Objective: To investigate whether Tuina alleviates fibrotic symptoms in myofascial trigger points (MTrPs) by regulating transforming growth factor (TGF)-β1/Smad3 signaling pathway, thereby deactivating these points. Methods: This study comprised two experimental phases. In phase 1, 27 specific pathogen-free (SPF) grade female Sprague-Dawley (SD) rats were randomized into three groups: control 1, model 1, and Tuina 1 groups. Model 1 and Tuina 1 groups underwent an 8-week MTrPs modeling protocol involving blunt impact and eccentric exercise. After successful modeling, rats in Tuina 1 group received manual pressing on nodules or cord-like taut bands on the medial aspect of the left hindlimb. Pain sensitivity and tissue stiffness were evaluated via pressure pain threshold (PPT) and soft tissue tension (STT). Muscle histopathology and fibrosis were observed using hematoxylin and eosin (HE) and Masson staining. Inflammatory factors in muscle were measured by enzyme-linked immunosorbent assay (ELISA), while immunofluorescence (IF) and Western blot (WB) were used to detect the expression levels of α-smooth muscle actin (α-SMA), collagen Ⅲ, and TGF-β1. In phase 2, 45 SPF female SD rats were randomized into five groups: control 2, model 2, Tuina 2, TGF-β1 inhibitor (TI), and Tuina + TGF-β1 agonist (Tuina + TA) groups. All groups except control 2 underwent standardized MTrPs modeling. Rats in Tuina 2 group received consistent pressing manipulation. TI group received intraperitoneal injections of oxymatrine, while Tuina + TA group received intraperitoneal injections of SRI-011381 hydrochloride followed by the same pressing protocol as Tuina 2 group. WB was used to detect the expression of collagen I, collagen III, TGF-β1, and phosphorylated-Smad3 (p-Smad3)/Smad3. Results: In phase 1, Tuina significantly improved PPT and STT in MTrPs of rats (P < 0.01), reversed pathological damages including disorganized muscle fiber arrangement, abnormal myocyte morphology, and exacerbated fibrosis. In addition, in MTrPs of rats in model 1 group, expression levels of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and fibrosis markers (α-SMA, collagen I, and collagen III) were upregulated, and all exhibited a significant downward trend after Tuina intervention (P < 0.05 or P < 0.01). This indicates that the therapeutic effects of Tuina are directly associated with reduced local inflammation and fibrosis in MTrPs. In phase 2, compared with model 2 group, rats in TI and Tuina 2 groups had decreased expression levels of TGF-β1 and p-Smad3/Smad3 in MTrPs, alongside reduced levels of inflammatory factors (IL-1β, IL-6, NF-κB, and TNF-α) and fibrosis markers (α-SMA, collagen I, and collagen III) (P < 0.05 or P < 0.01). When co-administered with TGF-β1 agonist, the therapeutic effects of Tuina were significantly attenuated, with rebounded TGF-β1 expression and p-Smad3/Smad3 in local MTrPs, and fibrosis and inflammatory responses were re-exacerbated (P < 0.05 or P < 0.01). Conclusion Tuina can effectively reduce inflammatory responses and fibrosis in MTrPs tissue, and its mechanism is closely related to the inhibition of the TGF-β1/Smad3 signaling pathway, which plays a critical role in Tuina-mediated regulation of MTrPs fibrosis.

Persistent left superior vena cava is a rare venous variant that is often combined with cardiovascular malformations. In thoracic surgery, especially mediastinal tumor resection, neglect of this variant may make the surgery difficult and risky, and careful preoperative imaging interpretation and adequate preoperative evaluation play an important role in the perioperative safety of the patient. In this paper, we reported a case of a 17-year-old female patient with a persistent left superior vena cava combined with mediastinal tumors. She was successfully discharged 5 days after thoracoscopic surgery, and after 3 years of postoperative follow-up, no tumor recurrence was observed.

Objective To establish a gas chromatography-mass spectrometry (GC-MS) method for the simultaneous determination of four carbonate compounds (CCs), including ethyl methyl carbonate (EMC), diethyl carbonate (DEC), vinylene carbonate (VC), and ethylene carbonate (EC) in workplace air. Methods Vapor-phase EMC, DEC, VC, and EC in workplace air were collected using activated carbon tubes. After desorption with dichloromethane, the samples were analyzed by GC-MS. Qualitative identification was performed based on retention times and characteristic ions, while quantitative analysis was conducted using peak areas of selected characteristic ions. Results The quantitative determination ranges for the four CCs were from 0.57×10⁻³ to 200.00 mg/L, with correlation coefficients ≥0.999 45. The detection limit ranged from 0.17 to 1.60 μg/L, and the lower limit of quantification ranged from 0.57 to 5.33 μg/L. The minimum detection concentration and minimum quantitation concentration were 0.11-1.07 and 0.38-3.55 μg/m³, respectively. Mean spiked recoveries ranged from 85.70% to 111.65%. The intra- and inter-batch relative standard deviations were 0.11%-2.04% and 1.27%-5.18%, respectively. Mean desorption efficiencies of the method ranged from 74.70% to 118.20%. EMC, DEC, and EC samples were stable for up to five days at 4 °C, while VC samples were stable for up to three days at 4 °C. Conclusion The GC-MS method is suitable for the simultaneous determination of the four CCs including EMC, DEC, VC, and EC in workplace air.

Objective:To investigate the current status of endoscopic treatment for gastroesophageal varices in portal hypertension in China, and to provide supporting data and reference for the development of endoscopic treatment.Methods:In this study, initiated by the Liver Health Consortium in China (CHESS), a questionnaire was designed and distributed online to investigate the basic condition of endoscopic treatment for gastroesophageal varices in portal hypertension in 2022 in China. Questions included annual number and indication of endoscopic procedures, adherence to guideline for preventing esophagogastric variceal bleeding (EGVB), management and timing of emergent EGVB, management of gastric and isolated varices, and improvement of endoscopic treatment. Proportions of hospitals concerning therapeutic choices to all participant hospitals were calculated. Guideline adherence between secondary and tertiary hospitals were compared by using Chi-square test.Results:A total of 836 hospitals from 31 provinces (anotomous regions and municipalities) participated in the survey. According to the survey, the control of acute EGVB (49.3%, 412/836) and the prevention of recurrent bleeding (38.3%, 320/836) were major indications of endoscopic treatment. For primary [non-selective β-blocker (NSBB) or endoscopic therapies] and secondary prophylaxis (NSBB and endoscopic therapies) of EGVB, adherence to domestic guideline was 72.5% (606/836) and 39.2% (328/836), respectively. There were significant differences in the adherence between secondary and tertiary hospitals in primary prophylaxis of EGVB [71.0% (495/697) VS 79.9% (111/139), χ2=4.11, P=0.033] and secondary prophylaxis of EGVB [41.6% (290/697) VS 27.3% (38/139), χ2=9.31, P=0.002]. A total of 78.2% (654/836) hospitals preferred endoscopic therapies treating acute EGVB, and endoscopic therapy was more likely to be the first choice for treating acute EGVB in tertiary hospitals (82.6%, 576/697) than secondary hospitals [56.1% (78/139), χ2=46.33, P<0.001]. The optimal timing was usually within 12 hours (48.5%, 317/654) and 12-24 hours (36.9%, 241/654) after the bleeding. Regarding the management of gastroesophageal varices type 2 and isolated gastric varices type 1, most hospitals used cyanoacrylate injection in combination with sclerotherapy [48.2% (403/836) and 29.9% (250/836), respectively], but substantial proportions of hospitals preferred clip-assisted therapies [12.4% (104/836) and 26.4% (221/836), respectively]. Improving the skills of endoscopic doctors (84.2%, 704/836), and enhancing the precision of pre-procedure evaluation and quality of multidisciplinary team (78.9%, 660/836) were considered urgent needs in the development of endoscopic treatment. Conclusion:A variety of endoscopic treatments for gastroesophageal varices in portal hypertension are implemented nationwide. Participant hospitals are active to perform emergent endoscopy for acute EGVB, but are inadequate in following recommendations regarding primary and secondary prophylaxis of EGVB. Moreover, the selection of endoscopic procedures for gastric varices differs greatly among hospitals.

BACKGROUND:Pressing massage applied to myofascial trigger points(MTrPs)has shown clear effect in relieving pain.However,further research is needed to investigate the effects of different levels of pressure applied during the massage. OBJECTIVE:To investigate the different strength of pressing on MTrPs in rats with chronic pain. METHODS:Sixty SPF-rated male Sprague-Dawley rats were randomly divided into a blank group of 10 rats not involved in MTrPs modeling and 50 rats involved in modeling.The MTrPs model was established in the left medial thigh muscle of rats by blunt strikes combined with centrifugal exercise and 40 rats that met the evaluation criteria after modeling were randomly divided into model group,light press group,medium press group and heavy press group,with 10 rats in each group.The rats in the blank group and the model group were not intervened,while the rats in the light press group,the medium press group and the heavy press group were intervened with a homemade press stimulator with light force(0.3 kg),medium force(0.5 kg)and heavy force(0.7 kg)to MTrPs.The intervention time was 7.5 minutes per session,with one session every other day,totaling seven sessions.Electromyogram,soft tissue tension and mechanical pain threshold were detected by electrophysiological instruments,soft tissue tension tester,and pressure painmeter,respectively.After the intervention,in the blank group,muscle tissue was taken from the inside of the left thigh,while in the other groups,MTrPs tissue was taken.The pathological morphology was observed by hematoxylin-eosin staining,while enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of cyclooxygenase-2,prostaglandin E2 and bradykinin. RESULTS AND CONCLUSION:Compared with the blank group,the model group had lower mechanical pain thresholds,higher soft tissue tone,higher amplitude frequency of spontaneous electrical activity,significant pathomorphological changes,and increased levels of cyclooxygenase-2,prostaglandin E2 and bradykinin(P<0.05).Compared with the model group,the medium press group and the heavy press group showed increased mechanical pain thresholds,decreased soft tissue tension,decreased spontaneous potential frequency amplitude,and decreased levels of cyclooxygenase-2,prostaglandin E2 and bradykinin(P<0.05),and significant recovery on pathomorphological changes.No significant changes in the above indicators were observed in the light press group(P>0.05).Compared with the medium press group,the heavy press group showed better improvement in the above indicators(P<0.05).To conclude,moderate to heavy pressing is often required to alleviate MTrPs pain.

Objective:To establish collection methods and laboratory testing methods for qualitative and quantitative analysis of 9 typical active pharmaceutical ingredient in the workplace air.Methods:In December 2021, a mixed solution of nine analytes was prepared and then dispersed in aerosol state to simulate sampling. Glass fiber filter membrane was selected as air collector and collected active pharmaceutical ingredient in the air at a rate of 2.0 L/min for 15 minutes. Then, the obtained filter membrane samples were eluted with 25%ACN/75%MeOH. Finally, the eluent was qualitatively and quantitatively analyzed with liquid chromatography-triple quadrupole mass spectrometer.Results:This method could effectively collect active pharmaceutical ingredient in the air, with an average sampling efficiency of more than 98.5%. The linear correlation coefficient r was greater than 0.9990. The lower limit of quantification for each analyte ranged from 0.6~500.0 ng/ml, and the average recovery rate ranged from 97.6%~102.5%.Conclusion:This method could simultaneously collect 9 active pharmaceutical ingredient in the workplace air, and could provide accurate qualitative and quantitative analysis in subsequent laboratory tests.

Objective To analyze the serum PD-L1 levels of gastric cancer patients and its relationship with tumor stage and prognosis.Methods Serum of sixty patients with pathological diagnosis of gastric cancer in our hospital from August 2018 to December 2019 were collected,and the serum of 15 healthy adults as a control.The serum PD-L1 levels was measured by enzyme linked immunosorbent assay(ELISA).The serum PD-L1 levels between gastric cancer patients and healthy adults were compared.The relationship between the serum PD-L1 levels and prognosis of gastric cancer patients with different stages was analyzed.Finally,the PD-L1 expression in tumor tissues was detected by immunohistochemical staining,and its correlation with serum PD-L1 levels was analyzed.Results The serum PD-L1 levels of gastric cancer group was significantly higher than that of the control group.The serum PD-L1 levels in patients with stage Ⅲ and Ⅳ gastric cancer was significantly higher than that in stage Ⅰ and Ⅱ.The serum PD-L1 levels in gastric cancer patients with peritoneal metastasis was significantly elevated,indicating poor prognosis.Kaplan-Meier survival analysis showed that the survival time of gastric cancer patients with higher level of serum PD-L1 was shorter than those with lower ones.We further evaluated the PD-L1 expression in tumor tissues.There was no significant correlation between the serum PD-L1 levels and that in tumor tissues.Conclusions The serum PD-L1 levels of gastric cancer patients was higher than that of healthy adults,and which gradually increased with the progression of the disease.Gastric cancer patients with peritoneal metastasis had a much higher serum PD-L1 levels,and there was no significant correlation between serum PD-L1 levels and PD-L1 expression in tumor tissues.

Objective:To investigate the current status of the technological pedagogical content knowledge (TPACK) levels of youth teachers in a military medical university, and to provide a practical basis for improving youth teachers' ability to teach with technologies.Methods:Youth teachers (with less than 3 years of teaching experience) in a military medical university were selected by convenience sampling for a survey on TPACK abilities using a modified TPACK questionnaire. The questionnaire was distributed through the internet, and 119 valid responses were returned. SPSS 20.0 software was used to perform the independent samples t-test and one-way analysis of variance.Results:The youth teachers scored highest in technological knowledge [TK; (3.68±0.74)] and lowest in TPACK (3.17±0.74). There were no significant differences in TPACK abilities between youth teachers in different majors. Youth teachers of different educational attainments showed significant differences in the levels of technological knowledge (TK; F=12.99, P<0.001), content knowledge (CK; F=13.64, P<0.001), technological pedagogical knowledge (TPK; F=4.38, P=0.015), technological content knowledge (TCK; F=3.83, P=0.025), and TPACK ( F=3.63, P=0.029)—the TK, CK, TPK, and TCK levels of teachers with doctoral degrees were significantly higher than those with bachelor's and master's degrees. Conclusions:Efforts should be made to design a TPACK ability development system throughout the growth cycle of youth teachers, build teaching practice communities, and improve the system guarantees.

In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

In order to establish a rapid,specific and sensitive detection method for Streptococcus equi subspecies zooepidemicus(SEZ)and to understand the infection status of SEZ in horses ente-ring China,specific primers were designed and synthesized based on the conserved gene comB of standard strain SEZ(ATCC 43079)in this work.Then,the pMD19-T-comB recombinant plasmid was constructed and used as a standard positive template.After that,the fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green Ⅰ dye was established.Totally,477 equine entry serum samples from 6 countries,including Netherlands,Belgium,Japan,Germa-ny,Argentina and New Zealand,during 2018 to 2023,were randomly selected and detected for SEZ by the qPCR method.Results showed that the established qPCR method had specific amplification for only SEZ,which illustrated a good specificity.Sensitivity test of the method showed that the limited detection amount was 4.58 X101 copies/μL.And the repeatability test showed that the coef-ficient of variation of intra-batch repeatability was less than 0.5％,while the inter-batch repeat-ability was less than 3.0％,which indicated good repeatability and high stability.Retrospective a-nalysis showed that totally 11 of 477 positive samples were detected,with a relatively low positive rate of 2.31％(11/477).Among them,all the 40 samples from Netherlands in 2018 were negative(0/40).In the samples of 2019,one positive was detected from Belgium(1/20),while all other 36 samples which form Japan and Germany were negative.In the samples of 2021,three samples(3/34)from Japan and one sample(1/20)from Argentina were positive,and all the other 40 samples from the Netherlands were negative.In the samples of 2022,76 samples from Netherlands were all negative.While in the 2023,5(5/126)of 126 samples from Netherlands and one(1/88)of 88 from New Zealand were found positive with SEZ.To summarize,The SYBR Green Ⅰ qPCR method for the diagnosis of SEZ was successfully established,and it could provide necessary technical support for the rapid quarantine of China's entry-exit and port departments,as well as the epidemiological investigation of the disease.