Objective To investigate the therapeutic efficacy of artesunate(AS)on polycystic ovary syndrome(PCOS)in mice and explore the potential mechanism primarily.Methods Twenty-five female C57BL/6J mice were randomly divided into Control group,model group(PCOS group),low-and high-dose AS groups(AS15 and AS30 groups)and metformin group(Met group).In addition to the Control group,the mouse model of PCOS was established by subcutaneous injection of dehydroepiandrosterone(DHEA,60 mg/kg)following by a high-fat diet for 21 d.After modeling,AS of 15 and 30 mg/kg was intraperitoneally injected into the mice of the AS 15 and AS30 groups,respectively,and 200 mg/kg Met was given to those of the Met group by gavage,once per day,for 6 weeks.ELISA was used to detect serum testosterone(T),fasting insulin(FINS),luteinizing hormone(LH)and follicle-stimulating hormone(FSH),and the LH/FSH ratio was calculated.The levels of fasting blood glucose(FBG),triglyceride(TG)and total cholesterol(TC)were detected by automatic biochemical analyzer,and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.The estrous cycle was observed,and HE staining was performed for pathological changes in the ovary and uterus.Immunofluorescence assay was employed to measure the expression of p-eIF2α,ATF4 and CHOP in the ovarian tissue.After steroidogenic human granulosa-like tumor cell line KGN were exposed to 100 μmol/L DHEA to simulate the hyperandrogen environment of PCOS,and then treated with 5 and 10 μg/mL AS for 24 h,the protein levels of endoplasmic reticulum stress signaling pathway was detected by Western blotting.Results Compared with the Control group,the PCOS mice had disturbed estrous cycle,polycystic changes in the ovaries,and significantly increased serum T level and LH/FSH ratio(P＜0.05),and obviously elevated HOMA-IR,TC and TG levels in terms of metabolism(P＜0.01).The expression levels of p-eIF2α,ATF4 and CHOP were notably up-regulated in the ovarian granulosa cells of PCOS mice and KGN cells after DHEA exposure(P＜0.05).Additionally,AS treatment attenuated the pathological changes of ovary and uterine expression,decreased the serum T level and the LH/FSH ratio(P＜0.05),and reduced HOMA-IR,TC and TG levels(P＜0.05)when compared with the PCOS mice.Moreover,the expression levels of p-eIF2α,ATF4 and CHOP were significantly down-regulated after AS treatment in both ovarian granulosa cells of PCOS mice and KGN cells(P＜0.05).Conclusion AS significantly improves glycolipid metabolic disorder and reproductive dysfunction in PCOS mice,which may be associated with its suppressing endoplasmic reticulum stress by inhibiting the PERK/eIF2α/ATF4/CHOP pathway.

Objective To investigate the role of p300 in lipid metabolism disorders.Methods Bioinformatics analysis was performed to analyze the expression patterns of p300 in lipid metabolism disorder-related diseases and its correlation with SREBP-1c and downstream lipid metabolic enzymes.Immunofluorescence assay was used to detect the expression of p300 in the liver tissues of the patients with varying disease severity of non-alcoholic fatty liver disease(NAFLD).A mouse model of lipid metabolism disorder was established in male C57BL/6J mice by feeding high-fat diet(HFD)for 12 weeks.Western blotting was employed to assess p300 expression level in the liver tissues of HFD-fed mice.A cell model of lipid metabolism disorder was established in HepG2/AML-12 cells induced with free fatty acid(FFA).The effects of siRNA-mediated knockdown of p300 was observed to measure the levels of intracellular total cholesterol(TC)and triglyceride(TG),lipid deposition,and production of reactive oxygen species(ROS).Results Clinically,p300 was highly expressed in lipid metabolism disorders,and its level was positively correlated with NAFLD severity(P<0.05).Gene Set Enrichment Analysis(GSEA)revealed that p300 expression was significantly associated with fatty acid metabolism,cholesterol homeostasis,lipogenesis,PPAR signaling pathway,and peroxisome pathway.In vivo,p300 was significantly up-regulated in the livers of HFD-fed mice(P<0.01).In vitro,FFA stimulation markedly increased p300 expression in both HepG2 and AML-12 cells(P<0.01),whereas p300 knockdown significantly reduced intracellular TG and TC levels(P<0.01),attenuated lipid droplet accumulation,and reversed FFA-induced ROS elevation(P<0.01).Furthermore,p300 expression was positively correlated with the expression of SREBP-1c and its downstream key lipid synthesis enzymes.Conclusion p300 may promote hepatic lipid accumulation by acetylating and activating SREBP-1c and regulating downstream lipid metabolic enzymes,thereby affecting lipid synthesis and oxidative stress.These findings suggest that p300 may be a potential therapeutic target for lipid metabolism disorder-related diseases.

Objective To apply PDCA(plan,do,check and action)cycle to optimize management and shorten the turnaround time(TAT)of five sex hormone tests for improving clinical efficiency and patient satisfaction.Methods A PDCA management team was established to retrospectively analyze the current status of sex hormone TAT in 2021.In 2022,PDCA tools were utilized,including fishbone diagrams for cause analysis,Pareto charts for root cause identification,formulation and implementation of corresponding im-provement measures and monitoring of TAT changes before and after PDCA.Results Through the corrective measures,such as staff personnel training,increasing testing equipment,optimization process and other improvement measures,the median TAT of sex hor-mone samples in 2022(post-PDCA improvement)was 54 minutes which was significantly lower than 61 minutes in 2021(pre-PDCA)(P＜0.05).Conclusion PDCA cycle management effectively shortened the TAT of sex hormone samples,enhanced clinical diagnostic efficiency and patient experience,improved doctor-patient relationships,while also elevated the quality management standards of labo-ratories.

ObjectiveTo perform a predictive analysis of the quality marker（Q-Marker） for the anticoagulant activity of Kunning granules. MethodThe chemical components of Kunning granules were analyzed by ultra high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry（UHPLC-Q-TOF-MS/MS） on a Waters ACQUITY UPLC HSS T3 column（2.1 mm×100 mm， 1.8 μm） with the mobile phase of acetonitrile（A）-25 mmol∙L^-1 ammonium acetate aqueous solution（B） for gradient elution （0-5 min， 5%-22%A； 5-10 min， 22%-30%A； 10-15 min， 30%-95%A； 15-20 min， 95%-5%A； 20-30 min， 5%A）， flow rate of 0.2 mL∙min^-1， column temperature at 30 ℃， injection volume of 1 μL， electrospray ionization（ESI）， positive and negative ion detection modes. Interaction analysis between the targets of chemical components and the targets of abnormal uterine bleeding（AUB） was performed by network pharmacology， and the key components were screened through network topology analysis. The fingerprints of 10 batches of Kunning granules were established by high performance liquid chromatography（HPLC）， the anticoagulant activity of the granules was determined by blood coagulation method and fibrinogen plate method， and the spectrum-effective relationship was established. The components co-occurring in the topological analysis and spectrum-effective relationship were selected as Q-Markers， and their anticoagulant activities were verified and confirmed. ResultA total of 475 chemical components were identified from Kunning Granule， of which 22 key components such as salvianolic acid B， paeoniflorin， naringin and neohesperidin， were the potential material basis for the treatment of AUB. The spectrum-effective analysis showed that peaks 7（paeoniflorin）， 9（naringin）， 10（neohesperidin） and 11（salvianolic acid B） were the optimal principal components， and in vitro activity test showed that these four components could better characterize their anticoagulant activity. ConclusionSalvianolic acid B， paeoniflorin， neohesperidin and naringin may be Q-Markers for the anticoagulant activity of Kunning granules.

Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.
Male ; Mice ; Animals ; Cellular Reprogramming/genetics* ; Tetraploidy ; Pluripotent Stem Cells/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; DNA Methylation ; Spermatogonia/metabolism* ; Germ Cells/metabolism*

Objective To investigate the clinicopathological characteristics and independent risk factors of hepatocellular carcinoma (HCC) differentiation.  Methods A total of 108 HCC patients who underwent operation and treatment were reviewed and classified into low differentiation group (n= 29, 26.85%), medium differentiation group (n=53, 49.07%) and high differentiation group (n=26, 24.07%) according to pathological diagnosis. The clinicopathological characteristics and the expression levels of Ki67 and P53 in each group were compared and analyzed. Logistic regression model was used to analyze the risk factors for low differentiation of HCC.  Results The proportion of cirrhosis, the positive rate of P53 and Ki67 expression level in different degrees of HCC differentiation were statistically significant (P<0.05). However, there were no significant differences in age, sex, tumor diameter, and hepatitis virus infection or tumor involvement of liver capsule between groups (P>0.05). Multivariate logistic analysis showed that cirrhosis (OR=3.408), high expression of Ki67 (OR=11.113) and positive P53 (OR=9.711) were the main risk factors for poorly differentiated HCC.  Conclusion There are differences in clinical characteristics and expressions of Ki67 and P53 in HCC patients with different degrees of differentiation. Logistic regression analysis can identify clinicopathological risk factors affecting the degree of differentiation of HCC, which can provide criterion support for accurate diagnosis and prognostic treatment.

INTRODUCTION: Childhood radiation exposure is a known risk factor for thyroid malignancy and dysfunction. However, local data are limited and there is no consensus on the modality and frequency of screening in this high-risk group. METHODS: Retrospective analysis study evaluating patients with childhood radiation exposure in 2006-2016 and minimum of 1-year follow-up. RESULTS: Of the 132 childhood cancer survivors in the study, thyroid malignancy was detected in 2 cases (1.5%) and thyroid nodules in 13 (9.8%). The earliest thyroid malignancy was detected 5 years post-radiotherapy via ultrasound. Of the 84 patients who had screening thyroid function test, 26 (31.0%) were detected with abnormal test results post-radiation, majority being subclinical hypothyroidism. CONCLUSION Regular screening via clinical examination for thyroid nodules should be performed at least annually. Where feasible and if resources permit, consideration should be given to using ultrasound for thyroid nodule(s) and malignancy screening at 5 years post-radiation therapy. Screening for thyroid dysfunction can be considered from 6-12 months post-radiotherapy.

Introduction: Derangement in calcium homeostasis is common in nephrotic syndrome (NS). It is postulated that low serum total calcium and vitamin D levels are due to loss of protein-bound calcium and vitamin D. It is unclear if free calcium and free vitamin D levels are truly low. The guideline is lacking with regards to calcium and vitamin D supplementation in NS. This study aims to examine calcium and vitamin D homeostasis and bone turnover in NS to guide practice in calcium and vitamin D levels supplementation. Methodology: This is a prospective pilot study of ten patients diagnosed with NS, and eight healthy controls. Calcium, vitamin D, and bone turnover-related analytes were assessed at baseline, partial and complete remission in NS patients and in healthy controls. Results: NS patients had low free and total serum calcium, low total 25(OH)D, normal total 1,25(OH)D levels and lack of parathyroid hormone response. With remission of disease, serum calcium and vitamin D metabolites improved. However, nephrotic patients who do not attain complete disease remission continue to have low 25(OH)D level. Conclusion In this study, the vitamin D and calcium derangement observed at nephrotic syndrome presentation trended towards normalisation in remission. This suggested calcium and vitamin D replacement may not be indicated in early-phase nephrotic syndrome but may be considered in prolonged nephrotic syndrome.
Vitamin D Deficiency

Brexanolone, also known as 3a, 5a-tetrahydroprogesterone, is a metabolite of progesterone and an important neuroactive steroid with a wide range of biological activities.Recently, the most exciting thing is that 2019 year the FDA has approved its treatment of postpartum depression, which is also the only drug to obtain indications for postpartum depression.It is expected to officially enter the market by the end of June 2019 year.This article reviews the research progress of its pharmacological action.

Objective To explore the value of cMRI and MRS in diagnosis and differential diagnosis of single brain metastases and localized high grade gliomas.Methods The cMRI (T1WI,T2WI and contrast-enhanced MRI)and MRS data of 23 cases with single brain metastases and in 28 cases with high grade gliomas confirmed by pathology were collected and analyzed retrospectively.Results cMRI:①There were no significant differences of T1WI,T2WI and constrast-enhanced MRI between single brain metastases and high-grade gliomas.②Enhanced images of single brain metastases and localized high-grade gliomas showed that there was a significant difference in the peritumoral edema area:the nodular or ring enhancement were found in single brain metastases,the outer outline of ring enhancement was smooth,and there was no enhancement at the peripheral edema area;Irregular patchy or ring enhancement were found in localized high-grade glioma tumor,both of the outer and inner edges of the ring enhancement rough,a little patchy enhancement at the peripheral edema area were found in 1 6 patients,and there was no enhancement of the edema area in the other 1 2 patients.MRS:①There were no significant differences of the tumor parenchyma in the peaks of NAA,Cr,Cho and central Lac between single brain metastases and localized high grade gliomas.②A significant difference of the peripheral edema area were observed between them.The peaks of NAA,Cr,Cho at the edema area in all single brain metastases patients were normal.But in all high-grade glioma patients,the NAA and Cr peaks were decreased,while the Cho peaks were increased.Conclusion The morphological manifestations of single brain metastases are similar to localized high grade gliomas.An obvious difference of cMRI and MRS lines exists in the peritumoral edema area between them,which could be used for differential diagnosis.