In this study, we constructed a GLP-2R-HEK293 cell line and established a method for the determination of the in vitro biological activity of teduglutide based on HTRF, after optimizing experimental conditions and methodological verification. We also carried out relative potency detection of teduglutide pharmaceutical products using this method. The result showed that there was a quantitative-efficient relationship between the teduglutide activity and cAMP contents in GLP-2R-HEK293 cells, which conformed to four-parameter model. Method verification results of five concentrations of teduglutide (64%, 80%, 100%, 125% and 156%) met the requirements of the General Rules of Chinese Pharmacopoeia, 2020 edition, Volume IV (9401). We then analyzed the relative potency of three batches of teduglutide drug substances and three batches of drug products. The linearity, regression and parallelism of the obtained curves all fit the system suitability requirements. The relative potency of six batches of teduglutide was from 83% to 105%. In summary, the biological activity detection method established in this study was accurate, precise, simple and time-saving, which can be used for quality control of teduglutide pharmaceutical products.

Objective To analyze the structural and phylogenetic characteristics of the mitochondrial genome from Bulinus globosus, so as to provide a theoretical basis for classification and identification of species within the Bulinus genus, and to provide insights into understanding of Bulinus-schistosomes interactions and the mechanisms of parasite transmission. Methods B. globosus samples were collected from the Ruya River basin in Zimbabwe. Mitochondrial DNA was extracted from B. globosus samples and the corresponding libraries were constructed for high-throughput sequencing on the Illumina NovaSeq 6000 platform. After raw sequencing data were subjected to quality control using the fastp software, genome assembly was performed using the A5-miseq and SPAdes tools, and genome annotation was conducted using the MITOS online server. Circular maps and sequence plots of the mitochondrial genome were generated using the CGView and OGDRAW software, and the protein conservation motifs and structures were analyzed using the TBtools software. Base composition and codon usage bias were analyzed and visualized using the software MEGA X and the ggplot2 package in the R software. In addition, a phylogenetic tree was created in the software MEGA X after sequence alignment with the software MAFFT 7, and visualized using the software iTOL. Results The mitochondrial genome of B. globosus was a 13 730 bp double-stranded circular molecule, containing 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and 13 protein-coding genes, with a marked AT preference. The mitochondrial genome composition of B. globosus was similar to that of other species within the Bulinus genus. Phylogenetic analysis revealed that the complete mitochondrial genome sequence of B. globosus was clustered with B. truncatus, B. nasutus, and B. ugandae into the same evolutionary clade, and gene superfamily analysis showed that the metabolism-related proteins of B. globosus were highly conserved, notably the cytochrome c oxidase family, which showed a significant consistency. Conclusions This is the first whole mitochondrial genome sequencing to decode the compositional features of the mitochondrial genome of B. globosus from Zimbabwe and its evolutionary relationship within the Bulinus genus, which provides important insights for further understanding of the phylogeny and mitochondrial genome characteristics of the Bulinus genus.

ObjectiveTo analyze the risk factors of dementia among healthy elderly individuals in the middle of their lives. MethodsA total of 175 participants aged 55 to 75 from two communities in Beijing were included from July, 2021 to April, 2023. Cardiovascular Risk Factors, Aging, and Incidence of Dementia (CAIDE) related risk factors and other demographic data were collected. According to the CAIDE assessment, participants with scores ≥ 9 were as high-risk group, and those with scores < 9 were as low-risk group. They were evaluated with Stroop Color Word Test (SCWT), two elements 1-back task paradigm and the revised Trail Making Test (TMT); measured the grip strength, 30 s forearm flexion tests and five sit-to-stand tests; the average step speed and step length of a 10-meter walk were recorded. ResultsThe average total score of CAIDE was 9.86 in the high-risk group, and was 4.95 in the low-risk group. There was no difference in age between two groups (P = 0.188). There were differences in the proportion of participants of male, less than seven years' education, systolic blood pressure > 140 mmHg, cholesterol > 6.5 mmol/L, body mass index > 30 kg/m², and lack of physical activity between two groups (χ² > 3.116, P < 0.05). The grip strength (t = -4.174), walking speed (t = -2.414), SCWT accuracy (Z = -2.684) were all worse in the high-risk group than in the low-risk group (P < 0.05). Logistic regression analysis showed that walking speed (OR = 25.483), grip strength (OR = 1.133) and SCWT accuracy (OR = 37.430) were independent influencing factors of dementia (P < 0.05). ConclusionWeaker grip strength, slower gait speed and worse inhibitory control might be independent influencing factors of dementia.

This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL^-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.

Objective:There are a variety of minimally invasive interventional treatments for trigeminal neuralgia,and the efficacy evaluation is different.The preferred treatment scheme is still controversial.This study aims to investigate the differences in treatment effects between patients with primary trigeminal neuralgia(PTN)treated with percutaneous balloon compression(PBC)for the first intervention and patients with pain recurrence after radiofrequency thermocoagulation(RT)who then received PBC for PTN,and to offer clinicians and patients more scientifically grounded and precise treatment alternatives. Methods:We retrospectively analyzed 103 patients with PTN admitted to the Department of Pain Management of the Second Affiliated Hospital of Guangxi Medical University from January 2020 to December 2021,including 49 patients who received PBC for the first time(PBC group)and 54 patients who received PBC for pain recurrence after RT(RT+PBC group).General information,preoperative pain score,intraoperative oval foramen morphology,oval foramen area,balloon volume,duration of compression,and postoperative pain scores and pain recurrence at each time point on day 1(T1),day 7(T2),day 14(T3),1 month(T4),3 months(T5),and 1 year(T6)were collected and recorded for both groups.The differences in treatment effect,complications and recurrence between the 2 groups were compared,and the related influencing factors were analyzed. Results:The differences of general information,preoperative pain scores,foramen ovale morphology,foramen ovale area,T1 to T3 pain scores between the 2 groups were not statistically different(all P>0.05).The balloon filling volume in the PBC group was smaller than that in the RT+PBC group,the pain scores at T4 to T6 and pain recurrence were better than those in the RT+PBC group(all P<0.05).Pain recurrence was positively correlated with pain scores of T2 to T6(r=0.306,0.482,0.831,0.876,0.887,respectively;all P<0.01). Conclusion:The choice of PBC for the first intervention in PTN patients is superior to the choice of PBC after pain recurrence after RT treatment in terms of treatment outcome and pain recurrence.

Objective To analyze the correlation between sympathetic nerve excitability and the occurrence and development of wake-up stroke.Methods A total of 100 patients with acute stroke admitted to our hospital from July 2020 to January 2022 were selected and divided into wake-up stroke group and non-wake-up stroke group according to whether wake-up stroke occurred.Logistic regression model was established to analyze the relationship between sympathetic excitation and the occurrence and development of wake-up stroke.Results Multivariate analysis showed that blood pressure variability,heart rate variability,blood glucose variability,electrolytes disturbance,25-hydroxyvitamin D3,urinary vanilla bitter almond acid,and catecholamine were risk factors for the occurrence and development of stroke after waking up(all P＜0.05).Conclusions Sympathetic nerve excitability is related to the occurrence and development of stroke after waking up.Blood pressure variability,heart rate variability,blood glucose variability,electrolyte disturbance,25-hydroxyvitamin D3,urinary vanilla bitter almond acid and catecholamine are risk factors for the occurrence and development of stroke after waking up.

Objective To explore clinical value of the expression levels of cell surface transmembrane glycoprotein molecule 44(CD44)mRNA,cell surface transmembrane glycoprotein molecule 24(CD24)mRNA,and protein in the placenta of severe preeclampsia(SPE)patients.Methods The SPE patients who were delivered by cesarean section in the Second People's Hospital of Liaocheng from June 2019 to June 2022 were further divided into 45 patients in early onset SPE group(gestational age≤34 weeks)and 55 patients in late onset SPE group(gestational age＞34 weeks)according to the different gestational age.The control group consisted of 100 normal cases in the same period.The expression of CD44 and CD24 in placenta of SPE patients was detected by fluorescent quantitative PCR and immunohistochemistry,Pearson method was used to analyze the difference of their expression levels and their correlation with the clinical characteristics of SPE disease,and multivariate logistic regression was used to analyze the influencing factors of SPE.Results Compared with the control group,the expression levels of CD44 mRNA(0.55±0.12 vs 1.02±0.33)and CD24 mRNA(0.68±0.19 vs 1.05±0.11)in SPE placental tissues decreased significantly,the differences were statistically significant(t=13.385,16.853,P＜0.05).The immunohistochemical staining results showed that CD44 and CD24 were mostly negative or weakly positive in the SPE group placental tissue,while they were mostly positive in the control group,the positive rates of CD44 and CD24 in the SPE placental tissue were lower than those in the control group,and the differences were statistically significant(χ2=9.696,14.346,P＜0.05).Compared to the early onset SPE group,the expression levels of CD44(0.65±0.17 vs 0.42±0.11)and CD24(0.77±0.23 vs 0.58±0.13)mRNA in placental tissue of late onset SPE were higher,and the differences were statistically significant(t=7.830,4.932,P＜0.05).Compared with the control group,the BMI,systolic blood pressure,diastolic blood pressure,urinary protein,Cr,LDH and BUN were significantly increased in SPE group(t=5.360～30.241,all P＜0.05).In SPE group,the gestational age was earlier,the MPV and ALB were lower,the newborn's birth length was shorter,and the body weight than control group,the differences were statistically great(t=3.232～11.109,all P＜0.05).The expression of CD44 and CD24 in SPE placenta was positively correlated(r=0.698,P＜0.05),the expression of CD44 in SPE placenta was positively correlated with CD24,gestational week of delivery,MPV and neonatal birth length(r=0.611,0.639,0.612,0.465,all P＜0.05),and was negatively correlated with systolic blood pressure,urinary protein and LDH(r=-0.604,-0.569,-0.593,all P＜0.05).The expression of CD24 was positively correlated with gestational age,MPV and newborn birth length(r=0.605,0.584,0.640,all P＜0.05),and was negatively correlated with systolic blood pressure,urinary protein and LDH(r=-0.637,-0.593,-0.561,all P＜0.05).The results of logistic regression analysis showed that MPV(95％CI:1.429～4.350),urinary protein(95％CI:1.529～2.709),and LDH(95％CI:1.425～3.932)were all independent risk factors for SPE(all P＜0.05).High levels of CD44(95％CI:0.561～0.940)and CD24(95％CI:0.495～0.814)were independent protective factors for SPE(P＜0.05).Conclusion The low expression levels of CD44 and CD24 in placenta of SPE patients are independent protective factors of SPE,which can provide direction for the follow-up treatment of SPE.

Objective:To explore the application of a role transition shock model based on the teach-back technique in knee arthroplasty nursing teaching.Methods:We assigned 50 nursing student interns practicing in the knee arthroplasty team of Orthopedics Department of Nanjing First Hospital between August 2020 and August 2022 into control group ( n=25, traditional teaching) and observation group ( n=25, teach-back-based role transition shock model teaching) according to the order of admission. At the end of internship, the examination scores, the impact of transition shock on comprehensive abilities, and teaching satisfaction of the students were assessed and analyzed using the t test and Fisher's exact test with the use of SPSS 22.0. Results:Compared with the control group, the observation group scored significantly lower in the physical, psychological, knowledge and skills, and sociocultural and developmental dimensions of the transition shock assessment scale ( P<0.05). The observation group showed significantly higher scores of nurse-patient communication, nursing practice, disease observation, health education, humanistic care, team cooperation, clinical thinking, and emergency response than the control group ( P<0.05). The examination results of the observation group were significantly better than those of the control group ( t=12.31, 11.52, P<0.001). The teaching satisfaction rate of the observation group was significantly higher than that of the control group [100.00% (25/25) vs. 68.00% (17/25), χ2=9.52, P=0.002]. Conclusions:The teach-back-based role transition shock model can help alleviate the transition impact faced by nursing student interns when entering clinical practice, and also improve their comprehensive abilities as well as satisfaction with teaching.

AIM:To study whether glycyrrhizic acid(GL)can resist the ototoxicity of cisplatin(CDDP)in mice and its molecular mechanism.METHODS:Male C57BL/6J mice were divided into 5 groups:control group,DMSO(5%)group,CDDP(4 mg/kg)group,CDDP+low-dose(50 mg/kg)GL group,and CDDP+high-dose(100 mg/kg)GL group(n=14).Auditory brainstem response(ABR)was used to detect hearing changes of mice.HE staining was used to observe the morphological change of cochlear stria vascular in mice.Evans blue(EB)staining was used to observe the per-meability change of the blood-labyrinth barrier(BLB).Immunohistochemical technique was used to detect the expression and distribution of adhesion protein VE-cadherin and tight junction protein ZO-1 on the cochlear stria.ELISA assay and immunofluorescence technology were employed to detect the expression of tumor necrosis factor-α(TNF-α)and interleu-kin-1β(1L-1β).RESULTS:In CDDP group,ABR waveforms of all frequencies were disturbed,the hearing threshold was significantly increased,and I wave latency was prolonged(P＜0.05).In CDDP+GL group,ABR waveforms of various frequencies were well differentiated,the hearing threshold was significantly decreased,and the latency of I-wave was shortened(P＜0.01).The disordered morphology and more vacuoles in the stria vascularis were observed by HE staining in CDDP group.The GL alleviated CDDP-induced damage in the stria vascularis.In EB staining,CDDP caused an increase in per-meability of BLB(P＜0.01),which was improved by GL treatment(P＜0.01).Immunohistochemical results showed that the expression of VE-cadherin and ZO-1 in CDDP group were decreased(P＜0.01),which was restored in CDDP+GL group(P＜0.01).The ELISA and immunofluorescence results showed that the expression of IL-1β and TNF-α was in-creased after CDDP treatment(P＜0.01),which was restored in CDDP+GL group(P＜0.01).CONCLUSION:The GL alleviates CDDP-induced hearing loss in mice by inhibiting CDDP-induced inflammation and reducing the permeability of BLB.

Objective To compare the therapeutic efficacy of whey protein powder in peritoneal dialysis hypoproteinemia patients.Methods From August 2019 to August 2020,100 patients with peritoneal dialysis hypoproteinemia from the Peritoneal Dialysis Center of the First Affiliated Hospital of Xi'an Jiaotong University were investigated.The patients were separated into 2 groups randomly;the study group were administered whey protein powder and the control group with high protein food.After 32 weeks of treatment,biochemical and biometric indices including hemoglobin(Hb),albumin(ALB),prealbumin(PA),triglyceride(TG),total cholesterol(TC),phosphorus(P),low density lipoprotein(LDL),high density lipoprotein(HDL),serum creatinine(Scr),blood urea nitrogen(BUN),estimated glo-merular filtration rate(eGFR),total spKt/Vurea(TKt/V),total creatinine clearance rate(TCcr),hand grip strength(HG),triceps skinfold(TSF),arm circumference(AMC),mid-arm muscle circumference(MAMC)were compared between groups.Results Compared with 0 week,at 16 and 32 weeks,ALB,PA,and HG were significantly increased in the study group(P＜0.05).Compared with the control group,ALB,PA,and HG increased significantly at 16 and 32 weeks in the study group(P＜0.05).There were no significant differences in TG,TC,HDL,LDL,eGFR,TKt/V,and TCcr at 0,8,16,and 32 weeks between the control and study groups(P＞0.05).Conclusion For patients with peritoneal dialysis hypoproteinemia caused by insufficient protein intake or excessive protein loss,the addition of whey protein during peritoneal dialysis can significantly improve the nutritional status of patients,with greater efficacy than a high protein diet alone.