This study aims to explore the mechanism through which Yishen Jiangtang Decoction(YSJTD) regulates endoplasmic reticulum stress(ERS)-mediated NOD-like receptor thermal protein domain associated protein 3(NLRP3) inflammasome to improve diabetic nephropathy(DN) in db/db mice. Thirty db/db mice were randomly divided into the model group, YSJTD group, ERS inhibitor 4-phenylbutyric acid(4-PBA) group, with 10 mice in each group. Additionally, 10 db/m mice were selected as the control group. The YSJTD group was orally administered YSJTD at a dose of 0.01 mL·g~(-1), the 4-PBA group was orally administered 4-PBA at a dose of 0.5 mg·g~(-1), and the control and model groups were given an equal volume of carboxylmethyl cellulose sodium. The treatments were administered once daily for 8 weeks. Food intake, water consumption, and body weight were recorded every 2 weeks. After the intervention, fasting blood glucose(FBG), glycosylated hemoglobin(HbA1c), urine microalbumin(U-mALB), 24-hour urine volume, serum creatinine(Scr), and blood urea nitrogen(BUN) were measured. Inflammatory markers interleukin-1β(IL-1β) and interleukin-18(IL-18) were detected using the enzyme-linked immunosorbent assay(ELISA). Renal pathology was assessed through hematoxylin-eosin(HE), periodic acid-Schiff(PAS), and Masson staining, and transmission electron microscopy(TEM). Western blot was used to detect the expression levels of glucose-regulated protein 78(GRP78), C/EBP homologous protein(CHOP), NLRP3, apoptosis-associated speck-like protein containing CARD(ASC), cysteinyl aspartate-specific proteinase(caspase-1), and gasdermin D(GSDMD) in kidney tissues. The results showed that compared to the control group, the model group exhibited poor general condition, increased weight and food and water intake, and significantly higher levels of FBG, HbA1c, U-mALB, kidney index, 24-hour urine volume, IL-1β, and IL-18. Compared to the model group, the YSJTD and 4-PBA groups showed improved general condition, increased body weight, decreased food intake, and lower levels of FBG, U-mALB, kidney index, 24-hour urine volume, and IL-1β. Specifically, the YSJTD group showed a significant reduction in IL-18 levels compared to the model group, while the 4-PBA group exhibited decreased water intake and HbA1c levels compared to the model group. Although there was a decreasing trend in water intake and HbA1c in the YSJTD group, the differences were not statistically significant. No significant differences were observed in BUN, Scr, and kidney weight among the groups. Renal pathology revealed that the model group exhibited more severe renal damage compared to the control group. Kidney sections from the model group showed diffuse mesangial proliferation in the glomeruli, tubular edema, tubular dilation, significant inflammatory cell infiltration in the interstitium, and increased glycogen staining and blue collagen deposition in the basement membrane. In contrast, the YSJTD and 4-PBA groups showed varying degrees of improvement in renal damage, glycogen staining, and collagen deposition, with the YSJTD group showing more significant improvements. TEM analysis indicated that the model group had extensive cytoplasmic edema, homogeneous thickening of the basement membrane, fewer foot processes, and widening of fused foot processes. In the YSJTD and 4-PBA groups, cytoplasmic swelling of renal tissues was reduced, the basement membrane remained intact and uniform, and foot process fusion improved.Western blot results indicated that compared to the control group, the model group showed upregulation of GRP78, CHOP, GSDMD, NLRP3, ASC, and caspase-1 expression. In contrast, both the YSJTD and 4-PBA groups showed downregulation of these markers compared to the model group. These findings suggest that YSJTD exerts a protective effect against DN by alleviating NLRP3 inflammasome activation through the inhibition of ERS, thereby improving the inflammatory response in db/db DN mice.
Animals ; Endoplasmic Reticulum Stress/drug effects* ; Diabetic Nephropathies/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Inflammasomes/drug effects* ; Male ; Kidney/pathology* ; Endoplasmic Reticulum Chaperone BiP ; Humans ; Interleukin-18/genetics* ; Mice, Inbred C57BL

Objective: To report the clinical manifestations and laboratory features of five patients with congenital thrombotic thrombocytopenic purpura (cTTP) and explore its standardized clinical diagnosis and treatment along with a review of literature. Methods: Clinical data of patients, such as age of onset, disease manifestation, personal history, family history, and misdiagnosed disease, were collected. Treatment outcomes, therapeutic effects of plasma infusion, and organ function evaluation were observed. The relationship among the clinical manifestations, treatment outcomes, and ADAMTS13 gene mutation of patients with cTTP was analyzed. Additionally, detection of ADAMTS13 activity and analysis of ADAMTS13 gene mutation were explored. Results: The age of onset of cTTP was either in childhood or adulthood except in one case, which was at the age of 1. The primary manifestations were obvious thrombocytopenia, anemia, and different degrees of nervous system involvement. Most of the patients were initially suspected of having immune thrombocytopenia. Acute cTTP was induced by pregnancy and infection in two and one case, respectively. ADAMTS13 gene mutation was detected in all cases, and there was an inherent relationship between the mutation site, clinical manifestations, and degree of organ injury. Therapeutic or prophylactic plasma transfusion was effective for treating cTTP. Conclusions: The clinical manifestations of cTTP vary among individuals, resulting in frequent misdiagnosis that delays treatment. ADAMTS13 activity detection in plasma and ADAMTS13 gene mutation analysis are important bases to diagnose cTTP. Prophylactic plasma transfusion is vital to prevent the onset of the disease.
Female ; Pregnancy ; Humans ; Adult ; Blood Component Transfusion ; Plasma ; Purpura, Thrombotic Thrombocytopenic/therapy* ; Mutation ; Purpura, Thrombocytopenic, Idiopathic ; ADAMTS13 Protein/therapeutic use*

We established an indirect ELISA method using Trichinella spiralis trehalase(TsTRE)protein expressed in prokaryotic cells.The TsTRE gene was amplified by RT-PCR and ligated into the pCold I plasmid,which was expressed in E.coli BL21 competent cells.The rTsTRE protein was purified through affinity column chromatography.The TsTRE protein was localized with immunofluorescence techniques,and the immunogenicity of rTsTRE was detected by westernblotting.Subse-quently,rTsTRE protein was used as a coating antigen to establish an indirect ELISA.We optimized the antigen-coating con-centration,serum dilution concentration,antigen-coating incubation time,type of blocking solution,blocking incubation time,HRP-labeled goat anti-rabbit IgG serum dilution concentration,HRP-labeled goat anti-rabbit IgG serum incubation time and response time of TMB.Subsequently,the critical value,repeatability,sensitivity,specificity and clinical detection rate of the ELISA were evaluated.Immunofluorescence indicated that trehalase was abundant in the rod-shaped body,tail and epidermis of Trichinella spiralis muscle larvae.Western-blot indicated that rTsTRE protein combined with the positive serum of mice infected with T.spiralis for 42 d;the band was approximately 60 kDa.The established indirect ELISA had a positive threshold of 0.384;the intra-run and inter-run coefficients of variation were 5.504％-7.630％ and 4.664％-9.929％,and did not exceed 10％.The lowest detectable titer was 1:1 280.No cross reaction was observed with antibodies to Clonorchissinensis,Schistosoma ja-ponicum,Ascaris suum,Toxocara gondii and Toxocara canis,and the clinical negative detection rate was 0％.Thus,we suc-cessfully expressed the rTsTRE protein.Moreover,the established indirect ELISA method using the TsTRE protein as the coating antigen had good repeatability,sensitivity,specificity and clinical detectability,and can be applied to the detection of clinical samples.

OBJECTIVE: To explore clinical effects of micro external fixator for the treatment of supracondylar fracture of humerus in children. METHODS: From October 2017 to December 2018, 20 children with supracondylar fracture of humerus (treatment group) were admitted and treated with micro-external fixation after closed reduction, including 14 males and 6 females, aged from 6 to 14 years old with an average of (7.9±2.4) years old, classified to Gartland Ⅲ. Thirty nine children with supracondylar fracture of humerus were admitted as control group from January 2015 to September 2017, and treated with closed reduction Kirschner wire fixation. Among them, including 24 males and 15 females, aged from 6 to 14 years old with an average of (8.1±1.9) years old, classified to GartlandⅢ. Operation time, times of intraoperative fluoroscopy, and complications between two groups were observed and compared, Mayo scoring system at the latest follow up was used to evaluate clinical effect. RESULTS: Twenty children in treatment group were followed up from 6 to 12 months with an average of (8.0±2.5) months, operation time was(30.10±12.50) min, times of intraoperative fluoroscopy was(10.00±2.50). Fifteen patients got excellent results, 3 good, 2 fair according to Mayo elbow joint scoring. No ulnar nerve injury moderate or severe elbow varus occurred in treatment group. Thirty-nine children in control group were followed up from 5 to 13 months with an average of (9.0±3.1) months, operation time was(45.60±18.90) min, times of intraoperative fluoroscopy was(19.00±5.60). Twenty-three patients got excellent results, 12 good, 3 fair and 1 poor according to Mayo elbow joint scoring. One child occurred ulnar nerve injury and recovered at the final follow-up, 1 child occurred severe cubitus varus and 2 children occurred moderate cubitus varus. There was no statistical difference in clinical effect between two groups ( CONCLUSION Treated with closedreduction and mini external fixator fixed on children humerus condyle fracture could receive the same clinical result as closed as reduction gram needle fixation, which has advantages of shorter operation time, less times of intraoperative fluoroscopy, not esay to damage ulnar nerve during operation, less incidence of moderate and severe cubitus varus deformity after operation, and the permeation was simple.
Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; External Fixators ; Female ; Fracture Fixation, Internal ; Humans ; Humeral Fractures/surgery* ; Humerus ; Male

The aim of this paper was to investigate the effect of total saponins from Panax japonicus( SPJ) on cognitive decline of natural aging rats and its mechanism. Thirty male SD rats of eighteen month old were randomly divided into three groups: aged group,10 mg·kg~(-1) SPJ-treated group and 30 mg·kg~(-1) SPJ-treated group. The SPJ-treated groups were given SPJ at the dosages of 10 mg·kg~(-1) and 30 mg·kg~(-1),respectively,from the age of 18 to 24 months. Aged group were lavaged the same amount of saline,10 six-month-old rats were used as control group,with 10 rats in each group. The open field test,novel object recognition and Morris water maze were performed to detect the changes of cognitive function in each group. The changes of synaptic transmission of long-term potentiation( LTP) in hippocampal CA1 region were detected by field potential recording. Western blot was used to detect the protein levels of NLRP3,ASC,caspase-1 and the changes of Glu A1,Glu A2,CAMKⅡ,CREB and phosphorylation of CAMKⅡ,CREB in each group.The results showed that SPJ could improve the decline of cognitive function in aging rats,reduce the damage of LTP in the hippocampal CA1 region of aged rats,and decrease the expression of NLRP3,ASC,caspase-1 in aging rats. At the same time,SPJ could enhance the membrane expression of AMPA receptor( Glu A1 and Glu A2),and increase the expression of p-CAMKⅡand p-CREB in aging rats.SPJ could improve cognitive decline of natural aging rats,and its mechanism may be related to regulating NLRP3 inflammasome,thus regulating the membrane expression of AMPA receptor,and enhancing the expression phosphorylation of CAMKⅡ and CREB.
Aging ; Animals ; CA1 Region, Hippocampal ; physiology ; Cognition ; drug effects ; Inflammasomes ; metabolism ; Long-Term Potentiation ; Male ; NLR Family, Pyrin Domain-Containing 3 Protein ; metabolism ; Panax ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology

OBJECTIVETo investigate the prevalence, current treatment, and clinical characteristics of asthma, as well as the risk factors for this disease, among children aged 0-14 years in 2010 in urban Zhongshan, China.

METHODSA total of 10 336 children aged 0-14 years were selected from urban Zhongshan by cluster random sampling. The Third National Childhood Asthma Epidemiological Questionnaire 2010 was used to analyze the prevalence, current treatment, and clinical characteristics of childhood asthma, as well as the risk factors for this disease.

RESULTSAsthma was diagnosed in 179 cases (1.73%). The prevalence of asthma in male children was significantly higher than that in female children (2.25% vs 1.16%; P<0.01). Of the 179 patients, severe attacks were common in 104 cases (58.1%), 110 cases (61.5%) had slow onset, 102 cases (57.0%) had gradually relieved conditions, 61 cases (34.1%) suffered from asthma during seasonal transition, and 150 cases (83.8%) developed asthma due to respiratory tract infection. Among all asthmatic children, 71.5% had been treated with inhaled corticosteroids, and 71.5% had been treated with bronchodilator. The multivariate logistic regression analysis showed that a history of penicillin allergy, a family history of allergy, food allergy, eczema, allergic rhinitis, cesarean delivery, family mould, and perinatal passive smoking were independent risk factors for childhood asthma.

CONCLUSIONSThe prevalence of childhood asthma in urban Zhongshan is on a high level, and is associated with gender. The treatment of asthma has been standardized, but still needs further improvement. The onset of asthma attack is influenced by various factors.

Adolescent ; Asthma ; epidemiology ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Risk Factors ; Seasons ; Time Factors

Naringenin (1) was transformed to three metabolites (2-4) by Mucor sp. Based on LCMS(n)-IT-TOF and NMR spectroscopic data, 2-4 were identified as naringenin-7-O-sulphate, naringenin-4'-O-sulphate, and naringenin-5-O-sulphate, respectively. These results might provide hints to the mammalian/human metabolism of naringenin.
Biotransformation ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Flavanones ; chemistry ; metabolism ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Mucor ; metabolism ; Sulfates ; metabolism

OBJECTIVETo investigate the incidence of wheezing and chronic cough in children aged 3-14 years in different living areas of Zhongshan, China.

METHODSAccording to the unified program of the 2010 national epidemiological survey of asthmatic children aged 0-14 years, a questionnaire survey of the children aged 3-14 years in rural and urban areas of Zhongshan was conducted.

RESULTSA total of 15 763 children were included in the survey. Among all participants, 8 248 were from the urban area, and 7 515 from the rual area; 8 306 were boys, and 7 457 were girls. The percentage of children with a history of wheezing was significantly higher in the urban group than in the rural group (6.6% vs 3.2%; P<0.05), and it was significantly higher in boys than in girls for each group (P<0.05). The urban group had a significantly higher percentage of individuals who had wheezing in the past one year than the rural group (2.8% vs 1.5%; P<0.05), and this percentage was significantly higher in boys than in girls for each group (P<0.05). Compared with the rural group, the urban group had a significantly higher percentage of individuals who had chronic cough (duration 1 year) (7.9% vs 3.1%; P<0.05). The above indices were relatively high in children under 6 years of age, and all but the percentage of girls who had wheezing in the past one year in both rural and urban areas showed significant differences between all age groups.

CONCLUSIONSThe incidence of wheezing and chronic cough varies with age, sex, and living area for children aged 3-14 years in Zhongshan, and it is relatively high under 6 years of age, in the urban area, and among boys.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Chronic Disease ; Cough ; epidemiology ; Female ; Humans ; Incidence ; Male ; Respiratory Sounds ; Surveys and Questionnaires

OBJECTIVETo investigate sperm DNA integrity in male infertility patients with hepatitis B virus (HBV) infection.

METHODSThis study included 90 infertile men with HBV infection (group A), 82 infertile men without HBV infection (group B) and 70 normal fertile men (group C). We detected sperm DNA integrity among the subjects, including DNA fragmentation index (DFI) and high DNA stainability (HDS), by sperm chromatin structure assay (SCSA), and compared them among the three groups.

RESULTSDFI was higher in group A ([28.17 +/- 13.06]%) than in B ([26.64 +/- 9.79]%) and C ([15.67 +/- 4.73]%), significantly higher in A and B than in C (P < 0.05) but with no significant difference between A and B (P > 0.05). HDS was higher in group A ([10.83 +/- 5.601]%) than in B ([9.04 +/- 3.48]%) and C ([8.04-2.25]%), with significant difference between A and C (P < 0.05).

CONCLUSIONSperm DNA integrity of infertile males is significantly different from that of normal fertile men, and infertility with HBV infection further impairs sperm DNA, which is manifested by abnormal sperm nuclear maturity.

Adult ; Case-Control Studies ; Chromatin ; DNA ; genetics ; DNA Damage ; Hepatitis B ; pathology ; Hepatitis B virus ; Humans ; Infertility, Male ; genetics ; virology ; Male ; Sperm Count ; Spermatozoa ; pathology ; Young Adult

This study was purposed to investigate the apoptosis-inducing effect of Annexin A2 gene (AnxA2) on multiple myeloma (MM) cells and its mechanisms. The human MM cell lines U266 and RPMI8226 were transfected by using siRNA targeting at AnxA2; the expressions of AnxA2 mRNA and protein in the siRNA-transfected cells were detected by real-time PCR and Western blot, respectively; the cell apoptosis was assayed by flow cytometry. The results showed that silencing AnxA2 gene by siRNA resulted in decreased expressions of AnxA2 gene and protein, increased apoptosis of U266 and RPMI8226 cell lines (P < 0.05), at the same time resulted in down-regulation of apoptosis-related gene expressions including p65NF-κB, IL-2, IL-6 (P < 0.05), and up-regulation of P53 gene expression (P < 0.05). It is concluded that the AnxA2 silence plays a promoting role in apoptosis of MM cell lines U266 and RPMI8226.
Annexin A2 ; genetics ; Apoptosis ; genetics ; Cell Line, Tumor ; Humans ; Multiple Myeloma ; genetics ; pathology ; RNA, Small Interfering ; genetics