The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

Abstract Esophageal cancer is one of the most common malignant tumors of the digestive system in China , with e-sophageal squamous cell carcinoma (ESCC) being the predominant pathological type. The exact etiology of ESCC remains incompletely understood. With advances in 16S rRNA gene sequencing and metagenomics , microbial dys- biosis has been suggested to play a significant role in the pathogenesis of ESCC. Currently , research on the rela- tionship between microorganisms and ESCC is still in its early stages. This review summarizes the association be- tween oral and esophageal microbiota and ESCC , factors influencing microbial composition , and microbial commu- nities linked to ESCC prognosis , which may contribute to early detection and optimized treatment strategies for ES- CC.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

Objective:To investigate the value of BRAF V600E and multigene detection and stratified application for the diagnosis of thyroid nodules.Methods:A total of 1 117 patients with thyroid nodules resection at Nanjing Gulou Hospital from December 2020 to July 2022 were enrolled in the study. Fine needle aspiration (FNA) and core biopsy samplings were performed for cytopathologic examination and genetic testings; the findings were combined with BSRTC classification. The diagnostic performance of BRAF V600E and multigene detection were compared.Results:Among the 1, 117 patients who underwent thyroid nodules resection, 285 were male and 832 were female, with a median age of 46 years (range: 24-76 years). Postoperative histopathologic examination confirmed 1 040 cases of thyroid cancer and 77 cases of benign nodules. The sensitivity (87.0% vs. 80.8%, P<0.01) and diagnostic accuracy (87.9% vs. 82.1%, P<0.01) of multigene detection were significantly higher than those of BRAF V600E detection. The result of multigene detection showed that BRAF V600E mutation was the most common finding, followed by CCDC6-RET (E1-E12) fusion, ETV6-NTRK3 fusion, and KRAS mutation. Multigene detection had a higher sensitivity (81.9% vs. 72.8%, P<0.01) and lower cancer risk in wild-type (47.6% vs. 57.7%, P=0.069) than BRAF V600E detection in BSRTCⅠ-Ⅴ lesions. Compared with BRAF V600E detection, multigene had no significant difference of sensitivity in BSRTC Ⅰ lesions, but significantly higher sensitivity (86.3% vs 74.0%, P<0.01) in BSRTC Ⅲ lesions. Conclusions:Genetic detection can be used as an effective tool for the diagnosis of thyroid nodules. A stratified application of molecular markers in the diagnosis of thyroid nodules is proposed. Combined with FNA, single gene or multigene detection both can effectively assist in the diagnosis of thyroid nodules. Moreover, multigene detection is superior to single gene detection. For BSRTC Ⅲ lesion with wild-type BRAF, multigene detection can be considered with a repeated FNA.

Objective To investigate the effect of resistance training on the rehabilitation of patients after hip replacement with the framework of the theory of planned behavior(TPB).Methods A total of 85 patients of hip fracture patients in Gansu Provincial Hospital of Traditional Chinese Medicine from July 2022 to April 2023 were selected as research objects.The patients were separated into intervention group(n=43)and control group(n=42)by random sampling.Both groups were treated with routine nursing,and the intervention group was given resistance training based on TPB.The intervention time was 3 months.The Harris hip score(HHS),36-item short form(SF-36)score and Barthel index(BI)scores of the two groups were compared before intervention,1 month and 3 months after intervention,as well as the functional exercise compliance score of the patients 3 months after intervention.Results After 1 month and 3 months of intervention,the HHS,SF-36 score and BI score of intervention group were better than those of control group,and after 3 months of intervention,the executive ability of functional exercise in intervention group was significantly higher than that of control group,and the mean difference was significant(P＜0.05).Conclusion Resistance training led by TPB can effectively improve the functional exercise compliance of patients after total hip arthroplasty,improve the muscle strength,balance ability and joint stability of patients,and then improve the daily activity ability and quality of life.

BackgroundChronic obstructive pulmonary disease （COPD） is a common chronic respiratory disease， and patients with COPD often experience substantially emotional difficulties， such as anxiety and depression， all of which may cause serious detriment to the prognosis of patients. As a non-pharmacological intervention in clinical practice， group mindfulness-based stress reduction therapy （MBSR） is beginning to emerge， while has rarely been studied in COPD patients with concurrent emotional difficulties. ObjectiveTo evaluate the effects of group MBSR on depression， state of mindfulness and pulmonary function in stable COPD patients， so as to provide references for the application of group MBSR in patients with COPD. MethodsA total of 97 patients with stable COPD who were followed up in the Department of Respiratory and Critical Care Medicine of Mianyang Third People's Hospital from January to October 2019 were selected as the study objects， and they were assigned into study group （n=50） and control group （n=47） by random number table method. All individuals received routine medication therapy and an 8-week health education， based on this， participants in study group partook an 8-week intervention comprising group MBSR. At the baseline， 4 weeks and 8 weeks of intervention， participants were assessed with Self-rating Depression Scale （SDS）， Five Facet Mindfulness Questionnaire （FFMQ） and COPD Assessment Test （CAT）， as well as the pulmonary function testing. ResultsThere were 41 patients in study group and 42 cases in control group completed the study. The group * time interaction was interpreted as significant between two groups for SDS， FFMQ and CAT scores （F=54.858， 86.161， 69.862， P<0.01）. Baseline SDS， FFMQ and CAT scores of the two groups yielded no statistical difference between two groups （F=0.240， 0.052， 0.019， P>0.05）， while study group scored lower on SDS and CAT （F=12.900， 38.511， 7.797， 28.824， P<0.01） and higher on FFMQ （F=27.324， 82.412， P<0.01） than those of the control group after 4 and 8 weeks of intervention. With the prolongation of intervention time in study group， participants demonstrated an overall reduction in SDS and CAT scores （F=109.753， 124.144， P<0.01）， and an increase in FFMQ scores （F=228.194， P<0.01）. There were no between-group differences in forced expiratory volume in one second as percentage of predicted volume （FEV1%pred） after 4 and 8 weeks of intervention （F=0.104， P=0.748） ， and the within-group changes in FEV1%pred value over the intervention period in study group was not statistical （F=0.561， P=0.458）. ConclusionGroup MBSR may help relieve depressive symptoms， enhance mindfulness level， and alleviate clinical symptoms in stable COPD patients， but has no effect on pulmonary function. ［Funded by Mianyang Health and Health Commission Scientific Research Project （number， 201916）］

Objective: To investigate the relationship between hypertriglyceridemic waist (HTW) phenotype and abnormal blood pressure in children and adolescents in Yinchuan City, Ningxia, and to provide scientific basis for early identification and prevention of hypertension. Methods: A cluster sampling method was adopted to select 1 566 children and adolescents in Yinchuan. All participants completed questionnaire survey, physical measurement and biochemical examination. Results: In general, after adjusting for age, smoking, drinking, and family history of hypertension, compared with the normal triglyceride normal waist (NWNT) phenotype, the enlarged waist (EW) and HTW phenotypes both increased the risk of abnormal blood pressure ( OR EW =2.62, 95% CI =1.87-3.67; OR HTW =3.97, 95% CI =2.30-6.86); the risk of abnormal blood pressure in boys with EW and HTW phenotypes was that of boys with NTNW phenotype, respectively 3.80 times (95% CI =2.35-6.15) and 3.32 times (95% CI =1.59-6.92) of girls EW and HTW phenotypes. The risk of abnormal blood pressure in girls with EW and HTW phenotypes was that of girls with NTNW phenotype, respectively 1.72 times (95% CI =1.06-2.82) and 4.62 times(95% CI =1.97-10.85). Conclusion The hypertriglyceridemic waist phenotypes among children and adolescents in Yinchuan is significantly correlated with increased risk of abnormal blood pressure. More attention should be paid to triglyceride level and waist circumference among children and adolescents.

Objective: To explore the relationship between the muscle mass index (MMI) and the aggregation of cardiovascular risk factors (CVRFs) in children and adolescents, and to provide a scientific basis for the prevention and treatment of cardiovascular diseases in children and adolescents. Methods: The current situation study design was adopted, and 1 622 children and adolescents aged 12-17 years old in Yinchuan City were selected by a cluster random sampling method. All subjects were subjected to questionnaire survey, physical examination, body composition determination and laboratory index testing. Results: When other variables were not adjusted, MMI was a risk factor for the aggregation of cardiovascular risk factors ( P <0.01). After adjusting for age, gender and BMI, MMI became a protective factor for CVRFs≥1 ( OR =0.74, 95% CI =0.62-0.89), compared with insufficient MMI, the risk of developing CVRFs≥1 with good MMI and sufficient MMI was 0.60(95% CI =0.46-0.79), 0.56(95% CI =0.37- 0.85 ) times. The risk of CVRFs≥2 was 0.54(95% CI =0.37-0.79), 0.51(95% CI =0.30-0.87) times, and similar results were found in boys ( P <0.05). Conclusion Under the same BMI level, muscle mass index is a protective factor for cardiovascular risk factor aggregation in children and adolescents. Physical exercise of children and adolescents should be emphasized to maintain the best muscle mass and weight.

Objective: To analyze the relationship between body fat distribution and bone mineral content (BMC) in adolescents and gender differences among Chinese adolescents, and to provide a scientific basis for the prevention and treatment of bone metabolic diseases. Methods: A total of 1 575 adolescents aged 12-18 years old were selected from Yinchuan by cluster random sampling. Body composition and bone mineral content were measured by bioelectrical impedance analysis(BIA). Multiple linear regression analysis was used to explore the relationship between body fat distribution and BMC after adjusting for confounding factors. Results: Except for LTFR, the levels of FM, AFM, TFM, TrTFR and TrLFR in boys were lower than those in girls( t =-13.52， -15.18 ，-12.47，-12.25，-7.96， P <0.05); After adjusting for age, sex and weight, FM, AFM, TFM, TrTFR and TrLFR were all negatively correlated with BMC level( P <0.05). For each increase of 1 standard deviation in TFM, BMC level decreased by 0.53 standard deviation(95% CI=-0.57--0.49, P<0.01]. LTFR had a linear negative correlation with BMC level in boys( B=0.07, P <0.01), no similar correlation was found in girls( B=0.01, P =0.74). There was a linear negative correlation between TrLFR and BMC level in girls( B=-0.06, P =0.03), but the correlation was of no significance in boys( B=-0.01, P =0.55). Conclusion Sex difference in body fat distribution in Chinese adolescents is observed. Body fat distribution is closely related to bone minerals content in adolescents.

Objective: To investigate the mechanism of moxibustion in the treatment of diarrhea-predominant irritable bowel syndrome (IBS-D), by observing the effects of moxibustion at Tianshu (ST25) and Shangjuxu (ST37) on microRNA-133b (miRNA-133b), pituitary homeobox family factor 3 (Pitx3)/tyrosine hydroxylase (TH), and neurotransmitters in the brain tissue of IBS-D rats. Methods: Healthy Sprague-Dawley rats were randomly divided into a normal group, a model group, a moxibustion group, and a Western medicine group, with 12 rats in each group. Except for the normal group, the IBS-D rat model was established by mother-offspring separation and acetic acid enema combined with restraint stress stimulation in all the other groups. No intervention was performed in the normal and model groups. Mild moxibustion was applied to both Tianshu (ST25) and Shangjuxu (ST37) in the moxibustion group. Rifaximin was given by gavage in the Western medicine group. The physical status of rats in each group was observed at different periods. After the intervention, hematoxylin- eosin staining was performed to observe the histopathological morphology of rat colon; enzyme-linked immunosorbent assay was used to measure the levels of dopamine (DA), noradrenaline (NE), and 5-hydroxytryptamine (5-HT) in plasma, colon, and midbrain tissue of rats; the relative expression levels of miRNA-133b, Pitx3 mRNA, and TH mRNA in the midbrain tissue were measured by real-time fluorescence quantitative polymerase chain reaction, and the relative expression levels of Pitx3 and TH proteins in the midbrain tissue were measured by Western blotting and immunofluorescence. Results: The body weights of rats among groups and at different time points were statistically different (P<0.01). The body weight of the normal group was higher than that of the other groups over time (P<0.01). After modeling, the minimum volume threshold of abdominal withdrawal reflex (AWR) was significantly lower (P<0.01) and the loose stool rate was significantly higher (P<0.01) in the model, moxibustion, and Western medicine groups compared with the normal group; the miRNA-133b expression in the midbrain tissue was significantly lower (P<0.01), the expression levels of Pitx3 and TH in the midbrain tissue were significantly higher (P<0.01), and the levels of DA, NE, and 5-HT in plasma, colon and midbrain tissue were significantly higher (P<0.01). After the intervention, the minimum volume threshold of AWR was significantly higher (P<0.01), the loose stool rate was significantly lower (P<0.01), the miRNA-133b expression was significantly increased (P<0.01 or P<0.05) and the expression levels of Pitx3 and TH were significantly decreased (P<0.01) in the midbrain tissue, the levels of DA, NE, and 5-HT in plasma, colon, and midbrain tissue were significantly reduced (P<0.01) in the moxibustion and Western medicine groups compared with the model group; the levels of 5-HT in the colon and midbrain tissue of the moxibustion group were significantly lower than those in the Western medicine group (P<0.05), and there was no statistical difference compared with the remaining groups (P>0.05). Linear correlation analysis showed that miRNA-133b was negatively correlated with Pitx3 (r<0, P<0.01); Pitx3 with TH, TH with DA, and NE with 5-HT were positively correlated (r>0, P<0.01).Conclusion: Moxibustion at Tianshu (ST25) and Shangjuxu (ST37) improves diarrhea symptoms and visceral hypersensitivity in IBS-D rats. The mechanism may be related to up-regulating miRNA-133b, inhibiting Pitx3/TH, and reducing neurotransmitter expression levels in the midbrain tissue.