Objective To investigate the efficiency and mechanism of C1q tumor necrosis factor-related protein 3(CTRP3)on reprogramming of cardiac fibroblasts(CFs)into induced cardiomyocyte-like cells(iCMs)by Sendai virus(SeV)vector overexpressing Gata4,Mef2c and Tbx5(SeVGMT).Methods CFs were divided into Control,NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh groups.NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh were transfected into CFs using Lipofectamine 3000 reagent for 48 hours.Lipofectamine 3000 reagent was then mixed with SeVGMT and incubated at room temperature for 48 hours,the culture medium was then replaced,and cells were cultured for 21 days.Cell morphology was observed under a microscope at 0,3,7,14,and 21 days.Expression levels of the myocardial-specific proteins α-myosin heavy chain(α-MHC),α-actin,cardiac troponin T(cTnT),connexin 43(Cx43),cardiac muscle α-actin(Actc1),and myosin heavy chain 6(Myh6)were detected at different time points by immunofluorescence,quantitative reverse transcription-polymerase chain reaction,and Western blot,and the proportions of beating cells at different time points were calculated.Results The relative fluorescence intensity and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs in each group increased with increasing culture time(P＜0.05),with significantly higher expression levels of myocardial-specific proteins at 14 days of culture than at 7 days(P＜0.05).The relative fluorescence intensities and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs at 3,7,14,and 21 days of culture were significantly increased in the CTRP3-Lv group compared with the NC-Lv group(P＜0.05),but were significantly decreased in CFs in the CTRP3-sh group compared with the NC-sh group(P＜0.05).Beating cells appeared in CFs in each group at 7 days of culture.The proportion of beating cells in each group increased with increasing culture time(P＜0.05),and the proportion was significantly higher at 14 days than at 7 days(P＜0.05).The proportion of beating cells among CFs was increased in the CTRP3-Lv group at 7,14,and 21 days of culture compared with the NC-Lv group(P＜0.05),while the proportion of beating cells in the CTRP3-sh group was decreased compared with the NC-sh group(P＜0.05).Conclusions CTRP3 can enhance SeVGMT reprogramming of CFs into iCMs.

Objective To establish the HPLC analysis method for ketoconazole related substances,and to provide technical support for the improvement of the quality standard and the purity of ketoconazole.Methods Gradient elution conditions were optimized based on the chromatographic parameters outlined in the 2024"British Pharmacopoeia"for ketoconazole cream.A C18 column(4.6 mm×150 mm,3 μm)was used to facilitate the elution process using a gradient of acetonitrile and acetate buffer,and detection performed at a wavelength of 230 nm.Results The known impurities and potentially genotoxic impurities in the material drugs of ketoconazole can be more effectively separated using the established methods,which are both robust and highly sensitive.The content of the two potentially geno toxic impurities,six known impurities and unknown impurities in the 21 batches of ketoconazole sourced from five raw material pharmaceutical companies is below the specified limit requirements.Conclusion This method contributes to improving the quality of ketoconazole ingredients,ensuring the safety of ketoconazole preparations,and better supporting the regulatory supervision.

Objective:To analyze the effects of Yiqi Tongluo Decoction combined with acupuncture on hemorheology and neurological function in patients with ischemic stroke due to qi deficiency and blood stasis syndrome. Methods:This study was designed as a prospective study. It included 120 patients with ischemic stroke due to qi deficiency and blood stasis syndrome, all of whom received treatment at Sun Simiao Hospital, Beijing University of Chinese Medicine between April 2021 and May 2023. The patients were randomly assigned to either an observation group ( n = 60) or a control group ( n = 60) using a random number table method. Both groups received standard conventional treatment. The control group received acupuncture, while the observation group was treated with both acupuncture and Yiqi Tongluo Decoction. Neurological function deficits were evaluated for both groups before and after treatment, and hemorheological indices, antioxidant enzyme levels, and markers of brain injury were measured to evaluate treatment efficacy. Results:After treatment, the scores on the China Stroke Scale and the National Institutes of Health Stroke Scale in the observation group were (11.37 ± 1.25) points and (6.18 ± 0.73) points, respectively, which were significantly lower than those in the control group [ (15.31 ± 1.59) points, (8.31 ± 0.92) points, t = 15.09,14.05, both P < 0.001]. After treatment, the plasma and whole blood viscosity levels in the observation group were (1.41 ± 0.15) mPa·s and (9.05 ± 1.01) mPa·s, respectively, which were significantly lower than those in the control group [(1.75 ± 0.18) mPa·s, (10.28 ± 1.14) mPa·s, t = 11.24,6.26, both P < 0.001]. The levels of serum catalase and glutathione peroxidase in the observation group were (160.85 ± 17.11) U/L and (40.31 ± 4.08) U/L, respectively, which were significantly higher than those in the control group [(139.64 ± 14.52) U/L, (34.85 ± 3.61) U/L, t = -7.32,-7.76, both P < 0.001]. The serum levels of high mobility group protein-1 and neuron-specific enolase in the observation group were (4.25 ± 0.43) μg/L and (18.57±1.92) μg/L, respectively, which were significantly lower than those in the control group [(5.84±0.61) μg/L, (22.63±2.31) μg/L, t = 10.47, 16.50, both P < 0.001]. The overall response rate in the observation group was significantly higher than that in the control group ( χ2 = 6.51, P < 0.05). Conclusions:Yiqi Tongluo Decoction combined with acupuncture has demonstrated significant benefits in treating patients with ischemic stroke due to qi deficiency and blood stasis syndrome. This approach can enhance hemorheology, increase antioxidant enzyme levels, minimize brain tissue damage, and improve neurological function in patients.

Objective To determine the diagnostic criteria of quantitative syndrome differentiation of toxin syndrome of diabetic kidney disease.Methods The questionnaire scale was developed through literature research and expert consultation.Points were assigned for the 5 major symptoms in 294 patients with DKD,and according to the TCM syndrome differentiation standard of toxic syndrome syndrome formulated by experts,it is divided into toxic syndrome group and non-toxic syndrome group.The symptom items were screened from the aspects of sensitivity,differentiation and representativeness by statistical method,and the weight value of the items was given by factor analysis.The threshold and the best diagnosis model were determined under the ROC curve.Finally,through the verification group data to verify the scale model,evaluate the diagnostic ability of the scale,and finally construct the diagnostic standard scale model of DKD toxin syndrome.Results 14 symptom items were selected as TCM related symptoms of DKD toxin syndrome,and the diagnostic threshold was determined to be 140.The diagnostic criteria of quantitative syndrome differentiation of DKD toxin syndrome were as follows:total score=fatigue * 10+edema * 10+turbid urine * 10+sore waist and knees * 10+dizziness * 10+tongue purple * 10+dark complexion * 9+limb numbness * 8+loose stools * 7+dry mouth * 4+dry eyes * 4+frequent urination at night * 3+abdominal distension * 3+greasy moss * 3.The degree of each item without this symptom should be multiplied by weight value by 0,mild by weight by 1,moderate by weight by 2,severe by weight by 3,and the total score≥140 could be diagnosed as toxin syndrome.The verification results showed that the sensitivity of the study group was 92.24%,the specificity was 96.19%,the Kappa value was 0.882,and the sensitivity,specificity and Kappa value of the verification group were 87.50%,96.97%and 0.836,respectively.Conclusion The standard scale of DKD toxin syndrome differentiation and diagnosis is constructed,and it has good diagnostic ability,which provides certain application value for clinical and scientific research.

Objective To determine the diagnostic criteria of quantitative syndrome differentiation of toxin syndrome of diabetic kidney disease.Methods The questionnaire scale was developed through literature research and expert consultation.Points were assigned for the 5 major symptoms in 294 patients with DKD,and according to the TCM syndrome differentiation standard of toxic syndrome syndrome formulated by experts,it is divided into toxic syndrome group and non-toxic syndrome group.The symptom items were screened from the aspects of sensitivity,differentiation and representativeness by statistical method,and the weight value of the items was given by factor analysis.The threshold and the best diagnosis model were determined under the ROC curve.Finally,through the verification group data to verify the scale model,evaluate the diagnostic ability of the scale,and finally construct the diagnostic standard scale model of DKD toxin syndrome.Results 14 symptom items were selected as TCM related symptoms of DKD toxin syndrome,and the diagnostic threshold was determined to be 140.The diagnostic criteria of quantitative syndrome differentiation of DKD toxin syndrome were as follows:total score=fatigue * 10+edema * 10+turbid urine * 10+sore waist and knees * 10+dizziness * 10+tongue purple * 10+dark complexion * 9+limb numbness * 8+loose stools * 7+dry mouth * 4+dry eyes * 4+frequent urination at night * 3+abdominal distension * 3+greasy moss * 3.The degree of each item without this symptom should be multiplied by weight value by 0,mild by weight by 1,moderate by weight by 2,severe by weight by 3,and the total score≥140 could be diagnosed as toxin syndrome.The verification results showed that the sensitivity of the study group was 92.24%,the specificity was 96.19%,the Kappa value was 0.882,and the sensitivity,specificity and Kappa value of the verification group were 87.50%,96.97%and 0.836,respectively.Conclusion The standard scale of DKD toxin syndrome differentiation and diagnosis is constructed,and it has good diagnostic ability,which provides certain application value for clinical and scientific research.

Objective:To investigate the clinical characteristics and influencing factors of secondary autoimmune encephalitis (AE) in adult patients with epidemic encephalitis B (EEB).Methods:78 adult EEB patients admitted to the Hanzhong Central Hospital from January 2006 to September 2017 were selected and divided into two groups based on the occurrence of secondary AE: group A (with secondary AE) and group B (without secondary AE). Factors influencing secondary AE in EEB patients were analyzed, and the receiver operating characteristic (ROC) curve was used to evaluate the predictive value of each factor for secondary AE.Results:After treatment, 28 out of 78 EEB patients (35.90%) developed secondary AE. The proportions of pulmonary infection, time from onset to admission, neutrophil-to-lymphocyte ratio (NLR), consciousness disorders, and respiratory disorders in the group A were significantly higher than those in the group B (all P<0.05). Binary logistic regression analysis showed that NLR, time from onset to admission, consciousness disorders, pulmonary infection, and respiratory disorders were influencing factors for secondary AE in EEB patients (all P<0.05). ROC curve analysis revealed that the combined detection of NLR, time from onset to admission, consciousness disorders, pulmonary infection, and respiratory disorders had higher sensitivity, specificity, and area under the curve (AUC) for predicting secondary AE than single detection (all P<0.05). Conclusions:The combined prediction of NLR, time from onset to admission, consciousness disorders, pulmonary infection, and respiratory disorders has high value for secondary AE in EEB patients. Clinically, these indicators can be used for early evaluation, and targeted interventions can be timely implemented to reduce the risk of secondary AE.

Objective The effect of intra-aortic balloon counterpulsation combined with recombinant human brain natriuretic peptide(rhBNP)treatment on ventricular remodeling after percutaneous coronary intervention(PCI)in elderly patients with acute anterior wall myocardial infarction.Methods A retrospective analysis was con-ducted on 66 elderly patients with acute anterior myocardial infarction who underwent emergency PCI in our hospital from January 2018 to June 2023.They were divided into a control group(n=32)who underwent PCI+rhBNP,and a study group(n=32)who underwent PCI+rhBNP+intra-aortic balloon counterpulsation.Blood pressure,lactic acid level,mechanical complications,ventricular remodeling,inflammatory factors,myocardial enzymology,ox-LDL level were detected and compared between the two groups.Results Before treatment,Lactic acid,diastolic blood pressure,systolic blood pressure,left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),left ventricular posterior wall thickness(LVPWT),left ventricular remodeling index(LVRI),left ventricular ejection fraction(LVEF),homocysteine(Hcy),interleukin-6(IL-6)and myocardial troponin were compared between the two groups.Myocardial troponin T(cTnT),lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),oxidized low density lipoprotein(ox-LDL)levels(P>0.05);At 3 months follow-up after discharge,the levels of lactic acid,LVESD,LVEDD,LVPWT,LVRI,Hcy,IL-6,cTnT,LDH,CK-MB and ox-LDL were decreased,and the levels were lower in the study group,and the levels of diastolic blood pressure,systolic blood pressure and LVEF were increased,and the levels were higher in the study group(P<0.05).The pro-portion of mechanical complications between the two groups before and after treatment were compared(P>0.05).Conclusion In elderly patients with acute anterior myocardial infarction after emergency PCI,intra-aortic balloon counterpulsation combined with rhBNP could alleviate ventricular remodeling,myocardial injury,reduce inflamma-tory injury,improve oxidative stress response,and regulate tissue perfusion.

Objective The effect of intra-aortic balloon counterpulsation combined with recombinant human brain natriuretic peptide(rhBNP)treatment on ventricular remodeling after percutaneous coronary intervention(PCI)in elderly patients with acute anterior wall myocardial infarction.Methods A retrospective analysis was con-ducted on 66 elderly patients with acute anterior myocardial infarction who underwent emergency PCI in our hospital from January 2018 to June 2023.They were divided into a control group(n=32)who underwent PCI+rhBNP,and a study group(n=32)who underwent PCI+rhBNP+intra-aortic balloon counterpulsation.Blood pressure,lactic acid level,mechanical complications,ventricular remodeling,inflammatory factors,myocardial enzymology,ox-LDL level were detected and compared between the two groups.Results Before treatment,Lactic acid,diastolic blood pressure,systolic blood pressure,left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),left ventricular posterior wall thickness(LVPWT),left ventricular remodeling index(LVRI),left ventricular ejection fraction(LVEF),homocysteine(Hcy),interleukin-6(IL-6)and myocardial troponin were compared between the two groups.Myocardial troponin T(cTnT),lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),oxidized low density lipoprotein(ox-LDL)levels(P>0.05);At 3 months follow-up after discharge,the levels of lactic acid,LVESD,LVEDD,LVPWT,LVRI,Hcy,IL-6,cTnT,LDH,CK-MB and ox-LDL were decreased,and the levels were lower in the study group,and the levels of diastolic blood pressure,systolic blood pressure and LVEF were increased,and the levels were higher in the study group(P<0.05).The pro-portion of mechanical complications between the two groups before and after treatment were compared(P>0.05).Conclusion In elderly patients with acute anterior myocardial infarction after emergency PCI,intra-aortic balloon counterpulsation combined with rhBNP could alleviate ventricular remodeling,myocardial injury,reduce inflamma-tory injury,improve oxidative stress response,and regulate tissue perfusion.

Objective:To analyze and summarize clinical phenotypic characteristics and genetic variations in patients with intellectual disability and pathogenic variations of the DYNC1H1 gene across 4 generations within a single family. Methods:Retrospective case analysis.Clinical data of a child with epilepsy and intellectual disability and her family members were collected from the Children′s Medical Center, Peking University First Hospital on December 2019.The child was followed up regularly.DNA was extracted from the peripheral blood of the child′s family members.Then whole-exome sequencing and Sanger sequencing were performed to identify the genetic variation type in the proband and her family members.The relationship between genotype and phenotype was further analyzed.Results:A total of 13 patients across 4 generations in the family had intellectual disability, and the proband also had drug-resistant epilepsy.The variation c. 13556C> A (p.A4519E) of the DYNC1H1 gene was confirmed by gene testing in 8 patients (no blood samples were obtained from the remaining patients). Conclusions:DYNC1H1 gene-related intellectual disability in most previously reported cases are caused by novel variations of this gene.In this study, a large family of 13 intellectual disability patients across 4 generations caused by a pathogenic mutation in the DYNC1H1 gene was summarized.The findings make precise genetic counseling possible for this family and provide a basis for further studies on the relationship between the genotype and phenotype of the DYNC1H1 gene.

BACKGROUND:Spinal canal decompression using uniportal endoscopic surgery is a new minimally invasive surgery in the treatment of lumbar spinal stenosis.However,this technique needs a steep learning curve and high requirements for surgical equipment and instruments,which limits its clinical application.We previously use the spinal endoscopy as a monitoring endoscopy and combined with unilateral biportal endoscopy to propose a hybrid technique of spinal endoscopy to achieve coaxial endoscopic operation and hands-separate operation. OBJECTIVE:To compare the clinical outcome of hybrid technique and uniportal endoscopic surgery in treatment of lumbar spinal stenosis with bilateral lower limb pain symptoms. METHODS:Ninety patients diagnosed of lumbar spinal stenosis with bilateral symptoms were included and retrospectively analyzed at First People's Hospital,Shanghai Jiao Tong University from August 2020 to August 2022.44 cases were included in group A(hybrid technique group),while 46 cases were included in group B(uniportal endoscopic surgery).The nerve decompression was observed during the surgery.Operation time,hospital stay time,and expenses were recorded in both groups.The visual analog scale scores of lower back pain and both lower extremities pain,Oswestry disability index scores of quality of life and excellent and good rate of modified Macnab criteria were recorded and compared at preoperative,postoperative 3 days,and postoperative 3 and 6 months. RESULTS AND CONCLUSION:(1)The operation time of group A was significantly shorter than that of group B(P<0.05).(2)The lower back pain and lower extremity pain of the severe side at postoperative 3 days,and 3 and 6 months were significantly relieved in both groups(P<0.05).The visual analog scale score of lower extremity pain on the mild side was significantly decreased at postoperative 3 days,3 and 6 months than preoperative score in the group A(P<0.05).The visual analog scale score of lower extremity pain on the mild side was significantly decreased at postoperative 3 days than preoperative score in the group B(P<0.05).The visual analog scale scores of lower extremity pain on the mild side at postoperative 3 and 6 months did not show significant difference than preoperative score in the group B.The comparison between the two groups showed that there was no significant difference in the visual analog scale scores of postoperative lower back pain and lower extremity pain of the severe side(P>0.05).The visual analog scale scores of lower extremity pain on the mild side in the group A were significantly lower than those of group B at postoperative 3 and 6 months(P<0.05).(3)The Oswestry disability index scores of both groups at postoperative 3 day were significantly lower than preoperative score(P<0.05),and there was no significant difference between the two groups 3 days after operation.Oswestry disability index scores of group A at postoperative 3 and 6 months were significantly decreased than preoperative score(P<0.05).The Oswestry disability index scores of group B at postoperative 3 and 6 months did not show significant differences than preoperative score(P>0.05).The comparison between the two groups showed the Oswestry disability index scores of group A were significantly lower than group B at postoperative 3 and 6 months(P<0.05).(4)The results of modified Macnab showed that the excellent and good rate of group A was significantly higher than that of group B(95%,78%,P<0.05).(5)It is indicated that the hybrid technique is a new spinal endoscopy technique,which has the advantages of less trauma and faster recovery as a minimally invasive surgery.The clinical outcome of hybrid technique is superior to that of uniportal endoscopic surgery in the treatment of lumbar spinal stenosis with bilateral symptoms.Additionally,it also has the advantages of good operational flexibility and high decompression efficiency as an open surgery.