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.

Aim To observe the interventional effects of Polygala fallax Hemsl.(PFH)on diabetic kidney disease(DKD)rats and the regulatory mechanism of ferroptosis.Methods Thirty-six SD rats were ran-domly divided into the control group,DKD group,fer-rostatin-1(Fer-1)group,PFH-L group,PFH-M group,and PFH-H group,with six rats in each group.Model-ing was induced by a one-time intraperitoneal injection of 35 mg·kg-1 streptozocin(STZ)in combination with high-sugar and high-fat feed.Ferrostatin-1(25μmol·kg-1)was injected intraperitoneally in Fer-1 group.The PFH-L,PFH-M,and PFH-H groups were gavaged with 50,100 and 200 mg·kg-1 of alcoholic extracts of PFH respectively,and the control and DKD groups were gavaged with an equal volume of distilled water once a day for eight weeks.At the end of drug administration,blood glucose,24h-UP,BUN and Scr levels were measured in each group of rats.HE stai-ning and Masson staining were used to observe renal histopathological changes.ELISA was employed to de-termine the levels of total iron,MDA and GSH activity.IHC was used to observe the expression of Nrf2,SLC7A11,and GPX4 in renal tissues.Western blot was used to detect the protein expression of COL1A1,α-SMA,TGF-,FTH-1,TFR-1,Nrf2,SLC7A1 1,GPX4,in renal tissues.RT-qPCR was used to detect the ex-pression levels of Nrf2,SLC7A11,GPX4 mRNA in re-nal tissues.Results Compared with the control group,blood glucose,24h-UP,BUN,Scr increased(P＜0.05);glomerular volume increased,tubular dilata-tion and collagen fiber deposition were obvious;total i-ron,MDA content increased and GSH activity de-creased(P＜0.05);there was increased protein ex-pression of COL1A1,α-SMA,TGF-β,TFR-1 and de-creased protein expression of FTH-1(P＜0.01);there was decreased mRNA and protein expression of Nrf2,SLC7A11,GPX4 in DKD group rats(P＜0.01).Compared with the DKD group,blood glucose,24h-UP,BUN and Scr decreased(P＜0.05);renal tissue le-sions were significantly reduced;total iron and MDA content decreased,and GSH activity increased(P＜0.05);COL1A1,α-SMA,TGF-β,and TFR-1 protein expression decreased and FTH-1 protein expression in-creased(P＜0.05,P＜0.01);Nrf2,SLC7A11,GPX4 mRNA and protein expression increased in Fer-1 and PFH dose groups(P＜0.05,P＜0.01).Conclusions PFH attenuates renal histopathological injury in DKD rats,and the mechanism may be related to the regula-tion of the Nrf2/SLC7A11/GPX4 signaling pathway.

Purpose To explore the clinicopathological features,diagnosis and differential diagnosis of mantle cell lymphoma(MCL)with aberrant expression of CD10.Methods 14 cases of MCL with aberrant expression of CD10 were analyzed using hematoxylin-eosin,immunohistochemical stains,in situ hybridization,and fluorescence in situ hy-bridization(FISH)techniques to observe the histological morphology,immunophenotype,and molecular genetic char-acteristics.The relevant literatures were reviewed.Results There were 11 males and 3 females,with a male-to-fe-male ratio of 11∶3.The age ranged from 49 to 80 years,with an average age of 64.4 years and a median age of 64 years.10 cases occurred in lymph nodes,1 case in the nasopharynx,1 case in the right colon,1 case in the right eye-lid,and 1 case in the right testis.According to the Ann Arbor staging system,8 cases were classified as stage Ⅳ and 5 cases as stage Ⅲ,and 1 case with undetermined staging.Histologically,there was diffuse effacement of the normal architecture by tumor cells infiltration.Transparent degenerate small blood vessels and scattered individual epithelial-like tissue cells could be observed in the background.Among them,8 cases(8/14,57.14％)were composed of uni-form small to medium-sized lymphocytes with slightly irregular nuclei,unevenly dispersed chromatin,inconspicuous nucleoli,and scant cytoplasm,along with observable mitotic figures.In 3 cases(3/14,21.43％),the tumor cells were large and markedly pleomorphic,with round or irregular nuclei,prominent nucleoli,frequent mitotic figures,and abundant pale cytoplasm.Tumor cells in 3 cases(3/14,21.43％)were resembling lymphoblasts,characterized by round nuclei,fine chromatin,inconspicuous nucleoli,frequent mitotic figures,and scant cytoplasm.Immunophenotyp-ically,CD21 staining showed residual follicular dendritic meshworks.The tumor cells were diffusely and strongly posi-tive for CD20(14/14),PAX5(7/7),CD5(14/14),Cyclin D1(14/14),SOX11(11/11),and BCL2(13/13),partially positive for BCL6(8/14,57.14％)and MUM1(6/9,66.67％),but negative for CD3(14/14)and CD23(14/14).Among 14 cases,10 cases were diffusely and strongly positive for CD10(10/14,71.43％),and 4 cases were partially positive for CD10(4/14,28.57％).The percentage of Ki67 index ranged from 10％to 90％.All cases were negative for EBER(8/8).FISH analysis was performed in 9 cases,among which 7 cases showed CCND1 gene rearrangement,another 2 cases failed to detect due to insufficient tissue samples.Bone marrow biopsy was performed in 13 cases,revealing involvement in 8 cases(8/13,61.54％)and no involvement in 5 cases(5/13,38.46％).Con-clusion MCL with aberrant expression of CD10 is very rare,which commonly exhibits a diffuse growth pattern and blastoid and pleomorphic variants.It often has a high Ki67 proliferation index and poorer prognosis,and should be dis-tinguished from other subtypes of CD1O-positive B-cell lymphomas.

Objective:To investigate the clinical efficacy of cervical cerclage in singleton pregnancy with different degrees of short cervix.Methods:The clinical data of singleton pregnant women who underwent transvaginal ultrasound examination at 18-24 +6 weeks of gestation and found cervical dilation with cervical length (CL) ≤20 mm, and without history of spontaneous preterm delivery or late abortion in Women′s Hospital, Zhejiang University School of Medicine from January 2021 to September 2023 were collected and retrospectively analyzed. According to the case control matching, 78 pregnant women in the cerclage group and 78 women in the conservative treatment group were finally included. The pregnancy outcomes and neonatal prognosis of the two groups were compared. Meanwhile, the two groups of pregnant women were divided into three subgroups for stratification (CL≤10, 11-15, 16-20 mm). Multivariate logistic regression analysis and Kaplan-Meier curve were used to evaluate the effect of cervical cerclage on pregnancy outcomes in different subgroups. Results:(1) Compared with the conservative treatment group, the gestational age at delivery (median: 36 vs 37 weeks) and the duration of pregnancy extension (median: 90 vs 97 days) in the cerclage group were not significantly prolonged (all P>0.05). There was no significant difference in the cumulative non-delivery rate between the cerclage group and the conservative treatment group ( P=0.143). The rate of neonatal intensive care unit (NICU) admission in the cerclage group was significantly higher than that in the conservative treatment group (46.1% vs 31.2%, P<0.001), but there were no significant differences in other neonatal outcomes between the two groups (all P>0.05). (2) CL 16-20 mm subgroup: compared with the conservative treatment group (28 cases), the cerclage group (18 cases) had a significantly higher rate of preterm birth before 37 weeks of gestation (3.6% vs 6/18, P<0.001) and a significantly lower neonatal birth weight (median: 3 370 vs 2 925 g, P<0.001). There was no significant difference in the cumulative non-delivery rate between the cerclage group and the conservative treatment group ( P=0.168). (3) CL 11-15 mm subgroup: compared with the conservative treatment group (26 cases), the gestational age of delivery in the cerclage group (32 cases) was later (median: 36 and 37 weeks, respectively), and the difference was statistically significant ( P<0.05). The cumulative non-delivery rate in the cerclage group was significantly higher than that in the conservative treatment group ( P=0.001). (4) CL≤10 mm subgroup: compared with the conservative treatment group (24 cases), the pregnant women in the cerclage group (28 cases) had a later gestational age at delivery (median: 34 vs 37 weeks), a longer duration of pregnancy (median: 74 vs 97 days), and a larger newborn birth weight (median: 2 300 vs 3 165 g). The rates of preterm birth before 34 weeks of gestation (45.8% vs 14.3%) and before 37 weeks of gestation (83.3% vs 39.3%) were lower, and the differences were all statistically significant (all P<0.05). The cumulative non-delivery rate of pregnant women in the cerclage group was significantly higher than that in the conservative treatment group ( P<0.001). Conclusion:Cervical cerclage could significantly prolong the gestational weeks and improve the perinatal outcomes for singleton pregnant women with cervical internal orifice dilation and CL≤15 mm without a history of spontaneous preterm delivery or late abortion.

Objective To investigate the effect of Modified Renshen Wumei Decoction on the TLR4/MyD88/pNF-κBp65 signaling pathway and elucidate the potential mechanism by which this formula repairs the intestinal mucosal barrier in diarrheal rats.Methods Twelve rats were randomly selected from a total of 48 rats to serve as the blank control group(CK),while the remaining 36 rats were used to establish a disease model via a compound method.After 14 days of model preparation,the rats were randomly divided into three groups:the model group(MC),the western medicine group(MV),and the traditional Chinese medicine group(MRWD).Each of the four groups(including CK)received corresponding interventions for 7 days.The concentrations of serum diamine oxidase(DAO),D-lactic acid(D-Lac),interleukin-1β(IL-1β),IL-6,IL-10,tumor necrosis factor-α(TNF-α),mucin 2(MUC2),MUC4,MUC6,and colonic homogenate secretory immunoglobulin A(SIgA)were measured using ELISA.Additionally,the protein and gene expressions of colonic toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),phosphorylated nuclear factor kappa-light-chain-enhancer of activated B cells p65(pNF-κBp65),occludin,claudin-1,and zonula occludens-1(ZO-1)were analyzed by Western blot and RT-PCR.Results(1)Intestinal mucosal injury markers:Compared with the blank group,the serum levels of DAO and D-Lac in the model group were significantly increased(P<0.05).Compared with the model group,both the Chinese medicine group and Western medicine group significantly decreased the serum levels of DAO(P<0.001),while the traditional Chinese medicine group also significantly reduced the serum levels of D-Lac(P<0.05).There was no significant difference in the changes of DAO and D-Lac serum levels between the Chinese medicine group and Western medicine group(P>0.05).(2)Inflammatory indicators:Compared with the blank group,the model group exhibited significant upregulation of TLR4,MyD88,pNF-κ Bp65 protein and gene expression,as well as serum levels of IL-1β,IL-6,and TNF-α(P<0.05),along with a significant decrease in IL-10 serum levels(P<0.05).Compared with the model group,both the Chinese medicine group and Western medicine group significantly downregulated TLR4,MyD88,pNF-κBp65 protein and gene expression,as well as serum levels of IL-1β,IL-6,and TNF-α(P<0.05),and significantly upregulated IL-10 serum levels(P<0.05).There was no significant difference in serum levels of TLR4,MyD88,pNF-κBp65 protein,gene expression,and IL-1β,IL-6,IL-10,and TNF-α between the Chinese medicine group and Western medicine group(P>0.05).(3)Intestinal mucosal barrier factors:Compared with the blank group,the model group exhibited significant downregulation in MUC2,MUC6,SIgA content,as well as Claudin-1,ZO-1 protein and gene expression,and Occludin protein expression(P<0.05).Compared with the model group,both Chinese and Western medicine groups significantly upregulated the content of MUC2 and SIgA,as well as the protein and gene expression of Claudin-1 and ZO-1(P<0.05).The traditional Chinese medicine group also significantly increased the content of MUC6 and Occludin protein expression(P<0.05).No significant differ-ences were observed between the Chinese and Western medicine groups in terms of MUC2,MUC6,SIgA serum content,and Claudin-1 and ZO-1 gene expression(P>0.05).However,the Western medicine group showed better Claudin-1 protein expression than the Chinese medicine group(P<0.05),while the ZO-1 protein expression was higher in the traditional Chinese medicine group compared to the Western medicine group(P<0.05).Conclusion Modified Renshen Wumei Decoction exerts an intestinal mucosal barrier repair effect in diarrhea rats by modulating the TLR4/MyD88/pNF-κBp65 signaling pathway.

Objective To explore the impact and prevention strategies of medical device adverse events in a children's hospital,and to improve the monitoring and management capabilities of medical device adverse events.Methods We retrieved 367 medical device adverse events that occurred in a children's hospital from 2017 to 2022.Using Excel software,we analyzed,organized,and statistically summarized the annual reports of adverse events,the distribution of event grades,the age distribution of involved patients,and the main medical devices involved.Results The annual reports of medical device adverse events showed an overall upward trend.Grade Ⅲ and Ⅳ adverse events accounted for the majority,representing 97.28％of the total re-ports.The main age group affected was 0-3 years old,accounting for 43.60％of the total adverse event reports.Disposable ster-ile syringes,disposable vacuum blood collection tubes,disposable gastric tubes,disposable infusion sets(with needles),dispos-able intravenous catheters,and other polymer products accounted for a high proportion of adverse events.Adverse events related to infusion,nursing,and protective equipment also accounted for a high proportion.Conclusion Adopting measures such as continuous monitoring and analysis of medical device adverse event reports,improving the construction of adverse event systems,strengthening professional skill training for medical staff,and enhancing quality management are conducive to improving the man-agement level of medical device adverse events in hospitals and ensuring patient safety.

Objective:To learn about the clinical manifestations, laboratory test results, and prognosis of patients with acute and chronic brucellosis, and to provide a scientific basis for clinical diagnosis and treatment.Methods:A retrospective study was conducted to collect the clinical data of 776 patients with brucellosis admitted to the First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2023. The patients were divided into acute phase group and chronic phase group according to the course of disease. Their clinical manifestations, laboratory test results, and prognosis were compared and analyzed between the two groups.Results:Out of 776 patients with brucellosis, 649 were in the acute phase group and 127 were in the chronic phase group. Male accounted for 71.6% (556/776). The age was (44.91 ± 0.60) years old, mainly concentrated in the age range of 46 - 60 years old (38.0%, 295/776). Farmers were the main occupation, accounting for 58.4% (453/776). The disease occurred in all months of the year, with autumn being the peak period (33.9%, 263/776). The primary clinical manifestations were fever (64.7%, 502/776) and fatigue (40.5%, 314/776). The incidence rates of fever and fatigue in the acute phase group were higher than those in the chronic phase group [70.4% (457/649) vs. 35.4% (45/127), 42.5% (276/649) vs. 29.9% (38/127)], with statistically significant differences ( χ2 = 56.91, 7.01, P < 0.05). Complications of the osteoarticular system were the most common, accounting for 47.8% (371/776). Abnormal results of blood routine examinations were mainly characterized by decreased hemoglobin, decreased lymphocytes, and decreased white blood cells, accounting for 53.7% (417/776), 32.6% (253/776), and 22.6% (175/776), respectively. The incidence rate of decreased hemoglobin in the acute phase group was higher than that in the chronic phase group [63.5% (412/649) vs. 3.9% (5/127)], with a statistically significant difference ( χ2 = 151.49, P < 0.001). The remaining abnormal laboratory test results were mainly characterized by elevated erythrocyte sedimentation rate, elevated C-reactive protein, and elevated aspartate aminotransferase, accounting for 75.1% (583/776), 50.6% (393/776), and 39.8% (309/776), respectively. The incidence rates of elevated erythrocyte sedimentation rate and elevated aspartate aminotransferase in the acute phase group were higher than those in the chronic phase group [77.8% (505/649) vs. 61.4% (78/127), 43.3% (281/649) vs. 22.0% (28/127)], with statistically significant differences ( χ2 = 15.28, 20.02, P < 0.001). The overall positive rate of Brucella blood culture was 57.5% (446/776), and there was a statistically significant difference in blood culture between the two groups ( χ2 = 17.08, P = 0.002). The positive rate of blood culture in the acute phase group was higher than that in the chronic phase group [60.1% (390/649) vs. 44.1% (56/127)]. Ninety-four point three percent (732/776) of patients were treated with antibiotics, with rifampicin + doxycycline as the main treatment regimen (45.2%, 331/732). The median of antibiotic types used in the acute and chronic phase groups were 3 and 4, respectively. The overall incidence rate of adverse drug reactions was 3.8% (28/732). Eighty-seven point five percent (625/714) of patients improved or recovered after treatment, while 12.5% (89/714) did not recover or experienced relapse. Conclusions:The main clinical manifestations of brucellosis patients are fever and fatigue, with a higher incidence of complications in the osteoarticular system, and a better prognosis. The incidence of fever, fatigue, decreased hemoglobin, elevated erythrocyte sedimentation rate, elevated aspartate aminotransferase, and positive Brucella blood culture in patients in the acute phase are higher, and the types of antibiotics used are fewer than those in patients in the chronic phase.

Objective To explore the clinical effects and influencing factors of recombinant human growth hormone(rhGH)treatment in pediatric patients with growth hormone deficiency.Methods The study subjects were pediatric patients with growth hormone deficiency,all of whom received a combination therapy of stanozolol tablets 2 mg(qd)and 0.1 U·kg-1·d-1 of recombinant human growth hormone injection for a continuous period of one year.After one year,the patients were divided into active group and invalid group based on clinical outcomes.Comparisons were made between the two groups in terms of height,annual growth velocity(GV)and height standard deviation score(HtSDS)before and after treatment,and safety evaluations were conducted.Multivariate Logistic regression analysis was used to identify factors that influenced the treatment outcomes in pediatric patients with growth hormone deficiency.Results After one year of treatment,a total of 93 cases pediatric patients were included in the analysis,with 62 cases in the active group and 31 cases in the invalid group.The heights of the patients before and after treatment were(119.40±2.48)and(129.08±2.37)cm,respectively;the GV were(3.08±0.39)and(7.34±1.02)cm·year-1,respectively;the HtSDS were-2.45±0.49 and-1.68±0.35,respectively,with statistically significant differences observed for all comparisons(all P＜0.05).In the active and invalid groups,the proportions of patients with GV＜3.0 cm·year-1 before treatment were 32.26％and 58.06％,respectively;the proportions of patients with peak growth hormone(GH)levels＜5.0 ng·mL-1 before treatment were 30.65％and 54.84％,respectively;the average maternal heights were(158.83±5.76)and(155.48±6.58)cm,respectively,with statistically significant differences observed for all comparisons(all P＜0.05).The results of the Logistic regression model showed that GV＜3.0 cm·year-1 before treatment,peak GH levels＜5.0 ng·mL-1 before treatment,and a shorter maternal height were independent risk factors for poor treatment outcomes with rhGH in pediatric patients with growth hormone deficiency(all P＜0.05).No severe adverse reactions occurred during the treatment of any patient;seven patients experienced pain at the injection site,one patient had a slight increase in blood glucose,and five patients had mild decreased appetite.The total incidence of adverse drug reactions were 13.98％(13 cases/93 cases).Conclusion Treatment with rhGH for pediatric patients with growth hormone deficiency can significantly improve height development and has good safety,but it is influenced by factors such as growth velocity,peak GH levels,and maternal height.

Objective To identify the pathogenic gene and variant for a family with branchio-otic syndrome.Methods The clinical data of this family were collected,and the peripheral blood was extracted for deafness gene NGS panel analysis.Pathogenic variation detected was verified by Sanger sequencing.Results The family contained 17 members in three-generations,3 of whom exhibited autosomal dominant,hearing loss,preauricular fistula and branchial cleft fistula,which were in accordance with the clinical diagnosis criteria of branchio-otic syndrome.A no-vel heterozygous variant c.963dupT(p.E322X)in EYA1 gene was identified,which co-segregated with the branchi-o-otic syndrome phenotype in the family.The variant was a nonsense variant resulting in the premature appearance of the stop codon.According to the American College of Medical Genetics and Genomics(ACMG)guidelines and the criteria,the variant was classified as pathogenic.Conclusion We identified a novel pathogenic variant EYA1:c.963dupT(p.E322X)in a family with branchio-otic syndrome.

Objective To identify the pathogenic gene and variant for a family with branchio-otic syndrome.Methods The clinical data of this family were collected,and the peripheral blood was extracted for deafness gene NGS panel analysis.Pathogenic variation detected was verified by Sanger sequencing.Results The family contained 17 members in three-generations,3 of whom exhibited autosomal dominant,hearing loss,preauricular fistula and branchial cleft fistula,which were in accordance with the clinical diagnosis criteria of branchio-otic syndrome.A no-vel heterozygous variant c.963dupT(p.E322X)in EYA1 gene was identified,which co-segregated with the branchi-o-otic syndrome phenotype in the family.The variant was a nonsense variant resulting in the premature appearance of the stop codon.According to the American College of Medical Genetics and Genomics(ACMG)guidelines and the criteria,the variant was classified as pathogenic.Conclusion We identified a novel pathogenic variant EYA1:c.963dupT(p.E322X)in a family with branchio-otic syndrome.