injury group, the BTX-A group, and the SPS group were (0.95±0.24)N·cm/°, (0.86±0.22)N·cm/°, and (0.65±0.09)N·cm/° respectively, which were significantly lower than (0.36±0.03)N·cm/° in the blank control group ( P<0.05). The joint stiffness level of the BTX-A+SPS group was (0.49±0.04)N·cm/°, which was not significantly different from that in the blank control group ( P>0.05), but was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). (3) Fibrous tissue proliferation: at 16 days after treatment, the joint capsular structure in the blank control group was complete and clear, the fibers were arranged in order, and there was no obvious fibrous tissue proliferation. The pathological changes in the injury group were the most serious, with a large number of synovial fibrous tissue proliferation, significantly increased blood vessels in the tissue, and inflammatory cell infiltration. Compared with the SPS group and BTX-A group, the lesions in BTX-A+SPS group were milder, with only slight increase in the number of synovial cells but no obvious vascular proliferation or lymphocytes, and the overall lesions were the least severe. (4) Protein expression: the ratios of p-Smad2/Smad2 in the injury group, BTX-A group and SPS group were 1.552±0.234, 1.328±0.272 and 1.194±0.277 respectively, which were higher than 0.794±0.082 in the blank control group ( P<0.05). The ratio of p-Smad2/Smad2 in the BTX-A+SPS group was 1.013±0.123, which was not significantly different from those in the blank control group, BTX-A group or SPS group ( P>0.05), but was lower than that in the injury group ( P<0.05). At 16 days after treatment, the p-Smad3/Smad3 ratios in the injury group, BTX-A group, SPS group and BTX-A+SPS group were 2.272±0.309, 1.664±0.285, 1.381±0.276 and 1.003±0.060 respectively, which were higher than 0.515±0.051 in the blank control group ( P<0.05). The p-Smad3/Smad3 ratio in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of Smad4 in the injury group (1.001±0.015) was higher than 0.294±0.076 in the blank control group ( P<0.05). However, there was no significant difference between the BTX-A group (0.664±0.051), SPS group (0.833±0.045), BTX-A+SPS group (0.467±0.068) or the blank control group ( P>0.05). The level of Smad4 in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of TGF-β1 in the injury group (1.004±0.407) was higher than 0.269±0.122 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.564±0.194), SPS group (0.422±0.086) and BTX-A+SPS group (0.347±0.161) and the blank control group ( P>0.05). The level of TGF-β1 in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of type I collagen in the injury group was 0.999±0.170, higher than 0.299±0.139 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.542±0.278), SPS group (0.561±0.165), and BTX-A+SPS group (0.537±0.045) and the blank control group ( P>0.05). The level of collagen type I in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, the level of type III collagen in the injury group was 1.002±0.126, higher than 0.239±0.106 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.661±0.062), SPS group (0.595±0.062), and BTX-A+SPS group (0.504±0.269) and the blank control group ( P>0.05). The level of collagen type III in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, the level of α-SMA in the injury group was 0.998±0.074, higher than 0.130±0.023 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.358±0.060), SPS group (0.432±0.230), and BTX-A+SPS group (0.293±0.135) and the blank control group ( P>0.05). The level of α-SMA in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). Conclusions:Compared with single treatment, the combination of BTX-A and SPS demonstrates significantly greater efficacy in the treatment of traumatic knee stiffness in rats. This combined approach not only enhances joint mobility and elasticity but also effectively inhibits joint capsule fibrosis. The underlying mechanism may involve the further suppression of TGF-β1 expression in the joint capsule, leading to reduced phosphorylation levels of Smad2 and Smad3. This, in turn, inhibits the binding of Smad2 and Smad3 to the Smad4 receptor, ultimately downregulating the expression of the downstream proteins of the TGF-β/Smad signaling pathway, such as collagen type I, collagen type III and α-SMA.

Objective:To investigate the efficacy and mechanism of botulinum toxin type A (BTX-A) combined with static progressive stretching (SPS) in the treatment of traumatic knee stiffness in rats.Methods:Forty healthy male SD rats aged 8 weeks and weighing 220-300 g, were randomly divided into blank control group ( n=8) and model groups ( n=28) (including injury group, BTX-A group, SPS group and BTX-A+SPS group, with 7 in each group). Hlidebrand′s method was used to construct a traumatic knee stiffness model, with the following main steps: destruction of the joint capsule, Kirschner wire fixation, joint drilling, and removal of the internal fixation at 4 weeks. The blank control group did not receive any treatment and could move freely in the cage. The injury group moved freely after successful modeling. On the day of internal fixation removal, BTX-A was injected into the joint cavity in group BTX-A, SPS treatment was started in the SPS group, BTX-A was injected into the joint cavity and SPS treatment was started in the BTX-A+SPS group. The treatments lasted 16 days. The range of motion (ROM) and joint stiffness were measured before treatment and at 16 days after treatment. At 16 days after treatment, knee joint tissue was collected and the rats were sacrificed, and the articular capsule fibrous tissue proliferation was observed by HE and Masson staining. The expression levels of phosphorylated (p)-Smad2, Smad2, p-Smad3, Smad3, Smad4, transforming growth factor-β1 (TGF-β1), collagen type I, collagen type III, and α-smooth actin (α-SMA) were determined by Western blot. The ratio of phosphorylated protein to total protein was calculated to reflect the phosphorylation level. Results:(1) ROM: Before treatment, the ROM in the blank control group was significantly higher than that in the other groups ( P<0.05), with no significant difference in ROM among the other groups ( P>0.05). At 16 days after treatment, ROM in the injury group, BTX-A group, SPS group, and BTX-A+SPS group was lower than that in the blank control group ( P<0.05), among which ROM in the BTX-A+SPS group was significantly higher than that in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, there was no significant difference in ROM before and after treatment in the blank control group ( P>0.05), and ROM in the other groups was significantly increased compared with that before treatment ( P<0.01). (2) Joint stiffness: At 16 days after treatment, the joint stiffness levels in the injury group, the BTX-A group, and the SPS group were (0.95±0.24)N·cm/°, (0.86±0.22)N·cm/°, and (0.65±0.09)N·cm/° respectively, which were significantly lower than (0.36±0.03)N·cm/° in the blank control group ( P<0.05). The joint stiffness level of the BTX-A+SPS group was (0.49±0.04)N·cm/°, which was not significantly different from that in the blank control group ( P>0.05), but was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). (3) Fibrous tissue proliferation: at 16 days after treatment, the joint capsular structure in the blank control group was complete and clear, the fibers were arranged in order, and there was no obvious fibrous tissue proliferation. The pathological changes in the injury group were the most serious, with a large number of synovial fibrous tissue proliferation, significantly increased blood vessels in the tissue, and inflammatory cell infiltration. Compared with the SPS group and BTX-A group, the lesions in BTX-A+SPS group were milder, with only slight increase in the number of synovial cells but no obvious vascular proliferation or lymphocytes, and the overall lesions were the least severe. (4) Protein expression: the ratios of p-Smad2/Smad2 in the injury group, BTX-A group and SPS group were 1.552±0.234, 1.328±0.272 and 1.194±0.277 respectively, which were higher than 0.794±0.082 in the blank control group ( P<0.05). The ratio of p-Smad2/Smad2 in the BTX-A+SPS group was 1.013±0.123, which was not significantly different from those in the blank control group, BTX-A group or SPS group ( P>0.05), but was lower than that in the injury group ( P<0.05). At 16 days after treatment, the p-Smad3/Smad3 ratios in the injury group, BTX-A group, SPS group and BTX-A+SPS group were 2.272±0.309, 1.664±0.285, 1.381±0.276 and 1.003±0.060 respectively, which were higher than 0.515±0.051 in the blank control group ( P<0.05). The p-Smad3/Smad3 ratio in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of Smad4 in the injury group (1.001±0.015) was higher than 0.294±0.076 in the blank control group ( P<0.05). However, there was no significant difference between the BTX-A group (0.664±0.051), SPS group (0.833±0.045), BTX-A+SPS group (0.467±0.068) or the blank control group ( P>0.05). The level of Smad4 in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of TGF-β1 in the injury group (1.004±0.407) was higher than 0.269±0.122 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.564±0.194), SPS group (0.422±0.086) and BTX-A+SPS group (0.347±0.161) and the blank control group ( P>0.05). The level of TGF-β1 in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of type I collagen in the injury group was 0.999±0.170, higher than 0.299±0.139 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.542±0.278), SPS group (0.561±0.165), and BTX-A+SPS group (0.537±0.045) and the blank control group ( P>0.05). The level of collagen type I in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, the level of type III collagen in the injury group was 1.002±0.126, higher than 0.239±0.106 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.661±0.062), SPS group (0.595±0.062), and BTX-A+SPS group (0.504±0.269) and the blank control group ( P>0.05). The level of collagen type III in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, the level of α-SMA in the injury group was 0.998±0.074, higher than 0.130±0.023 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.358±0.060), SPS group (0.432±0.230), and BTX-A+SPS group (0.293±0.135) and the blank control group ( P>0.05). The level of α-SMA in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). Conclusions:Compared with single treatment, the combination of BTX-A and SPS demonstrates significantly greater efficacy in the treatment of traumatic knee stiffness in rats. This combined approach not only enhances joint mobility and elasticity but also effectively inhibits joint capsule fibrosis. The underlying mechanism may involve the further suppression of TGF-β1 expression in the joint capsule, leading to reduced phosphorylation levels of Smad2 and Smad3. This, in turn, inhibits the binding of Smad2 and Smad3 to the Smad4 receptor, ultimately downregulating the expression of the downstream proteins of the TGF-β/Smad signaling pathway, such as collagen type I, collagen type III and α-SMA.

Objective:To evaluate the performance, efficacy and safety of a novel portable endoscopy system for upper gastrointestinal examination.Methods:A multicentered, open-label, randomized, non-inferiority controlled study was conducted in 3 clinical research centers from June 2019 to June 2020, and a total of 90 outpatients admitted to Department of Gastroenterology were randomly assigned to the trial group ( n=44) undergoing portable endoscopy and the control group ( n=46) undergoing Olympus endoscopy. The examination success rate, image quality, performance, overall operation satisfaction rate, biopsy success rate and adverse events of the two groups were compared. Results:The examination success rates of the trial group and the control group were 97.73% (43/44) and 100.00% (46/46) respectively with a difference of -2.27% (95% CI: -6.68%-2.13%), higher than the set non-inferiority margin of -10%. Rates of good and excellent image quality were 100% in both groups, and the difference of 0 was higher than the set non-inferiority margin of -10%. There was no significant difference in the rate of good and excellent performance of the operating system between the two groups [97.67% (42/43) VS 100.00% (46/46), P=0.483]. There was significant difference in the overall satisfactory rate of the operation between the two groups [86.05% (37/43) VS 100.00% (46/46), P=0.011]. A total of 9 cases underwent endoscopic biopsy, including 5 cases in the trial group and 4 cases in the control group. The biopsy channels in both groups were smooth and the biopsy were successfully completed. There was no significant difference in adverse event rate between the two groups [25.00% (11/44) VS 10.87%(5/46), χ2=3.07, P=0.080]. All adverse events disappeared in 48 hours, and no severe adverse events or device defect events occurred. Conclusion:The novel portable endoscopic system is comparable to Olympus endoscopic system in terms of the operating performance, the image quality and safety. Therefore, this system is safe and effective for upper gastrointestinal examination.

Objective To establish a quality control system of volatile oil of Chunyang Zhengqi capsules. Methods The chromatogram of volatile oil was established by GC method, and the contents of cinnamaldehyde and eugenol were determined. Results In 15 batches of samples, 19 common peaks were identified, and 9 characteristic peaks were selected to establish the characteristic spectrum. The linear ranges of cinnamaldehyde and eugenol were 0.522 - 1.565 mg/ml (r=0.9994) and 3.038 - 9.115 mg/ml (r=0.9997), respectively. The average recoveries were 97.1% and 97.3%, with RSD of 1.5% and 1.4%, respectively. Conclusion The established GC characteristic map and content determination method could control the quality of essential oil in Chunyang Zhengqi capsules qualitatively and quantitatively. The method is accurate and feasible which could be used as the quality control method of essential oil in Chunyang Zhengqi capsules.

Objective To investigate the application value of magnetic-controlled capsule endoscopy (MCE) for gastric diseases in physical examination of asymptomatic population.Methods Data of 211 asymptomatic individuals who received MCE examinations from July 2015 to December 2016 in Changhai Hospital were collected and rctrospectively analyzed.The tolerance and safety of MCE were studied by analyzing the detection rate for the focal lesions and the rate of endoscopy transfer.Results Among 211 patients,the detection rate of the gastric focal lesions was 9.5％ (20/211).The detection rate in male was higher than that in female (P＜0.05).All patients completed MCE examination successfully and no adverse event was reported.Conclusion MCE,a non-invasive endoscopic modality,is safe and better tolerated than conventional endoscopy,and can be used as a promising approach to screening the gastric diseases in asymptomatic population due to high detection rate of these diseases.

Objective To analyze the clinical characteristics of patients with L isteria septicemia for better clinical diagnosis and management of the disease .Methods A retrospective study was carried out in 13 patients with confirmed diagnosis of Listeria septicemia from July 2009 to November 2013 in West China Hospital of Sichuan University .The clinical features ,laboratory tests ,treatments and clinical outcomes were reviewed and analyzed . Results The vast majority of the 13 patients were immunocompromised or with critical organ dysfunction . The in vitro antimicrobial susceptibility testing indicated that penicillin ,ampicillin and levofloxacin were the most active agents against Listeria ,followed by imipenem ,erythromycin , ciprofloxacin and tetracycline .Only 33 .3% of the 13 Listeria isolates were sensitive to oxacillin .Eight patients were cured ,2 improved ,2 died after therapy .The remaining one patient gave up therapy .Conclusions The incidence of Listeria septicemia was associated with advanced age and presence of underlying diseases .Early etiology diagnosis and appropriate antibacterial therapy can improve the outcome of such patients .Actively treating underlying diseases helps reduce the mortality rate .

Objective To explore the effect of problem-based 1earning (PBL) combined with PICO model in clinical nursing internship teaching.Methods A retrospective case-control study was adopted,133 undergraduate nursing students in grade 2010 were selected as theexperimental group,and 128 undergraduate nursing students in grade 2009 were selected as the control group.The students in the experimental group received a teaching method which combined PBL with PICO model,while the students in the control group received traditional teaching methods.The two groups were evaluated by Objective Structured Clinical Examination (OSCE) and questionnaire survey.Results The OSCE grade and evaluation of teaching method in the experimental group students were higher than the control group,the difference was statistically significant.Conclusions PBL combined with PICO model teaching methods are not only beneficial to cultivate and improve the clinical nursing competency and comprehensive qualities of the nursing students,but also let the students gradually learn and permeate the ideas of any decision made by clinician should be based on the scientific research results,which from the main thought of the evidence-based practice.It can lay a good foundation for going to work in the future,and is also worthy of being popularized and applied in clinical internship teaching of undergraduate nursing students.

Objective To explore the relationship between coagulation/anticoagulation imbalance and oxidative stress in the patients with chronic obstructive pulmonary disease during acute exacerbation (AECOPD)before and after treatment.Methods Plasma tissue factor(TF)and tissue factor pathway inhibitor(TFPl)activity was detected by chromogenic assay in 28 AECOPD patients before and after treatment as well as in 30 healthy controls.The total antioxidative capacity(TAC),malondialdehyde (MDA)and gtutathione peroxidase(GSH-PX)in plasma were measured in both groups.Results The levels of plasma TF and TFPI,and their ratio(TF/TFPI)in AECOPD patients before treatment were significantly higher than those after treatment(all P<0.0 1),the latter were still higher than those in the healthy persons(all P<0.01).The levels of the TAC and GSH-PX in plasma in AECOPD patients before treatment were significantly lower than those after treatment(all P<0.01),the latter were still lower than those in the healthy persons(all P<0.01).The plasma MDA in AECOPD patients before treatment was significantly higher than that after treatment(P<0.0 1),which was still higher than that in the healthy persons(P<0.05).There were negative correlations between TF/TFPI ratio and TAC(r=-0.518.P<0.01),GSH-PX(r=-0.454,P<0.05),PaO2(r=-0.511,P<0.01)respectively and a positive correlation between TF/TFPI ratio and the percentage of neutrophils(r=0.379,P<0.05)in AECOPD patients before treatment.There still were negative correlations between TF/TFPI ratio and TAC (r=-0.420,P<0.05),FEV1% to predicted(r=-0.480,P<0.05)respectively,and a positive correlation between TF/TFPI ratio and MDA(r=0.45 1,P<0.05)in AECOPD patients after treatment.Conclusions There existed coagulation/anticoagulation imbalance and oxidation/antioxidation imbalance before and after treatment in AECOPD patients and their relationship was explored.

Objective　To investigate the HVR-PCR genotype of methicillin-resistant Staphylococci in local hospitals and compare it with the antibiograms, with aview to selecting effective antibacterial agents, moreover, to discuss preliminarily its role in molecular epidemiology. Methods　 The minimal inhibitory concentrations(MICs) of 86 MRSA, 10 MRSE(Mcr S.epidemidis), 5 MSSE(Mcs S.epidemidis), 8 MRSH(Mcr S. haemolyticus) and 5 MSSH(Mcs S. haemolyticus) clinical isolates collected from 4 local hospitals were tested by serial two-fold agar dilution method; their DNA were extracted by moved basic lytic method, whose polymerase chain reaction(PCR) products amplified, based on the size of mec-associated hypervariable region(HVR) were analyzed by PAG vertical and agarose gel electrophoresis. Results 　 MRSA, MRSE and MRSH were grouped into 4, 3 and 2 HVR genotypes respectively according to the size of the PCR products. The PCR products amplified from 9 of 10 MRSE isolates were the same as the products amplified from MRSA isolates. MRSA strains in this study were mainly HVR genotypes A and D, which accounted for 52.32% and 39.53%； Genotypes B and C were the most multi-drug resistant, but genotype D was multi-sensitive. The Ⅰ genotype of MRSE was multi-drug resistant, but its genotype Ⅲ was multi-drug sensitive. The genotype a of MRSH was more resistant than genotype b. Conclusion These results suggest that HVR-PCR genotype method is an easy and fast method for epidemiological investigation of nosocomial infections caused by MRSA， and it is helpful for clinical selection of antibacterial agents. This method can compare the mec determinants of MRSA and Mcr CNSt isolates and hence to search for the origin of the mec determinant.