BACKGROUND:Studies have shown that patients with premature ovarian failure have changes in the structure of intestinal flora and that imbalance of intestinal microbiota may be one of the important mechanisms in the development of premature ovarian failure. OBJECTIVE:To investigate the effect of Liuwei Dihuang Wan on oxidative stress and intestinal microbiota in premature ovarian failure mice induced by cyclophosphamide. METHODS:Forty-five female ICR mice were randomized into three groups:blank group(normal mice),model group(premature ovarian failure mice),and Liuwei Dihuang Wan group.A mouse model of premature ovarian failure was prepared by one-time intraperitoneal injection of cyclophosphamide(120 mg/kg)in the latter two groups.After successful modeling,the Liuwei Dihuang Wan group was intragastrically administered for 28 continuous days,and the other two groups were intragastrically administered with the same amount of normal saline for 28 days.Mouse body mass was recorded weekly and ovarian index was calculated.The development of mouse follicles was observed using hematoxylin-eosin staining.ELISA method was used to detect serum levels of anti-Mullerian hormone,estradiol,follicle stimulating hormone,superoxide dismutase,glutathione peroxidase,and malondialdehyde.Meanwhile,the gut microbiome of all mice was detected through 16S rDNA sequencing. RESULTS AND CONCLUSION:The mice in the model group had loose hair,decreased vigor and grip strength,almost no increase in body mass,and decreased ovarian index.Whereas,the mouse body mass and ovarian index were increased after treatment with Liuwei Dihuang Wan(P<0.05).The estrous cycle of mice in the model group was disorganized;Liuwei Dihuang Wan could restore the estrous cycle and reduce the number of atretic follicles in mice with premature ovarian failure.The serum levels of follicle stimulating hormone and malondialdehyde in the model group significantly increased(P<0.01),while the levels of estradiol,anti-Mullerian hormone,superoxide dismutase,and glutathione peroxidase significantly decreased(P<0.01).Liuwei Dihuang Wan could significantly decrease the serum levels of follicle stimulating hormone and malondialdehyde(P<0.01),and increase the levels of estradiol,anti-Mullerian hormone,superoxide dismutase,and glutathione peroxidase.According to the 16S rDNA sequencing results,Liuwei Dihuang Wan could regulate the abundance and diversity of intestinal microbiota,and increase the relative abundance of beneficial bacteria.KEGG pathway analysis showed that the intestinal microbiota and metabolic pathways,biosynthesis of secondary metabolites,microbial metabolism in different environments,and biosynthesis of amino acids were regulated by Liuwei Dihuang Wan.To conclude,the changes in the structure of intestinal microbiome may be one of the potential mechanisms of Liuwei Dihuang Wan in treating premature ovarian failure.Liuwei Dihuang Wan can regulate the structure of intestinal microbiome,increase the number of beneficial bacteria,reduce the number of harmful bacteria,and thus improve the balance of intestinal microbiota.This regulatory effect helps to reduce oxidative stress levels and further inhibit ovarian oxidative stress in mice with premature ovarian failure.

BACKGROUND:The granulosa cell state greatly affects the quality of oocytes.As the increased proportion of aged women in assisted reproduction,granulosa cells from patients of different ages are examined at transcript levels in order to better assess oocyte quality. OBJECTIVE:To investigate the expression changes of mRNA and long non-coding RNA in granulosa cells from the young and aged patients. METHODS:Waste cumulus granulosa cells obtained during the assisted reproductive cycle.Young group was in 25-35 years old,and the aged group was in 38-45 years old.Granulosa cells collected from the young and aged females were analyzed using RNA sequencing,three replications for each group. RESULTS AND CONCLUSION:(1)The RNA sequencing analysis results showed that the average sequencing volume of samples was more than 14 G;the data quality Q30 after quality control was more than 93%;the average mapping rate of data was 98.4%,which indicates that the data had good quality.(2)The results of principal component analysis and correlation analysis showed that the samples of the aged group and the young group could be clearly distinguished.(3)The difference analysis results showed that compared with the young group,a total of 410 differentially expressed mRNAs were detected in the aged group(167 up-regulated and 243 down-regulated).GO analysis results showed that the down-regulated genes were mainly enriched in regulatedexocytosis and the emetabolicprocess.GSEA analysis results showed that secretion-related pathways in the aged group were down-regulated.(4)Compared with the young group,662 differentially expressed long non-coding RNAs were detected in the aged group,and 1 772 protein-coding genes were directly regulated by these long non-coding RNAs,among which 59 genes overlaped with differentially expressed genes.The results showed that the expression of secretion-related genes and pathways in granulosa cells of the aged group was down-regulated,thus affecting oocyte quality.At the same time,these down-regulated genes were regulated by long non-coding RNA.Therefore,the expression of long non-coding RNA might be related to age.

ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy， to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian， and meanwhile， the contents of four index components（geniposide， paeoniflorin， hesperidin and paeonol） were quantitatively analyzed by high performance liquid chromatography（HPLC）. Nonalcoholic fatty liver disease（NAFLD） rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group， model group， lovastatin group（1.8 mg·kg^-1）， combined decoction group（1.26 g·kg^-1） and single decoction group（1.18 g·kg^-1）. After successful modeling， lovastatin group， combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day， and the control group and model group were given equal amounts of normal saline by intragastric administration， after 4 weeks of administration， the serum and liver tissues were collected， and the contents of alanine aminotransferase（ALT）， aspartate aminotransferase（AST）， total cholesterol（TC）， triglyceride（TG）， low-density lipoprotein cholesterol（LDL-C） and high-density lipoprotein cholesterol（HDL-C） in serum of rats were detected， and the liver pathological examination was carried out by hematoxylin-eosin（HE） staining and oil red O staining， so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian， and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide， paeoniflorin， hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples（P<0.01）. The pharmacodynamic results showed that compared with the model group， both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats， reduce the serum levels of AST， ALT， TC， TG and LDL-C， increase the serum level of HDL-C， and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However， there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition， but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.

Objective: To evaluate the clinical efficacy of digitally guided precision crown lengthening in secondary aesthetic rehabilitation cases, and to provide a clinical reference for digitally guided crown lengthening procedures and secondary aesthetic restorations. Methods: We present a case of a patient with tetracycline-stained teeth, partial detachment of anterior resin veneers, and gingival margin discrepancies. The patient underwent digitally guided precision crown lengthening followed by secondary aesthetic rehabilitation. Multimodal data, including intraoral, facial, and CBCT scans, were integrated to construct a four-dimensional virtual patient model (incorporating teeth, face, bone, and occlusion) for surgical planning and 3D-printed guide fabrication. Secondary aesthetic restoration was performed after achieving stable post-surgical outcomes. Based on this case, we conducted a detailed analysis and reviewed relevant literature on crown lengthening in secondary aesthetic rehabilitation. Results: The gingival contour of the anterior teeth exhibited significant improvement, with enhanced symmetry and stable gingival margin positioning that closely matched the preoperative design. The crown lengthening procedure demonstrated high precision, and the final outcome was aesthetic and functional. Literature review indicated that secondary restorations frequently present challenges such as gingival contour discrepancies and inflammation. Aesthetic crown lengthening in the anterior region should optimize both soft and hard tissue morphology to meet aesthetic standards, with digital technology improving procedural accuracy. Conclusion Precision crown lengthening effectively addresses gingival margin discrepancies in secondary aesthetic rehabilitation, ensuring stable gingival positioning and superior aesthetic outcomes. This approach is particularly suitable for cases with high aesthetic demands.

Aim To investigate the effect of ellagic acid (EA) on cognitive function in APP/PS 1 double- transgenic mice, and to explore the regulatory mechanism of ellagic acid on the level of oxidative stress in the hippocampus of double-transgenic mice based on the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3 (PI3K/AKT/GSK-3 β) signaling pathway. Methods Thirty-two SPF-grade 6-month-old APP/PS 1 double transgenic mice were randomly divided into four groups, namely, APP/PS 1 group, APP/PS1 + EA group, APP/PS1 + LY294002 group, APP/PS 1 + EA + LY294002 group, with eight mice in each group, and eight SPF-grade C57BL/6J wild type mice ( Wild type) were selected as the blank control group. The APP/PS 1 + EA group was given 50 mg · kg

ObjectiveTo describe the epidemic characteristics of COVID-19 after policy adjustment from “Category B notifiable disease with category A management” to “Category B notifiable disease with category B management”， and to explore the protective effect of previous infection with SARS-CoV-2 on common symptoms of reinfection. MethodsHealthcare workers infected with SARS-CoV-2 in a grade A tertiary hospital in Shanghai were included in the study from December 4， 2022 to January 11， 2023. Data on demographic characteristics， clinical symptoms， medical history， and COVID-19 vaccination history were collected. We determined the epidemiological curve and characteristics， and then compared the difference in the severity of clinical symptoms between primary and reinfection subjects. ResultsA total of 2 704 cases were included in the study， of which 45 had reinfection， 605 （22.4%）were males， 608 （22.5%）were doctors， 1 275 （47.2%） were nurses， and 2 351 （86.9%） received ≥3 doses of COVID-19 vaccination. The average age of these healthcare workers was （34.9±9.1） years old. The number of cases with mild/moderate illness， asymptomatic infection， fever， headache， dry cough， expectoration， and chest tightness were 2 704 （100.0%）， 92 （3.4%）， 2 385 （88.2%）， 2 066 （76.4%）， 1 642 （60.7%）， 1 807 （66.8%）， and 439 （16.2%）， respectively. Reinfection was a protective factor for fever （OR=0.161， P<0.001）， headache （OR=0.320， P<0.001）， and peak body temperature （β=-0.446， P<0.001）. ConclusionFollowing the COVID-19 policy adjustment as a category B notifiable disease， healthcare workers at a grade A tertiary hospital in Shanghai predominantly experiences mild to moderate COVID-19 symptoms. Reinfection results in milder clinical manifestations， with a lower proportion of being asymptomatic.

Objective To clarify the effects of 5G mobile phone radiofrequency radiation exposure on male mouse fertility and to preliminarily explore the underlying mechanisms. Methods Healthy male C57BL/6 mice aged 7-8 weeks were randomly assigned to Sham group, 3.5 GHz radiofrequency radiation group, and 4.9 GHz radiofrequency radiation group, with 16 mice in each group. The mice were exposed to 3.5 GHz or 4.9 GHz mobile phone radiofrequency radiation for 42 consecutive days (1 h per day). The sperm quality was evaluated using sperm count, deformity rate, and motility. H&E staining was performed to assess testicular tissue structure by observing the morphology of spermatogenic cells at various development stages, the diameter of seminiferous tubules, and the thickness of seminiferous epithelium. The sperm mitochondrial function was assessed using sperm mitochondrial membrane potential and testicular ATP content. The fertility of mice was evaluated through fertility rate, litter size, and survival rate of offspring. The underlying mechanisms were explored by detecting the methylation of LRGUK gene and its mRNA and protein levels. Results Compared with the Sham group, there were no significant changes in sperm count in the 3.5 GHz and 4.9 GHz groups; however, the sperm abnormality rate significantly increased (P < 0.05) and sperm motility significantly decreased (P < 0.05). The structure of testicular tissue, the function of sperm mitochondria, and fertility of mice showed no significant changes as compared with the Sham group. The methylation level of LRGUK gene in the testes significantly increased, while the mRNA and protein expression levels significantly decreased. Conclusion Exposure to 3.5 GHz and 4.9 GHz mobile phone radiofrequency radiation for 42 consecutive days can lead to an increase in sperm deformity rate and a decrease in sperm motility in mice, but has no significant effect on fertility, which may be related to an increase in methylation level of the LRGUK gene in the testes.

Objective To investigate the effect of transmembrane protein CD147 expression on AIM2 inflammasome-mediated pyroptosis and proliferation of cervical cancer cells.Methods Western Blot was used to detect the expression of CD147 in cervical cancer cell lines SiHa(HPV+)and C33a(HPV-)and normal cervical epithelial cells H8(HPV+)and HCer Epic(HPV-).SiHa cells were transfected with lentivirus to down-regulate the expression of CD147.According to the different treatments,SiHa cells were divided into SiHa group,negative control group(shCD147-NON),knockdown group 1(shCD147-1)and knockdown group 2(shCD147-2).The transfection effect was verified by Western Blot,RT-qPCR and green fluorescence expression.The protein and mRNA expressions of AIM2,Caspase-1,IL-18 and GSDMD were detected by Western Blot and RT-qPCR.The lactate dehydrogenase(LDH)release was measured in the cell culture supernatant,and the cell morphology was observed under the fluorescence inverted microscope;the proliferation ability of cells was measured by CCK-8 and the colony formation ability was measured by cell cloning experiments.Results Western Blot results showed that CD147 protein expression in SiHa cells was the highest compared with that in HCerEpic cells.CD147 low expression lentivirus effectively down-regulated the expression of CD147 in SiHa cells.The results of Western Blot and RT-qPCR experiments showed that the expression of AIM 2,Caspase-1,IL-18,GSDMD protein and mRNA increased in shCD147-1 and shCD147-2 group(P<0.05).Lactate dehydrogenase(LDH)release assay showed that compared with the SiHa group,the shCD147 group had a significant increase in LDH release(P<0.05).Fluorescence inverted microscope showed that the shCD147 group had swelling and vacuolization,showing typical pyroptosis.Compared with the SiHa group,the shCD147-1 and shCD147-2 groups had significantly reduced the cell proliferation and colony formation ability(P<0.05).Conclusion Low expression of CD147 effectively up-regulates the expression of AIM2 inflammation-related factors in cervical cancer SiHa cells,induces the pyroptosis,and inhibits the cell proliferation and cloning.

Objective: To construct a prediction model for preeclampsia (PE) risk in twin-pregnant women, so as to provide the basis for early screening and prevention of PE. Methods: A total of 467 twin-pregnant women who underwent prenatal examination and delivered at Huzhou Maternal and Child Health Hospital were selected. Sixty cases with preeclampsia (PE) were included in the case group, and 60 women without PE were included in the control group. General information, blood biochemical indicators and uterine artery resistance index (UtA-RI) were collected. A logistic regression model was used to screen predictive factors and establish a nomogram. The Bootstrap method was performed for the internal validation; the receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis were employed to evaluate the discrimination, calibration and clinical utility of the nomogram, respectively. Results: In the case group, there were 47 individuals (78.33%) aged younger than 35 years, 21 individuals (35.00%) with pre-pregnancy body mass index (BMI) of 25 kg/m2 and above, and 33 individuals (55.00%) with in vitro fertilization. In the control group, there were 57 individuals (95.00%) aged younger than 35 years, 8 individuals (13.33%) with pre-pregnancy BMI of 25 kg/m2 and above, and 39 individuals (65.00%) with natural pregnancy. Multivariable logistic regression analysis identified age, pre-pregnancy BMI, method of conception, placental growth factor (PLGF) and UtA-RI as risk prediction factors for PE in twin-pregnant women. The established nomogram had an area under the ROC curve of 0.827 (95%CI: 0.755-0.899), a sensitivity of 0.767, a specificity of 0.733, a good discrimination and calibration, and a relatively high clinical net benefit. Conclusion The nomogram established by age, pre-pregnancy BMI, method of conception, PLGF and UtA-RI has a good predictive value for the risk of PE in twin-pregnant women.

Objective To evaluate the clinical efficacy and safety of vitamin D combined with atomoxetine hydrochloride in the treatment of attention deficit hyperactivity disorder(ADHD)in children.Methods A retrospective analysis was performed for the data of children with ADHD admitted to Huzhou First People's Hospital from September 2022 to September 2023,and the children were divided into combination group and control group according to the diagnosis and treatment plan.The efficacy of treatment and cognitive function[Mini-Mental State Examination Scale(MMSE)and Wechsler Intelligence Scale for Children,Fourth Edition(WISC-IV.)],behavioral dysfunction[Conners]and neurofilament markers[neurential-specific enolase(NSE)and neurofilament light chain protein(NFL))before and after 3 months of treatment were observed and compared after treatment.The occurrence of adverse reactions during the treatment period was observed and recorded,and the safety was evaluated.Results A total of 101 children with ADHD were enrolled,including 51 in the combined group and 50 in the control group.After treatment,the total effective rate of the combination group was significantly higher than that of the control group(P<0.05).Before treatment,there were no significant differences in MMSE score,WISC-IV.score,Conners score,serum NSE level and serum NFL level between the two groups(P>0.05).After treatment,the MMSE score,WISC-IV.score,Conners score,serum NSE level and serum NFL level of the two groups were significantly improved compared with those before treatment(P<0.05),and the improvement degree in the combination group was better than that in the control group(P<0.05).There was no significant difference in the incidence of adverse reactions between the two groups during treatment(P>0.05).Conclusion Compared with atomoxetine hydrochloride monotherapy,vitamin D combined with atomoxetine hydrochloride has better efficacy in children with ADHD,and can improve cognitive function,behavioral dysfunction and neurological impairment,with high safety.