OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

Objective To observe the combined poisonous effects of fluoride and aluminum on sex hormone of male rats.Methods Sixteen weaned SD healthy male rats aged two week were selected and divided into control group,aluminum group,fluoride group,fluorine-aluminum group,four rats in each group.All rats in the experimental groups were fed with corn collected from the prevailng areas containing different fluorine contents respectively for 90 days.Serum testosterone(T)and estradiol(E2)were detected.Results Compared separatelv with the control group[(3.317±0.635)μg/L],serum T level of fluorine-aluminum group[(15.994±6.558)μg/L]was higher(P＜0.05),but aluminum[(8.134±3.134)μg/L]and fluorine[(1.868±0.367)μg/L]groups had no significant differences(P＞0.05).Compared separately with the control group[(0.319±0.072)nmol/L],E2 level of the fluorine group[(0.172±0.030)nmol/L]being lower(P＜0.05),and it was not significant differences(P＞0.05)in the control group when compared with aluminum group[(0.282±0.012)nmol/L],and fluorine-aluminum group[(0.265±0.047)nmol/L].Fluorine and aluminum interacted with each other(F=9.82,P＜0.05).Conclusion The combined poisonous effects of fluorine and aluminum may influence sex hormone levels of male rats.

Objective:To evaluate the practical application of intra-abdominal pressure (IAP) monitoring product in patients receiving enteral nutrition (EN) and with high risk of intra-abdominal hypertension/abdominal compartment syndrome (IAH/ACS).Methods:Patients receiving EN treatment from Changzheng hospital were randomly divided as experimental group (measuring IAP with pressure monitoring product,n =60) and control group (measuring IAP with conventional method,n =60).The clinical data of gastrointestinal complications,gastric residual volume,IAP and completion of target infusions were collected and analyzed.Results:The incidence of gastrointestinal complications in experimental group patients significantly decreased comparing with the control group patients (7.92％ vs 28.33％,P ＜ 0.01).The levels of gastric residual volume and IAP in experimental group patients were lower than those in control group patients[(50.12 ± 10.66) ml vs.(101.54 ± 25.81) ml,(7.17 ± 1.84) cmH2O vs (12.36 ± 2.51) cmH2O,P ＜0.05].Moreover,The experimental group patients had a shorter period to achieve target infusions and higher proportion of completion treatments [88.3％ vs 71.7％,(2.94 ± 0.78)d vs (3.78 ± 1.02)d,P ＜ 0.05)].Conclusion:As utilizing pressure monitoring products to assist EN treatment for patients with high risk of IAH/ACS could achieve lower incidence of gastrointestinal complications,excellent EN tolerance,and improve target feeding,its clinical application should be extended.

Abstract: Objective　To investigate the clinical and laboratory characteristics of pulmonary infection caused by Nocardia otitidiscaviarum. Methods　The clinical data of a patient with pulmonary infection caused by Nocardia otitidiscaviarum were reported, and the clinical characteristics, laboratory characteristics and drug sensitivity of pulmonary infection caused by Nocardia otitidiscaviarum were summarized in combination with the relevant literature at home and abroad from January 2010 to December 2022. Results　A 67-year-old female patient was admitted to the hospital on June 30, 2020 because of "repeated chest tightness and shortness of breath for 3 years, aggravated cough, expectoration and fever". The sputum, alveolar lavage fluid and blood of the patient were collected for culture, and the detected pathogenic bacteria were identified. There are pathogenic bacteria growing in sputum and alveolar lavage fluid, which are identified as Nocardia otitidiscaviarum by Autof ms mass spectrometer. According to the results of pathogenic bacteria and the patient's condition, meropenem combined with compound sulfamethoxazole tablets were given anti-infection treatment, and the patient's condition improved and discharged. Conclusion　The clinical manifestations and imaging features of nocardiosis are lack of specificity, and are prone to misdiagnosis and missed diagnosis. Etiology is the key to disease diagnosis, and clinical examination and culture should be conducted in time.

Abstract: Objective　To analyze the emergency response and long-term intervention effects after the detection of infectious snails epidemic by loop-mediated isothermal amplification (LAMP) assays in Hannan District, Wuhan City, and to explore the application of LAMP in early surveillance and early-warning of schistosomiasis transmission. Methods Snails picked up by the risk monitoring system in Hannan District were examined by anatomical microscopy and LAMP technology to identify the schistosomiasis infection. Emergency response and intensive intervention were initiated in the environment where positive snails appeared, and the long-term effects were evaluated. Results In May 2018, the infectious snails were detected by LAMP technology in Hannan District, and the positive snails were located in Zhujiacha, Dongzhuang Village, Obstacles and weeds were removed and buried by machine in Zhujiacha. 12 700 m2 of snails were killed by drugs, and the mortality rate of snails was more than 80%; no new seropositive persons were found in the emergency examination within 500 m of the positive snail sites. 506 people were examined in Dong Zhuang Village at the end of the year, and 30 positive IHA cases were detected with a blood positive rate of 5.93%, no positive fecal test was found, and all positive blood test patients took preventive medication. The monitoring results of sentinel rats and wild feces were all negative. Health education was carried out, 7 warning signs were deployed and refreshed, and 500 publicity brochures were distributed. After nearly three years of intensified intervention and monitoring in the villages where the positive environment is located, and the density of snails on the stubborn snail has dropped from 0.094/frame to 0.027/frame, and the positive rate of blood test in Dongzhuang Village has steadily dropped from 5.93% to 3.74%. Conclusions The infected snails missed by microscopy were detected by LAMP in Hannan District, which created conditions for the rapid emergency treatment of environment and elimination of positive snail and improved the sensitivity of the surveillance and early warning system in transmission-interrupted areas.

Objective To compare the differences in virulence-related factor aspartate protease,biofilm formation,and gene expression among clinical isolates of Candida parapsilosis(C.parapsilosis).Methods Gene sequencing and microsatellite typing(MT)method were adopted to identify C.parapsilosis isolated from patients with clinical fungal infection.The production of secreted aspartate protease and biofilm formation ability of each strain were de-tected,and the expression of biofilm formation related-genes BCR1,EFG1,and HWP1,as well as aspartate prote-ase virulence genes SAPP1,SAPP2,SAPP3 were compared among the strains.Results A total of 8 clinically iso-lated C.parapsilosis strains were collected,all of which were identified as genotype Ⅰ.Based on microsatellite ty-ping results,8 clinical strains were divided into 4 microsatellite types.G1,G2,and G3 strains isolated from the urine,peripherally inserted central catheters(PICC),and blood of patient A were of different subtypes.J1,J2,J3,J4,and J5 strains were of the same type,and isolated from blood specimens of patient B at different periods.All 8 clinical strains could form biofilm,and their biofilm formation ability was higher than that of the standard strain of C.parapsilosis(ATCC 22019).G1,G3 and J5 strains had strong biofilm formation ability,J1,J2,J3,and J4 strains had moderate biofilm formation ability,and G2 strain had weak biofilm formation ability.All of the eight clinical isolates secreted aspartate protease,and their in vitro expression levels of the enzyme were higher than that of the standard strain(ATCC 22019).G3,G1,and G2 strains showed low,moderate,and high in vitro enzyme expression respectively,with statistical differences(all P＜0.05).Enzyme expressed moderately in J1 and J5 strains,and highly in J2,J3,and J4 strains.Difference between moderate and high expressions was statistically significant(P＜0.05).The expression levels of biofilm formation genes BCR1,EFG1,and HWP1 in various strains isolated from patients A and B increased.In strains isolated from patient A,the expression level of EFG1 gene in G1 strain was higher than that in G2 strain(P＜0.05).There was no statistically significant difference in BCR1,EFG1,and HWP1 gene expression levels among strains isolated from patient B.The expression levels of as-partate protein genes(SAPP1,SAPP2,and SAPP3)in various strains isolated from patients A and B increased.The expression levels of SAPP1 and SAPP2 in strain G1 were higher than those in G2 and G3(both P＜0.05).There was no statistically significant difference in the expression levels of SAPP1,SAPP2,and SAPP3 genes in strains from patient B.Conclusion Clinical isolates of C.parapsilosis have higher biofilm formation and aspartate protease production abilities than standard strain.The expression of virulence factors varies among strains isolated from different specimens,while there is no significant difference in the expression of virulence factors among strains isolated at different periods.Patients may have been infected with different MT types of C.parapsilosis in multiple sites during the same period.

Aged ; Animals ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Middle Aged ; Orientia tsutsugamushi ; Scrub Typhus ; epidemiology

OBJECTIVETo observe the feasibility of measuring multiple circulating cytokines by protein chips in patients with acute coronary syndrome (ACS).

METHODSCirculating cytokines were determined by protein chips in 50 control cases and 104 ACS cases of unstable angina pectoris (UA, n = 50) and acute myocardial infarction (AMI, n = 54) cases.

RESULTSThe levels of serum MMP-9, sCD40L, CRP, IL-6, H-FABP, cTnI and plasma IL-8, NT-proBNP, sVCAM-1, ET-1 in ACS group were significantly higher than those in the control group (all P < 0.01). Serum H-FABP and cTnI were significantly higher in AMI patients than in UA patients (P < 0.01). Serum MMP-9, sCD40L, CRP, IL-6 and plasma sVCAM-1, ET-1 tended to be higher in AMI patients as compared to UA patients (all P > 0.05). Serum MMP-9, sCD40L, and H-FABP were positively correlated with the severity of angina(r = 0.653, r = 0.745, r = 0.933, P < 0.01, respectively). Serum MMP-9, sCD40L, and H-FABP increased in proportion to the levels of Braunwald angina classes in UA patients (angina class I < angina class II < angina class III, P < 0.01).

CONCLUSIONSCirculating cytokines were significantly increased in ACS patients and positively correlated to Braunwald angina classes in UA patients.

Acute Coronary Syndrome ; blood ; Aged ; Angina Pectoris ; blood ; Angina, Unstable ; blood ; C-Reactive Protein ; metabolism ; CD40 Ligand ; blood ; Cytokines ; blood ; Female ; Humans ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Myocardial Infarction ; blood ; Protein Array Analysis

The study was purposed to investigate the growth inhibitory effect of tanshinones on K562 cell line and the relationship between their structures and cytotoxicity. The modified MTT assay was adopted to measure the inhibitory effect of tanshinones at different concentrations and chemical structures on K562 cells, and the changes of cell morphology were observed by inverted phase contrast microscopy. The results indicated that the tanshinones could inhibit the proliferation of K562 cells effectively, and their cytotoxicities on K562 cells showed concentration- and time-dependent manners. The IC(50) of dihydrotanshinone I, tanshinone I, tanshinone IIA and cryptotanshinone at 24 hours were 0.91, 4.04, 5.95, 13.85 µg/ml at 48 hours were 0.37, 1.35, 1.71, 6.71 µg/ml; at 72 hours were 0.33, 0.46, 0.82, 6.02 µg/ml, respectively. It is concluded that all of the four tanshinones have proliferation inhibitory effect on K562 cell line, among them the dihydrotanshinone I is the most active one, followed by tanshinone I, tanshinone IIA and cryptotanshinone subsequently, indicating that the chemical structure of aromatic ring A of tanshinones can enhance their cytotoxicity and the structure of furan ring C may influence the cytotoxicity, but their mechanism is still remained to be further investigated.
Cell Proliferation ; drug effects ; Diterpenes, Abietane ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; K562 Cells ; Structure-Activity Relationship

The aim of this study was to expand hematopoietic progenitor cells at large scale by magnet stirred culture system. Mononuclear cell from umbilical cord blood were cultured in serum-free medium with stem cell factor, FIT-3 ligand and thrombopoietin. Firstly, the role of magnet on cell growth and colony-forming was studied by static culture on 0, 25 and 50 mT. Then the expansion multiple of cells, colony-forming and expression of surface markers were studied in magnet stirred culture by cell counting, colony-forming assay and flow cytometry. The results indicated that there was no difference in multiple of total cell expansion and numbers of hematopoietic colonies between 0, 25 and 50 mT groups and spinner groups (all p > 0.05). After 7 day cultures, the multiple of total cell expansion in magnet stirred culture was higher than that in static culture (p < 0.01). The numbers of CFU-GM (colony-forming unit-granulocyte/macrophage) and CFU-E (erythroid colony forming unit) in magnet stirred culture were higher than those in static culture, (p < 0.05). The primitive cells (CD34(+), CD34(+)/CD38(-) or CD133(+)) of the expanded cells in magnet stirred culture were less than those in static culture (p < 0.05). However, the CD184(+) or CD62L(+) expanded cells were more than that in static culture (p < 0.05). It is concluded that magnet stirred culture favors the expansion of hematopoietic progenitor cells. The results will be finally confirmed in further in vivo experiments and clinical applications.
Cell Culture Techniques ; methods ; Cell Differentiation ; physiology ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology