OBJECTIVETo explore the potential molecular mechanisms for Bushen Tiaojing Recipe (BTR) improving the endocrine function of ovarian granular cells by observing the effect of BTR containing serum on follicle stimulating hormone/cyclic adenosine monophosphate-protein kinase A (FSH/ cAMP-PKA) pathway in in vitro cultured human ovarian granular cells.

METHODSThe primary ovarian granular cells collected from in vitro fertilization-embryo transfer patients were cultured for 24 h. The human and rat serum containing different concentrations of BTR (low, medium, high dose), and their normal serums were co-incubated with ovarian granular cells for 48 h respectively, and then they were divided into the low, medium, high dose BTR groups and the control group. The levels of estradiol (E2), progesterone (P), and cyclic adenosine monophosphate (cAMP) in the culture medium were measured by radioimmunoassay. The protein expression of FSHR in ovarian granular cells was detected by Western Blot. The mRNA expression of follicle stimulating hormone receptor (FSHR) and P450 aromatase (P450arom) in ovarian granular cells were detected by Real-time PCR.

RESULTSIn human BTR containing serum groups: Compared with control group, the levels of E2 and cAMP in the culture medium were higher (both P < 0.05) in the medium and high dose BTR groups; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells increased (all P < 0.01), the mRNA expressions of P450arom in ovarian granular cells were higher (P < 0.05, P< 0.01) in the medium and high dose BTR groups. In rat BTR containing serum groups: Compared with the control group, the levels of E2 in the culture medium were higher (all P < 0.01), cAMP in the culture medium were higher (P < 0.05, P < 0.01) in the medium and high dose BTR group; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells were higher (all P < 0.01), the mRNA expression of P450arom in ovarian granular cells increased in the medium and high dose BTR groups (P < 0.05, P < 0.01).

CONCLUSIONBTR could possibly improve the endocrine function of ovarian granular cells by regulating main effector molecules FSHR, cAMP, P450arom, and E2 in FSH/cAMP-PKA pathway of ovarian granular cells.

Cells, Cultured ; Cyclic AMP-Dependent Protein Kinase Type I ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Follicle Stimulating Hormone ; metabolism ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Humans ; Serum ; chemistry ; Signal Transduction ; drug effects

OBJECTIVETo study the effects of huai qi huang, a traditional Chinese medicine, on cytokines Th1, Th2 and Th17 levels and alveolar macrophage phagocytosis in asthmatic rats sensitized by ovalbumin (OVA).

METHODSForty male Sprague-Dawley rats were randomly divided into five groups: normal control, untreated asthma, budesonide-treated, huai qi huang-treated and budesonide+huai qi huang-treated asthma (n=8 each). Asthma was induced by OVA sensitization and challenge. The levels of IL-4, IFN-γ and IL-17 in plasma and bronchoalveolar lavage fluid (BALF) were measured using ELISA. The phagocytosis of alveolar macrophages which were isolated and purified from BALF was evaluated by the colorimetric assay.

RESULTSThe levels of IL-4 and IL-17 increased, in contrast, the IFN-γ level decreased in plasma and BALF in the untreated asthma group compared with those in the normal control group. The IFN-γ level in the huai qi huang-treated asthma group was higher than that in the untreated asthma group. The IFN-γ level increased and the IL-17 level decreased more significantly in the budesonide+huai qi huang-treated asthma group when compared with the budesonide and huai qi huang alone treatment groups. The phagocytosis of alveolar macrophages in the untreated asthma group was lower than that in the normal control group. Huai qi huang alone or combined with budesonide increased the phagocytosis of alveolar macrophages compared with the normal control, untreated asthma and budesonid-treated asthma groups. The levels of IFN-γ in plasma and BALF were positively correlated with the phagocytosis of alveolar macrophages.

CONCLUSIONSThe levels of IL-4 and IL-17 increase and the IFN-γ level decreases in plasma and BALF, and the phagocytosis of alveolar macrophages decreases in asthmatic rats. Huai qi huang treatment may increase the IFN-γ expression in plasma and BALF and the phagocytosis of alveolar macrophages in asthmatic rats. There is a synergistic effect between huai qi huang and glucocorticoids.

Animals ; Asthma ; drug therapy ; immunology ; Cytokines ; biosynthesis ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; Medicine, Chinese Traditional ; Phagocytosis ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes, Helper-Inducer ; immunology ; Th1 Cells ; immunology ; Th17 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo investigate the urinary neutrophil gelatinase-associated lipocalin (NGAL) concentration in children with idiopathic nephrotic syndrome (INS) and its clinical significance.

METHODSThirty-four children newly diagnosed with INS received oral prednisone for 4 weeks. Patients whose urinary protein did not become negative were classified as steroid-resistant nephrotic syndrome (SRNS) group, while those whose urinary protein did become negative were classified as steroid-sensitive nephrotic syndrome (SSNS) group. Morning midstream urine specimens were collected from all patients before use of prednisone and after 1, 2, 3, and 4 weeks of treatment with prednisone. Enzyme-linked immunosorbent assay was used to measure the urinary NGAL concentration. Meanwhile, urinary creatinine (Cr) concentration was measured, and urinary NGAL concentration in a single urine collection was adjusted according to the urinary Cr excretion. The two groups were compared in terms of urinary NGAL/Cr ratio.

RESULTSCompared with the SRNS group, the SSNS group had significantly decreased urinary NGAL/Cr ratios after 3 and 4 weeks of prednisone treatment (P < 0.05). Compared with the SRNS group, the SSNS group had a significantly decreased urinary β2-MG/Cr ratio after 4 weeks of prednisone treatment (P < 0.05). In both groups, urinary NGAL/Cr ratio was positively correlated with urinary protein/Cr ratio (r = 0.510, P < 0.01). The results of ROC curve analysis showed when diagnostic cut-off point of urinary NGAL/Cr was 0.043 by 3 weeks after treatment, sensitivity and specificity achieved 100% and 79.2% respectively.

CONCLUSIONSUrinary NGAL/Cr ratio remains high in children with SRNS, while this ratio decreases gradually during prednisone treatment in children with SSNS, and it falls ahead of urinary β2-MG/Cr ratio. These results suggest that dynamic monitoring of urinary NGAL/Cr ratio is useful for early judgment of response to prednisone in patients with INS.

Acute-Phase Proteins ; urine ; Child ; Child, Preschool ; Creatinine ; urine ; Female ; Humans ; Lipocalin-2 ; Lipocalins ; urine ; Male ; Nephrotic Syndrome ; drug therapy ; urine ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins ; urine ; beta 2-Microglobulin ; urine

OBJECTIVETo examine the transfection of Homeobox A13 (HOXA13) on epithelial-mesenchymal transition (EMT) and the expression of bone morphogenetic protein-7 (BMP-7) induced by albumin-overload in human kidney tubular epithelial cells (HKCs).

METHODSThe cultured HKCs were treated with 20 mg/mL human serum albumin (HSA) for 48 hours. Protein expression of cytokeratin (CK), vimentin and HOXA13 in the HKCs was assessed by Western blot. Protein expression of CK, vimentin, and BMP-7 was also detected in HKCs transfected with lipofectamine contained HOXA13 DNA.

RESULTSHSA induced EMT in HKCs, presented by decreased CK expression (P<0.01) and increased vimentin expression (P<0.01). The up-regulated expression of HOXA13 transfected by lipofectamine inhibited the level of EMT induced by HSA in HKCs (P<0.05). The decreased rate of BMP-7 protein expression induced by HSA was inhibited by over-expressed HOXA13 in HKCs (P<0.05).

CONCLUSIONSTransfection of HOXA13 in HKCs could inhibit the degree of EMT induced by albumin-overload, possibly by increasing BMP-7 expression.

Bone Morphogenetic Protein 7 ; genetics ; Cells, Cultured ; Epithelial Cells ; metabolism ; Epithelial-Mesenchymal Transition ; Homeodomain Proteins ; physiology ; Humans ; Keratins ; genetics ; Kidney Tubules ; metabolism ; Transfection ; Vimentin ; genetics

BACKGROUNDHIV transmission between discordant couples has become an important source of new infections in China. To describe the seroconversion rate among serodiscordant couples and to identify salient behavioral and clinical risk factors including ART that affect heterosexual HIV transmission risk among couples in rural China.

METHODSLongitudinal follow-up of an open cohort of HIV serodiscordant couples took place between 2007 and 2011 in Zhumadian, a city in southern Henan province in China, where blood plasma selling in 1990s led to a regional HIV epidemic. Annual follow-up included separate face-to-face interviews of husbands and wives, and HIV antibody testing for non-index partners. Cox proportional-hazard modeling was used to assess the relationship between HIV seroconversion and covariates of interest.

RESULTSBy the end of 2011, 4499 HIV serodiscordant couples had been enrolled in at least two follow-up interviews; 100 non-index partners seroconverted during the entire observation period for an incidence rate of 0.82 per 100 person-years (95% CI: 0.66-0.99). The incidence rates by the end of 2008, 2009, and 2010 were 2.14, 1.51, and 0.90 per 100 personyears respectively. Always using condoms in the past year of sex, gender of the index partner, frequency of sex, and ART exposure were all significant predictors of HIV seroconversion in the negative spouse. ART was highly protective against seroconversion whether the index partner was actively receiving treatment at the last follow-up (RR = 0.05, 95% CI, 0.01-0.16) or if the index partner had ever received ART (RR = 0.01, 95% CI, 0.00, 0.12). The risk of seroconversion in the nonindex spouse also decreased the longer the duration of the index partner's exposure to ART.

CONCLUSIONSART exposure and always using condom were highly protective against HIV seroconversion in the negative spouse. HIV incidence in serodiscordant couples has been decreasing over time, associated with ART treatment time within 7 years in the index partner. Gender of the index spouse and frequency of sex were also important predictors. Treatment as part of a combination prevention package may be a feasible method of HIV control in this population.

Adult ; Aged ; Anti-Retroviral Agents ; therapeutic use ; China ; epidemiology ; Cohort Studies ; Female ; HIV Infections ; drug therapy ; transmission ; HIV Seropositivity ; Heterosexuality ; Humans ; Male ; Middle Aged ; Proportional Hazards Models ; Risk Factors

OBJECTIVE: To evaluate the efficacy and safety of tacrolimus in the treatment of children with myasthenia gravis (MG). METHODS: A total of 28 children with MG were treated with tacrolimus. MG-Activities of Daily Living (MG-ADL) scale was used to assess clinical outcome and safety after 1, 3, 6, 9, and 12 months of treatment. RESULTS: After tacrolimus treatment, the MG-ADL score at 1, 3, 6, 9 and 12 months was lower than that at baseline (P<0.05), and the MG-ADL score showed a gradually decreasing trend. The response rates to tacrolimus treatment at 1, 3, 6, 9, and 12 months were 59%, 81%, 84%, 88%, and 88% respectively. At 6, 9, 12, and 18 months of treatment, 4, 13, 14, and 15 children respectively were withdrawn from prednisone. No recurrence was observed during treatment. Major adverse reactions/events were asymptomatic reduction in blood magnesium in 5 children and positive urine occult blood in 1 child, which turned negative without special treatment, and tacrolimus was not stopped due to such adverse reactions/events. One child was withdrawn from tacrolimus due to recurrent vomiting. According to CYP3A5 genotypes, all of the patients were divided into two groups: slow metabolic type (n=19) and non-slow metabolic type (fast metabolic type + intermediate type; n=9). The non-slow metabolism group received a higher dose of tacrolimus, but had a lower trough concentration of tacrolimus than the slow metabolism group (P<0.05). The slow metabolism group had a higher response rates to tacrolimus treatment than the non-slow metabolism group (P<0.05). CONCLUSIONS Tacrolimus appears to be effective and safe in the treatment of children with MG and is thus an option for immunosuppressive therapy. CYP3A5 genotyping has a certain guiding significance for determining the dosage of tacrolimus.
Activities of Daily Living ; Child ; Humans ; Immunosuppressive Agents ; Myasthenia Gravis ; drug therapy ; Neoplasm Recurrence, Local ; Tacrolimus ; therapeutic use

Objective To study the clinical characteristics of children with anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis(AAV).Methods A retrospective analysis was conducted on the clinical data of 25 children diagnosed with AAV at the Second Xiangya Hospital of Central South University from January 2010 to June 2022.Results Among the AAV children,there were 5 males and 20 females,with a median age of onset of 11.0 years.Involvement of the urinary system was seen in 18 cases(72%);respiratory system involvement in 10 cases(40%);skin involvement in 6 cases(24%);eye,ear,and nose involvement in 5 cases(20%);joint involvement in 4 cases(16%);digestive system involvement in 2 cases(8%).Eleven cases underwent kidney biopsy,with 5 cases(46%)showing focal type,2 cases(18%)showing crescentic type,2 cases(18%)showing mixed type,and 2 cases(18%)showing sclerotic type.Immune complex deposits were present in 5 cases(45%).Seven cases reached chronic kidney disease(CKD)stage Ⅴ,with 2 cases resulting in death.Two cases underwent kidney transplantation.At the end of the follow-up period,2 cases were at CKD stage Ⅱ,and 1 case was at CKD stage Ⅲ.Of the 16 cases of microscopic polyangiitis(MPA)group,13(81%)involved the urinary system.Of the 9 cases of granulomatosis with polyangiitis(GPA),6 cases(66%)had sinusitis.Serum creatinine and uric acid levels were higher in the MPA group than in the GPA group(P<0.05),while red blood cell count and glomerular filtration rate were lower in the MPA group(P<0.05).Conclusions AAV is more common in school-age female children,with MPA being the most common clinical subtype.The onset of AAV in children is mainly characterized by renal involvement,followed by respiratory system involvement.The renal pathology often presents as focal type with possible immune complex deposits.Children with MPA often have renal involvement,while those with GPA commonly have sinusitis.The prognosis of children with AAV is poor,often accompanied by renal insufficiency.

Objective To study the correlation of anti-C1q antibodies with active systemic lupus erythematosus(SLE)and lupus nephritis(LN)in children,as well as their diagnostic value for active SLE and LN.Methods A retrospective selection of 90 hospitalized children with SLE at the Children's Medical Center of Second Xiangya Hospital,Central South University from January 2016 to March 2019 as the SLE group,all of whom were tested for anti-C1q antibodies.A control group was formed by collecting 70 hospitalized children with other autoimmune diseases(OAD)during the same period.The differences in anti-C1q antibody levels were compared between two groups.The correlation of anti-C1q antibodies with various indicators of SLE and LN was analyzed,and the diagnostic value of anti-C1q in SLE and LN was evaluated.Results The serum levels of anti-C1q antibodies in the SLE group were higher than those in the OAD group(P<0.05).The SLE disease activity index score was positively correlated with anti-C1q antibodies(rs=0.371,P<0.001)and positively correlated with anti-double-stranded DNA antibodies(rs=0.370,P<0.001).The sensitivity and specificity of anti-C1q antibodies for diagnosing active SLE were 89.90%and 53.90%,respectively,with an area under the curve of 0.720(P<0.05)and a critical value of 5.45 U/mL.The sensitivity and specificity of anti-C1q antibody levels for diagnosing active LN were 58.50%and 85.00%,respectively,with an area under the curve of 0.675(P<0.05)and a critical value of 22.05 U/mL.Conclusions Anti-C1q antibodies can serve as non-invasive biomarkers for evaluating the activity of SLE or predicting the activity of LN in children.

Objective: To investigate the clinical and biological characteristics of familial platelet disorder (FPD) with germline Runt-related transcription factor (RUNX) 1 mutations. Methods: Patients diagnosed with myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML) with RUNX1 mutations from February 2016 to December 2021 in Wuhan No.1 Hospital underwent pedigree analysis and were screened for gene mutations (somatic and germline). Patients diagnosed with FPD with germline RUNX1 mutations were enrolled and evaluated in terms of clinical characteristics and biological evolution. Bioinformatics analysis was used to assess the pathogenicity of mutations and to analyze the effect of mutated genes on the function of the corresponding protein. Results: Germline RUNX1 mutations were detected in three out of 34 patients suffering from MDS/AML who had RUNX1 mutations. A pedigree of FPD with RUNX1 (RUNX1-FPD) c.562A>C and RUNX1 c.1415T>C mutations was diagnosed, and the mutations were of patrilineal origin. Bioinformatics analysis indicated that the locus at positions 188 and 472 in the AML-1G type of RUNX1 was highly conserved across different species, and that variations might influence functions of the proteins. The mutations were evaluated to be highly pathogenic. Of the nine cases with germline RUNX1 mutations: two patients died due AML progression; one case with AML survived without leukemia after transplantation of hemopoietic stem cells; four patients showed mild-to-moderate thrombocytopenia; two cases had no thrombocytopenia. During the disease course of the proband and her son, mutations in RUNX1, NRAS and/or CEBPA and KIT appeared in succession, and expression of cluster of differentiation-7 on tumor cells was enhanced gradually. None of the gene mutations correlated with the tumor were detected in the four cases not suffering from MDS/AML, and they survived until the end of follow-up. Conclusions: RUNX1-FPD was rare. The mutations c.562A>C and c.1415T>C of RUNX1 could be the disease-causing genes for the family with RUNX1-FPD, and these mutations could promote malignant transformation. Biological monitoring should be carried out regularly to aid early intervention for family members with RUNX1-FPD.
Humans ; Female ; Germ-Line Mutation ; Core Binding Factor Alpha 2 Subunit/genetics* ; Pedigree ; Blood Platelet Disorders/complications* ; Leukemia, Myeloid, Acute/genetics*

Objective To evaluate the survival and prognostic factors of esophageal cancer treated with definitive ( chemo ) radiotherapy by applying novel radiation techniques including three-dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT). Methods Clinical data of 2762 patients with non-operated esophageal squamous cell carcinoma who underwent definitive ( chemo ) radiotherapy from 2002 to 2016 in 10 hospitals were retrospectively analyzed.The prognostic factors were also identified and analyzed. Results The median follow-up time was 60. 8 months. The 1-, 2-, 3-and 5-year overall survival (OS) of all patients was 71. 4％,48. 9％,39. 3％,and 30. 9％,respectively.The 1-,2-,3-and 5-year progression-free survival (PFS) was 59.5％,41.5％,35.2％,and 30％,respectively.The median survival was 23 months.The median time to progression was 17. 2 months.Multivariate analysis demonstrated that age, primary tumor location, clinical stage, tumor target volume, EQD2 and treatment mode were the independent prognostic factors for OS.Primary tumor location,clinical stage,tumor target volume and EQD2 were the independent prognostic factors for PFS. Conclusions In this first large-scale multi-center retrospective analysis of definitive ( chemo) radiotherapy for esophageal squamous cell carcinoma in China, the 5-year OS of patients with esophageal squamous cell carcinoma is significantly improved by 3DCRT, IMRT combined with chemotherapy drugs. However, the findings remain to be validated by prospective clinical trials with high-level medical evidence.