This paper introduces a real-world cohort research model for community-acquired pneumonia （CAP） based on the Jiangsu Traditional Chinese Medicine （TCM） Dominant Diseases Diagnosis and Treatment Data Platform. Firstly， data cleaning is performed by standardizing diagnosis， symptoms， treatment and imaging， intelligently extracting unstructured information， and cleaning and constructing a standardized database. Secondly， for cohort establishment， CAP patients across the province are screened in accordance with CAP diagnostic criteria to build a high-quality disease-specific cohort. Lastly， in terms of protocol design， the characteristics of TCM research and the CAP disease profile are considered to determine appropriate inclusion and exclusion criteria， estimate sample size， define interventions， outcomes and economic evaluations， providing a reference for real-world TCM research on CAP.

This paper introduces a real-world cohort research model for community-acquired pneumonia （CAP） based on the Jiangsu Traditional Chinese Medicine （TCM） Dominant Diseases Diagnosis and Treatment Data Platform. Firstly， data cleaning is performed by standardizing diagnosis， symptoms， treatment and imaging， intelligently extracting unstructured information， and cleaning and constructing a standardized database. Secondly， for cohort establishment， CAP patients across the province are screened in accordance with CAP diagnostic criteria to build a high-quality disease-specific cohort. Lastly， in terms of protocol design， the characteristics of TCM research and the CAP disease profile are considered to determine appropriate inclusion and exclusion criteria， estimate sample size， define interventions， outcomes and economic evaluations， providing a reference for real-world TCM research on CAP.

Objective To investigate the association between sleep behaviors （including bedtime， sleep quality， and nap duration） and academic performance among college students， as well as the regulatory effect of test anxiety and exercise habit on the association between sleep and academic performance. Methods A questionnaire survey was conducted to collect data. The grade of compulsory courses （with higher grades indicating better academic performance） was used as an ordinal dependent variable. All independent variables were coded by levels： for bedtime during exam and non-exam periods， higher levels indicated later bedtimes； for anxiety during the exam period， higher levels indicated more severe anxiety； for sleep quality during both exam and non-exam periods， higher levels indicated better sleep quality； for nap duration， higher levels indicated longer naps. Exercise habit was treated as a binary variable （yes/no）. Ordinal logistic regression （proportional odds model） was used for multivariable analysis， and OR and 95%CIwere calculated. In addition， sex-stratified models were established to investigate sex differences in the associations between these factors and academic performance. Results A total of 1 221 college students were included， among whom male students accounted for 33.7%. After controlling for confounding factors， bedtime during the exam period was significantly positively associated with the grade of compulsory courses， indicating that later bedtime was associated with a greater likelihood of achieving a higher grade （OR=1.32，95%CI 1.08‒1.62， P=0.007）. Sex was also an independent predictive factor， and the OR of achieving a higher grade in female students was 2.4 times that in male students （OR=2.41， 95%CI 1.90‒3.06， P<0.001）. Sleep quality during the exam period （OR=1.18，95%CI 0.99‒1.42， P=0.070） and exercise habit （OR=1.24， 95%CI 0.99‒1.55， P=0.064） showed marginally significant associations with grade. In contrast， bedtime during the non-exam period， nap duration， anxiety during the exam period， and self-rated sleep quality during the non-exam period showed no statistical significance （all P>0.05）. Sex-stratified analysis showed that among the male students， later bedtime during the exam period was associated with a greater likelihood of achieving a higher grade （OR=1.48， 95%CI 1.05‒2.09， P=0.025）， whereas higher anxiety during the exam period was associated with a lower likelihood （OR=0.79，95%CI 0.63‒0.98， P=0.032）； among the female students， better sleep quality during the exam period was associated with a greater likelihood of achieving a higher grade （OR=1.27， 95%CI 1.02‒1.57， P=0.033）， and bedtime during the exam period showed a positive association with academic performance， but without statistical significance （OR=1.25，95%CI 0.96‒1.62，P=0.091）. Conclusion The association between sleep behaviors and academic performance among college students varies with academic period and sex. Later bedtime during the exam period is associated with better academic performance， but this effect is observed only among male students. In addition， the academic performance of male students is negatively affected by test anxiety， whereas the academic performance of female students is positively associated with subjective sleep quality. Future interventions aimed at improving academic health should consider sex-specific strategies.
Anxiety ; Exercise

Diabetes is a very complex endocrine disease whose common feature is the increase in blood glucose concentration. Persistent hyperglycemia can lead to blindness, kidney and heart disease, neurodegeneration, and many other serious complications that have a significant impact on human health and quality of life. The number of people with diabetes is increasing yearly. The global diabetes prevalence in 20-79 year olds in 2021 was estimated to be 10.5% (536.6 million), and it will rise to 12.2% (783.2 million) in 2045. The main modes of intervention for diabetes include medication, dietary management, and exercise conditioning. Medication is the mainstay of treatment. Marketed diabetes drugs such as metformin and insulin, as well as GLP-1 receptor agonists, are effective in controlling blood sugar levels to some extent, but the preventive and therapeutic effects are still unsatisfactory. Peptide drugs have many advantages such as low toxicity, high target specificity, and good biocompatibility, which opens up new avenues for the treatment of diabetes and other diseases. Currently, insulin and its analogs are by far the main life-saving drugs in clinical diabetes treatment, enabling effective control of blood glucose levels, but the risk of hypoglycemia is relatively high and treatment is limited by the route of delivery. New and oral anti-diabetic drugs have always been a market demand and research hotspot. Inhibitor cystine knot (ICK) peptides are a class of multifunctional cyclic peptides. In structure, they contain three conserved disulfide bonds (C3-C20, C7-C22, and C15-C32) form a compact “knot” structure, which can resist degradation of digestive protease. Recent studies have shown that ICK peptides derived from legume, such as PA1b, Aglycin, Vglycin, Iglycin, Dglycin, and aM₁, exhibit excellent regulatory activities on glucose and lipid metabolism at the cellular and animal levels. Mechanistically, ICK peptides promote glucose utilization by muscle and liver through activation of IR/AKT signaling pathway, which also improves insulin resistance. They can repair the damaged pancrease through activation of PI3K/AKT/Erk signaling pathway, thus lowering blood glucose. The biostability and hypoglycemic efficacy of the ICK peptides meet the requirements for commercialization of oral drugs, and in theory, they can be developed into natural oral anti-diabetes peptide drugs. In this review, the structural properties, activity and mechanism of ICK pattern peptides in regulating glucose and lipid metabolism were summaried, which provided a reference for the development of new oral peptides for diabetes.

Objective:To establish a TaqMan-MGB probe-based method for detecting the polymorphic loci C677T and A1298C of the MTHFR gene.Methods:Specific primers and TaqMan-MGB probes targeting the C677T and A1298C polymorphic loci of the MTHFR gene were designed and optimized based on the gene sequence information.A real-time quantitative PCR detection system was established.Gradient dilution experiments were conducted to determine the limit of detection,and reproducibility experiments were performed to evaluate detection consistency.Specificity was validated using wild-type and mutant plasmid templates.The method was applied to detect 56 clinical samples,and its accuracy and practicality were assessed through comparison with traditional Sanger sequencing.Results:The TaqMan-MGB probe method demonstrated high specificity for detecting the C677T and A1298C loci,with no cross-reactivity between wild-type and mutant probes,enabling accurate genotype differentiation.Sensitivity experiments revealed detection limits of 1.13 × 103 copies/μL for C677T and 8.39 × 101 copies/μL for A1298C.Reproducibility experiments showed coefficients of variation below 1％,indicating stable and reliable results.Among the 56 clinical samples,the overall detection rate for the C677T locus was 86.99％,and for the A1298C locus,it was 97.92％.The TaqMan-MGB method exhibited good concordance with Sanger sequencing results.Conclusion:The TaqMan-MGB method exhibits high specificity,sensitivity,and excellent reproducibility in detecting the polymorphic loci C677T and A1298C of the MTHFR gene,making it suitable for rapid detection in large-scale clinical samples.This method provides an effective molecular diagnostic tool for the early diagnosis and prevention of folate-related diseases.

Objective To investigate the effect of drug-containing serum from Liuwei Dihuang Pill on Wnt/β-catenin pathway,migration and invasion of trophoblast cells induced by high glucose.Methods The drug-containing serum of Liuwei Dihuang Pill was prepared.Human chorionic trophoblast HTR-8/Svneo cells were cultured in vitro and divided into the control group(5 mmol/L glucose+blank serum),the high glucose group(30 mmol/L glucose+blank serum),the low-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.45 g/mL),the medium-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.90 g/mL)and the high-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 1.35 g/mL).The mRNA expressions of Wnt1,β-catenin and Cyclin D1 in HTR-8/Svneo cells were detected by real-time fluorescence quantitative PCR(qRT-PCR).The protein expressions of Wnt1,β-catenin,phosphorylated glycogen synthesis kinase-3β(p-GSK-3β)and Cyclin D1 in HTR-8/Svneo cells were detected by Western blot assay.The migration and invasion of HTR-8/Svneo cells were detected by Transwell method.Results Compared with the control group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells in HTR-8/Svneo cells were decreased in the high glucose group(P<0.05).Compared with the high glucose group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells were significantly increased in the low dose,the medium dose and the high dose groups of the HTR-8/Svneo cells(P<0.05).In addition,with the increase of the drug containing serum dose of Liuwei Dihuang Pill,the above indexes in HTR-8/Svneo cells increased successively(P<0.05).Conclusion The drug containing serum of Liuwei Dihuang Pil can activate Wnt/β-catenin pathway in HTR-8/Svneo cells induced by high glucose,and promote the migration and invasion of HTR-8/Svneo cells.

Objective To analyze the clinical characteristics of children with moyamoya disease and the influence of Chinese medicine identification and acupuncture treatment plan on their prognosis.Methods A retrospective analysis was conducted on the diagnosis and treatment of children diagnosed with moyamoya disease who underwent surgery and were treated with acupuncture at Tianjin Children's Hospital on July 10,2023.The main acupuncture points were as follows:Points on head and face:shangxing,yintang,touwei,middle line of vertex,anterior oblique line of vertex-temporal(the upper 1/5 section),lateral line 1 of vertex,anterior temporal line,renzhong,shanglianquan,sibai,yingxiang,quanliao,xiaguan,and dicang;Points on upper extremities:jiquan,chize,neiguan(quick stabbing without leaving needles),jianyu,jianliao,quchi,waiguan,yangxi,houxi,hegu;Points on lower extremities:weizhong,weiyang(quick stabbing without leaving needles),biguan,xuehai,liangqiu,zusanli,qiuxu,sanyinjiao,and taichong),the needles were performed once a day,and continuous treatment for 5 days each week,and the general conditions of the children were observed after 2 weeks of treatment,and the Fugl-Meyer assessment(FMA),the Barthel index(BI),and the simple test for evaluating hand function(STEF)were used,to assess left upper limb motor function;neurophysiologic techniques were used to examine motor nerve conduction in the left limb.Results The child was an 8-year-old male admitted to the hospital on July 10,2023.①Complaint:unfavorable movement of the left limb for more than 1 month.②History:the child had an abnormal seizure during waking hours 2 years before admission,which lasted for about 1 minute and then resolved on its own,and he could recall the course of the seizure afterward.Attended at the department of neurology of our hospital and diagnosed with moyamoya disease after being discharged from the hospital,the child was found at home to have weakness of the left limbs,unable to lift the upper limbs and unable to walk on the lower limbs.Attended at the department of neurosurgery of a certain hospital and was diagnosed with acute cerebral infarction and underwent cranio-cerebral vascular anastomosis.After the operation,the child's left limb activity is unfavorable,the right limb has no motor disorder and sensory abnormality,speech expression is unclear,to further improve the motor function of the child's limbs,the patient was referred to the rehabilitation department of our hospital,the initial diagnosis of moyamoya disease disease recovery(left hemiplegia),according to the specific conditions of the adjustment of the acupuncture program,2 weeks after the treatment,the patient's facial lines were basically symmetrical,crying and laughing,the corner of the mouth does not droop.After 4 weeks of treatment,the child was able to walk on the ground independently,but hemiplegic gait,left lower limb muscle tone basically relieved,proximal muscle strength grade Ⅲ-Ⅳ,distal muscle strength grade Ⅱ-Ⅲ.After 6 weeks of treatment,the child's hemiplegic gait was improved,proximal muscle strength was grade Ⅳ,distal muscle strength was grade Ⅲ-Ⅳ,the left hand could hold objects for a few minutes,left thumb flexor strength was grade Ⅲ,dorsal extensor strength was grade Ⅱ,and the rest of the four fingers flexor and dorsal extensor strength was grade Ⅳ.The proximal muscle strength was grade Ⅴ and distal muscle strength was grade Ⅳ.The left wrist could move freely,the left interphalangeal joint could extend and hold objects,and the motor function of the left upper limb was basically established,and the condition was stabilized.FMA,BI and STEF scores all increased with the prolongation of the treatment time,and the latency and nerve conduction velocity of the median nerve,peroneal nerve and tibial nerve tended to normal after treatment.Conclusion The efficacy of traditional acupuncture in treating pediatric moyamoya disease after cerebral infarction surgery rehabilitation is remarkable,which can significantly improve the clinical symptoms of children recovering from moyamoya disease.

Objective:To analyze the related factors of biochemical recurrence after laparoscopic radical prostatectomy (LRP).Methods:Clinical data of 312 patients with prostate cancer who received LRP in Hunan Provincial People's Hospital (the First Affiliated Hospital of Hunan Normal University) from September 2020 to September 2023 were retrospectively analyzed. Those patients were followed up for 1 year after surgery. According to whether biochemical recurrence occurred after surgery, the above patients were divided into biochemical recurrence group ( n=61) and non-biochemical recurrence group ( n=251). Compare the clinical data [including age, smoking history, drinking history, history of diabetes, postoperative Gleason score, body mass index (BMI), tumor stage, presence or absence of seminal vesicle invasion, and positive surgical margins] and laboratory indicators [including fasting blood glucose after admission, hemoglobin, prostate volume, prostate mass, and preoperative prostate-specific antigen (PSA)] levels between the two groups of patients. Measurement data with normal distribution were presented as Mean±SD, comparison between the two groups was performed by the independent samples t-test. Categorical data were expressed as case (%), comparison between groups was performed by the χ2 test, multivariate analysis was performed by Logistic regression model. Results:The difference in smoking history between groups was not statistically significant( P>0.05). The proportions of patients with age≥65 years old [85.25% (52/61) vs. 68.13% (171/251)], alcohol drinking history[73.77% (45/61) vs. 68.53% (172/251)], diabetes mellitus[50.82% (31/61) vs. 40.24% (101/251)], Gleason score >7 points[72.13% (44/61) vs. 30.68% (77/251)], BMI≥30 kg/m 2[32.79% (20/61) vs. 14.34% (36/251)], disease stage ≥T2a[67.21% (41/61) vs. 23.11% (58/251)], seminal vesicle invasion[32.79% (20/61) vs. 13.55% (34/251)], and postoperative positive incisal margin[24.59% (15/61) vs. 3.59% (9/251)] in the biochemical recurrence group were higher than those in the non-biochemical recurrence group, and the differences were statistically significant ( χ2=393.22, 34.58, 74.28, 36.70, 725.49, 48.09, 171.37, 30.49, respectively, all P<0.001). The differences in fasting blood glucose [(6.81±0.89) mmol/L vs. (6.63±0.78) mmol/L], prostate volume[37.19±4.23) mL vs. (36.87±4.36) mL], prostate weight[(48.21±5.11) g vs. (47.82±5.13) g], and hemoglobin level[(134.08±15.62) g/L vs. (133.26±16.24) g/L] between both groups of patients were not statistically significant ( t=1.57,0.52,0.53,0.36, P=0.117,0.606,0.594,0.722, respectively). The preoperative prostate-specific antigen (PSA) level was higher in the biochemical recurrence group than that in the non-biochemical recurrence group [(45.13±5.26) μg/L vs. (28.87±3.18) μg/L, t=30.99, P<0.001). Multivariate Logistic regression analysis results indicated that postoperative Gleason score >7 points, preoperative PSA and seminal vesicle invasion were risk factors for postoperative biochemical recurrence in patients undergoing LRP ( OR=5.39, 95% CI:1.57-18.48, P=0.008; OR=4.32, 95% CI:1.32-14.10, P=0.016; OR=12.76, 95% CI:1.47-111.03, P=0.022). Conclusion:Patients with prostate cancer are prone to biochemical recurrence after LRP, which is affected by postoperative Gleason score, preoperative PSA level and seminal vesicle invasion. It is necessary to take targeted measures to prevent biochemical recurrence.

Objective:To establish a TaqMan-MGB probe-based method for detecting the polymorphic loci C677T and A1298C of the MTHFR gene.Methods:Specific primers and TaqMan-MGB probes targeting the C677T and A1298C polymorphic loci of the MTHFR gene were designed and optimized based on the gene sequence information.A real-time quantitative PCR detection system was established.Gradient dilution experiments were conducted to determine the limit of detection,and reproducibility experiments were performed to evaluate detection consistency.Specificity was validated using wild-type and mutant plasmid templates.The method was applied to detect 56 clinical samples,and its accuracy and practicality were assessed through comparison with traditional Sanger sequencing.Results:The TaqMan-MGB probe method demonstrated high specificity for detecting the C677T and A1298C loci,with no cross-reactivity between wild-type and mutant probes,enabling accurate genotype differentiation.Sensitivity experiments revealed detection limits of 1.13 × 103 copies/μL for C677T and 8.39 × 101 copies/μL for A1298C.Reproducibility experiments showed coefficients of variation below 1％,indicating stable and reliable results.Among the 56 clinical samples,the overall detection rate for the C677T locus was 86.99％,and for the A1298C locus,it was 97.92％.The TaqMan-MGB method exhibited good concordance with Sanger sequencing results.Conclusion:The TaqMan-MGB method exhibits high specificity,sensitivity,and excellent reproducibility in detecting the polymorphic loci C677T and A1298C of the MTHFR gene,making it suitable for rapid detection in large-scale clinical samples.This method provides an effective molecular diagnostic tool for the early diagnosis and prevention of folate-related diseases.

Objective To investigate the effect of drug-containing serum from Liuwei Dihuang Pill on Wnt/β-catenin pathway,migration and invasion of trophoblast cells induced by high glucose.Methods The drug-containing serum of Liuwei Dihuang Pill was prepared.Human chorionic trophoblast HTR-8/Svneo cells were cultured in vitro and divided into the control group(5 mmol/L glucose+blank serum),the high glucose group(30 mmol/L glucose+blank serum),the low-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.45 g/mL),the medium-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.90 g/mL)and the high-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 1.35 g/mL).The mRNA expressions of Wnt1,β-catenin and Cyclin D1 in HTR-8/Svneo cells were detected by real-time fluorescence quantitative PCR(qRT-PCR).The protein expressions of Wnt1,β-catenin,phosphorylated glycogen synthesis kinase-3β(p-GSK-3β)and Cyclin D1 in HTR-8/Svneo cells were detected by Western blot assay.The migration and invasion of HTR-8/Svneo cells were detected by Transwell method.Results Compared with the control group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells in HTR-8/Svneo cells were decreased in the high glucose group(P<0.05).Compared with the high glucose group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells were significantly increased in the low dose,the medium dose and the high dose groups of the HTR-8/Svneo cells(P<0.05).In addition,with the increase of the drug containing serum dose of Liuwei Dihuang Pill,the above indexes in HTR-8/Svneo cells increased successively(P<0.05).Conclusion The drug containing serum of Liuwei Dihuang Pil can activate Wnt/β-catenin pathway in HTR-8/Svneo cells induced by high glucose,and promote the migration and invasion of HTR-8/Svneo cells.