Non-alcoholic fatty liver disease (NAFLD) is a metabolic disease characterized by abnormal deposition of lipid in hepatocytes. If not intervened in time, NAFLD may develop into liver fibrosis or liver cancer, and ultimately threatening life. NAFLD has complicated etiology and pathogenesis, and there are no effective therapeutic means and specific drugs. Currently, insulin sensitizers, lipid-lowering agents and hepatoprotective agents are often used for clinical intervention, but these drugs have obvious side effects, and their effectiveness and safety need to be further confirmed. Adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays a central role in maintaining energy homeostasis. Activated AMPK can enhance lipid degradation, alleviate insulin resistance (IR), suppress oxidative stress and inflammatory response, and regulate autophagy, thereby alleviating NAFLD. Natural herbal medicines have received extensive attention recently because of their regulatory effects on AMPK and low side effects. In this article, we reviewed the biologically active natural herbal medicines (such as natural herbal medicine formulas, extracts, polysaccharides, and monomers) that reported in recent years to treat NAFLD via regulating AMPK, which can serve as a foundation for subsequent development of candidate drugs for NAFLD.

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 explore the role of prognostic nutritional index (PNI) and pleural thickness in the prognostic evaluation of patients with epithelial malignant pleural mesothelioma (MPM) .Methods:In April 2022, a retrospective analysis was conducted on the data and laboratory data of 41 patients with epithelial MPM admitted to the cardiothoracic surgery department of Chuxiong Yi Autonomous Prefecture People's Hospital from January 2018 to May 2021. Univariate and multivariate analysis were used to evaluate the relationships between total survival time, clinical characteristics, PNI and pleural thickness in patients.Results:The 41 patients were mostly male (26 cases, 63.4%) , with a median age of 55 years old. The main clinical manifestations were chest pain (53.7%) , bloody pleural effusion (75.6%) , and chest pain combined with bloody pleural effusion (36.6%) . The median survival time of patients with different TNM stage, efficacy after 4 cycles of chemotherapy, PNI, maximum pleural thickness after chemotherapy (post max) , sum of post max in 3 zones after chemotherapy (post sum) were statistically different (χ 2=3.89, 14.51, 15.33, 4.33, 12.05, P<0.05) . Compared with patients with high PNI and post sum<32.26 mm, MPM patients with low PNI and post sum≥32.26 mm have higher risk of death, and the differences were statistically significant ( HR=1.52, 95% CI: 1.75-11.93, P=0.002; HR=1.70, 95% CI: 1.84-16.23, P=0.002) . Conclusion:PNI and post sum can be used to predict the prognosis of patients with epithelial MPM.

Objective:To explore the role of prognostic nutritional index (PNI) and pleural thickness in the prognostic evaluation of patients with epithelial malignant pleural mesothelioma (MPM) .Methods:In April 2022, a retrospective analysis was conducted on the data and laboratory data of 41 patients with epithelial MPM admitted to the cardiothoracic surgery department of Chuxiong Yi Autonomous Prefecture People's Hospital from January 2018 to May 2021. Univariate and multivariate analysis were used to evaluate the relationships between total survival time, clinical characteristics, PNI and pleural thickness in patients.Results:The 41 patients were mostly male (26 cases, 63.4%) , with a median age of 55 years old. The main clinical manifestations were chest pain (53.7%) , bloody pleural effusion (75.6%) , and chest pain combined with bloody pleural effusion (36.6%) . The median survival time of patients with different TNM stage, efficacy after 4 cycles of chemotherapy, PNI, maximum pleural thickness after chemotherapy (post max) , sum of post max in 3 zones after chemotherapy (post sum) were statistically different (χ 2=3.89, 14.51, 15.33, 4.33, 12.05, P<0.05) . Compared with patients with high PNI and post sum<32.26 mm, MPM patients with low PNI and post sum≥32.26 mm have higher risk of death, and the differences were statistically significant ( HR=1.52, 95% CI: 1.75-11.93, P=0.002; HR=1.70, 95% CI: 1.84-16.23, P=0.002) . Conclusion:PNI and post sum can be used to predict the prognosis of patients with epithelial MPM.

Objective:To observe and compare the clinical characteristics between post-chronic pancreatitis diabetes mellitus(PPDM-C)patients and type 2 diabetes mellitus(T2DM).Methods:Data of 142 cases of CP patients confirmed in Shanghai Pudong New Area Gongli Hospital from January 2018 to December 2021 were collected, all the patients were divided into CP group without diabetes mellitus ( n=60) and PPDM-C group with diabetes mellitus ( n=82) based on whether with or without diabetes mellitus. And 82 cases T2DM without CP (T2DM group, n=82) hospitalized simultaneously were collected as control group. The age, sex, body mass index, onset characteristics, laboratory examination indicators at admission (fasting blood glucose, glycosylated hemoglobin, blood creatinine, and alanine transaminase), imaging characteristics of the pancreas (pancreatic atrophy, multiple calcifications of the pancreas, pancreatic duct stones, pancreatic duct dilation, and pancreatic duct obstruction), and treatments and efficacy of diabetes were recorded. Results:Compared with T2DM group, PPDM-C group had lower body mass index (22.2 kg/m 2vs 24.6 kg/m 2), and glycosylated hemoglobin levels (7.34% vs 9.20%) (all P values <0.001), higher alanine transaminase levels (33.00 U/L vs 18.65 U/L, P =0.021). And they had more upper abdominal pain, nausea, vomiting, weight loss and diarrhea symptoms. In addition, they had less use of combination of insulin and hypoglycemic drugs to control blood glucose. And compared with CP group, PPDM-C group had higher body mass index (22.06 kg/m 2vs 21.18 kg/m 2), higher glycosylated hemoglobin levels (7.34% vs 5.70%), higher fasting blood-glucose levels (7.91 mmol/l vs 5.31 mmol/l), higher alanine transaminase levels (33.00 U/L vs 26.50U/L), and their differences were statistically significant (all P values <0.05). And they had higher incidence of pancreatic atrophy, multiple calcifications in the pancreatic duct and pancreatic duct obstruction (all P values <0.05). Conclusions:PPDM-C patients are more likely to experience digestive system symptoms such as abdominal pain than T2DM patients, while their pancreatic malfunction is more likely to occur compared to CP patients. More attentions to PPDM-C associated clinical manifestations, biochemical and imaging changes could identify patients at potential risk for early diagnosis and treatment earlier.

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*

ObjectiveTo investigate the clinical and prognostic features of hypertriglyceridemic pancreatitis (HTGP) during pregnancy. MethodsA total of 77 patients with acute pancreatitis in pregnancy who were admitted to The Affiliated Hospital of Southwest Medical University from January 2013 to June 2020 were enrolled as subjects, and according to the presence or absence of HTGP, they were divided into HTGP group with 38 patients and non-HTGP group with 39 patients. The two groups were compared in terms of the clinical features including age, pregnancy, laboratory markers, complication, and prognostic indices. The independent samples t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups, and the Mann-Whitney U test was used for comparison of ranked data between two groups. According to the trimester of pregnancy, the patients were divided into early pregnancy group with 4 patients, middle pregnancy group with 14 patients, and late pregnancy with 59 patients, and blood lipid levels were compared between groups; the Kruskal-Wallis H test was used for comparison between two groups or within each group. ResultsCompared with the non-HTGP group, the HTGP group had significantly higher mean age (t=-3.000, P=0.004) and proportion of patients with diabetes (χ2=8.867, P=0.003). Compared with the non-HTGP group, the HTGP group had a significantly higher proportion of patients with disease onset in the third trimester (P＜0.001) and a significantly lower serum level of sodium (Z=-6.328, P＜0.001). Compared with the non-HTGP group, the HTGP group had significantly higher incidence rates of acute peripancreatic fluid collection (76.3% vs 33.3%, χ2=14.342, P＜0.001) and systemic inflammatory response syndrome (81.6% vs 410%, χ2=13.310, P＜0.001). There were significant differences between the two groups in the length of hospital stay, the rate of admission to the intensive care unit, and hospital costs (Z=-2.077, χ2=11.537, Z=-3.525, all P＜0.05). The levels of total cholesterol and triglyceride in the third trimester were significantly higher than those in middle pregnancy (both P＜0.05). ConclusionCompared with the non-HTGP group, the HTGP group has a higher proportion of patients with disease onset in the third trimester of pregnancy, a higher risk of complications, a longer length of hospital stay, and higher hospital costs.

BACKGROUND: Increasing attention has been paid to vascular components of the adipose-derived matrix and adipose-derived stem cells in tissue engineering. Existing methods for separating the vascular components of the adipose-derived matrix mainly include enzymatic and bolus injection, both of which have fatal disadvantages. OBJECTIVE: To search for a method for preparing adipose-derived stromal vascular fractions with high efficiency, safety, and simplicity. METHODS: The group without any treatment was used as the negative control, and the enzymatic hydrolysis method served as the positive control. The enzymatic hydrolysis method, traditional bolus method, modified bolus method, glass beads method and built-in ultrasonic waves method were compared through cell volume, survival rate, cell fragments, cell viability, increment rate and detection of microbial infection. The enzymatic hydrolysis method and the common bolus injection method were commonly used in the separation of vascular component cells of the fat source matrix; the improved bolus method was a method obtained by improving on the basis of the ordinary bolus method; the glass bead method was to use the glass bead to oscillate. The shear force generated was obtained by adding glass beads to the fat granules and shaking at 2 500 r/min for 9 minutes to prepare stromal vascular fraction cells. Using the built-in ultrasonic method, adipose tissue was treated at 25 W for 36 seconds to obtain stromal vascular fraction cells through a cavitation effect. RESULTS AND CONCLUSION: (1) The size of stromal vascular fraction cells isolated by five methods showed no significant difference (P > 0.05). (2)The cell viability was lowest in the negative control group, and highest in the enzymatic hydrolysis group. The cell viability in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups was significantly higher than that in the modified and traditional bolus groups (P < 0.05). (3) The cell survival rate and cell proliferation rate in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups were significantly higher than those in the modified and traditional bolus groups (P < 0.05). (4) The cell fragmentation rate and cell apoptosis rate in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups were significantly lower than those in the modified and traditional bolus groups (P < 0.05). (5) These results indicate that the built-in ultrasonic method and the glass bead method are better in enriching vascular components of the adipose-derived matrix. But glass bead method adds exogenous products, so it increases the risk of pollution. Built-in ultrasonic method inserts the ultrasound probe into the adipose tissue, but as long as the ultrasound probe is thoroughly sterilized, the risk of contamination is minimized. In general, the built-in ultrasonic method and the glass bead method are superior to modified and traditional bolus methods.

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.