The purpose of this study was to investigate the intervention effect and mechanism of Lycium barbarum leaves on letrozole-induced polycystic ovary syndrome (PCOS) mice. The PCOS model was prepared by letrozole combined with high-fat diet. After successful modeling, 40 mice were randomly divided into PCOS group, positive drug metformin group, low-dose Lycium barbarum leaves group, and high-dose Lycium barbarum leaves group. The corresponding drugs were given by gavage for 29 days. At the end of the experiment, the eyeballs were removed for blood collection and ovarian tissue was collected. The ovarian mass, fasting blood glucose (FBG), fasting insulin (FINS), testosterone (T), anti-Mullerian hormone (AMH), luteinizing hormone (LH), follicle stimulating hormone (FSH), and estradiol (E₂) levels were measured in each group. The morphology of ovarian tissue was observed by hematoxylin-eosin staining, and the oocytes, cystic follicles and corpus luteum were counted. Cecal contents of mice were collected for analysis of intestinal flora composition and differential flora. The animal experiment process was approved by the Animal Ethics Committee of Nanjing University of Traditional Chinese Medicine. The results showed that the estrous cycle of PCOS mice was disordered. Compared with the PCOS group, the Lycium barbarum leaves group can significantly reduce the ovarian damage of mice, reduce the number of cystic dilated follicles, and normalize the estrous cycle. After the intervention of Lycium barbarum leaves, the levels of FBG, FINS, T, AMH, LH, FSH and LH/FSH were significantly decreased (P < 0.05), while the level of E₂ was significantly increased (P < 0.001). In addition, Lycium barbarum leaves can regulate the disorder of intestinal flora diversity in PCOS mice, increase the abundance of Bacteroidetes, and reduce the abundance of Firmicutes, Ileibacterium, Romboutsia and Faecalibaculum. In summary, Lycium barbarum leaves can play a therapeutic role in PCOS mice by improving insulin resistance, regulating reproductive hormone disorders and gut microbiota imbalance. It provides scientific basis and useful reference for the rational utilization and development of Lycium barbarum leaves.

This study was aimed at understanding the trends in,and scope of,severe fever with thrombocytopenia syndrome(SFTS)in Nanjing,analyzing the spatial distribution pattern,detecting high incidence cluster areas and key popula-tions,and scientifically guiding prevention and control strategies and measures.We obtained data on SFTS cases from 2010 to 2023 in Nanjing from the China Disease Control and Prevention Information System,and described the time,popu-lation,and spatial distribution characteristics.We used joinpoint regression to calculate the annual percentage change(APC)in incidence,then used FleXScan spatial clustering scan analysis to explore spatial clustering areas at the street level.A total of 507 SFTS cases were reported from 2010 to 2023 in Nanjing.The APC was 31.8％(95％CI:22.5％-41.9％,P＜0.001),and the reported incidence in 2023 was 1.42/100 000(134 cases).The seasonal indices from May to August were 2.7,2.1,3.0,and 1.3,respectively,accounting for 76.1％of the total cases.The median age was 66(IQR:55,73)years,which gradually increased from 59 years in 2010-2011 to 68 in 2022-2023(P＜0.001);94.1％of cases were in individuals 45 years or older.Farmers,homemakers/unemployed individuals,and retirees accounted for 90.1％.The epidemic area increased from 11 streets in four districts in 2010-2011 to 58 streets in 11 dis-tricts in 2022-2023.Except for 2012-2013,global spatial autocorrelation analysis showed positive Moran's I values(0.224-0.526,P＜0.001),and FlexScan scan indicated that several streets in Lishui District and Jiangning District were the most likely clusters.Four streets in Pukou District were the secondary clusters from 2018 to 2023,and three streets in Luhe District in 2022-2023 were the secondary clusters(all P＜0.05).The reported incidence of SFTS in Nanjing showed a rapid upward trend,with spread of epidemic areas.The spatial distribution pattern was clustered.Strengthened training in diagnosis and treatment technology and detection ability of medical institutions,surveillance in high-incidence areas,tracing of case flow,and health education of tick and disease prevention knowledge are recommended.

This paper explored the protective effect and potential mechanism of Shouhui Tongbian Capsules(SHTB) on cerebral ischemia-reperfusion rat models. Rats were randomly divided into sham surgery group, model group, low-dose SHTB group(0.2 g·kg~(-1)·d~(-1)), high-dose SHTB group(SHTB g·kg~(-1)·d~(-1)), and an edaravone positive drug group(5.4 mg·kg~(-1)·d~(-1)), with 12 rats in each group. Rats were given continuous intragastric administration seven days before surgery, and the suture method was used to establish the cerebral ischemia-reperfusion injury(CIRI) rat model. Zea-Longa rating scale for neurological functions was used to assess the degree of neurological function impairment in rats; hematoxylin-eosin(HE) staining was used to observe the pathological damage of rats' brain tissue; TTC staining was used to measure the area of cerebral infarction in rats; enzyme-linked immunosorbent assay(ELISA) was used to detect the serum levels of interleukin-6(IL-6), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) in each group. Western blot was used to detect the level of tight junction protein associated with the blood-brain barrier and intestinal barrier, as well as the protein expression of the TLR4/MyD88/NF-κB pathway in brain tissue. Changes in rats' brain tissue and metabolites in serum were detected by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS), so as to explore the potential mechanism of SHTB treatment for CIRI in rats. Compared with the model group, SHTB could significantly alleviate the pathological damage to the brain of CIRI rats, reduce the volume of cerebral infarction, and lower the level of inflammation in the serum; Western blot results showed that SHTB could regulate the expression of tight junction proteins related to the blood-brain barrier and intestinal barrier in CIRI rats and downregulate the expression of TLR4, NF-κB, and MyD88 proteins in brain tissue. The UPLC-MS/MS results indicated that SHTB could significantly regulate the content of potential differential metabolites such as fatty acids, and serum and brain tissue are involved in pathways such as unsaturated fatty acid biosynthesis. SHTB could repair intestinal barrier function, reduce inflammation levels in the body, and improve the damaged blood-brain barrier, exerting a protective effect on brain nerves. Its mechanism may be achieved by balancing fatty acid metabolism and regulating the expression of TLR4/MyD88/NF-κB signaling pathway proteins.
Animals ; Rats ; Drugs, Chinese Herbal/administration & dosage* ; Reperfusion Injury/genetics* ; Male ; Rats, Sprague-Dawley ; Brain Ischemia/genetics* ; Metabolomics ; Disease Models, Animal ; Myeloid Differentiation Factor 88/metabolism* ; Toll-Like Receptor 4/metabolism* ; NF-kappa B/metabolism* ; Humans ; Brain/metabolism*

AIM: To investigate the changes in retinal nerve fiber layer(RNFL)and macular retinal thickness(MRT)in children with refractive abnormalities and amblyopia, and their predictive value of outcome.METHODS: A total of 168 children with myopic refractive abnormalities and monocular amblyopia admitted to our hospital from January 2020 to October 2022 were selected as the observation group, with 118 cases of mild to moderate amblyopia and 50 cases of severe amblyopia, and 168 children with normal vision were included as the control group in a 1:1 ratio during the same period. The changes of RNFL and MRT in two groups of children were statistically counted, and the correlation between the severity of refractive abnormalities and RNFL and MRT in children with amblyopia was analyzed. Additionally, the observation group was divided into effective subgroup and ineffective subgroup based on the therapeutic effect. The general information, as well as RNFL and MRT of the effective subgroup and the ineffective subgroups before and after treatment were compared. Logistic was used to analyze the factors influencing efficacy, and ROC curves was plotted to analyze the predictive value of RNFL and MRT alone or in combination for efficacy.RESULTS: RNFL and MRT of cases of severe amblyopia were higher than those of the mild to moderate amblyopia and the control groups(all P<0.05); the severity of amblyopia in children with refractive abnormalities is positively correlated with RNFL and MRT(rs=0.745 and0.724, both P<0.001); among patients of mild to moderate and severe, there were statistically significant differences between the effective and ineffective subgroups in terms of initial treatment age, fixation form, treatment compliance, as well as RNFL, MRT, and their differences before and 1mo postoperatively(all P<0.05). Logistic analysis showed that initial treatment age, fixation nature, treatment compliance, RNFL and MRT before and 1mo postoperatively were all factors influencing the therapeutic effect of amblyopia with refractive abnormalities in children(all P<0.05); after 1mo of treatment, the combined prediction of RNFL and MRT was significantly better than that of single prediction in children with mild to severe amblyopia.CONCLUSION:There are differences in RNFL and MRT in children with abnormal refractive amblyopia, and they are closely related to the different degrees and curative effects of children. The combination of RNFL and MRT after 1mo of treatment has certain value in predicting children with different degrees of abnormal refractive amblyopia.

Aim To explore the inhibitory effect of lenvatinib plus fluvastatin on liver transplantation tumor in mice and the mechanism.Methods Mouse model of subcutaneous liver cancer was used.Single agent of lenvatinib, single agent of fluvastatin, a combination of lenvatinib and fluvastatin and control solvent were given to four groups of mice.Tumor volume was measured.Immunohistochemistry was used to examine proliferation of tumor cells.Tunel was employed to detect the cell apoptosis.qRT-PCR and immunohistochemistry were used to measure the expression of TLR4.Western blot was employed to determine β-catenin expression.Rescue experiment was done using human hepatoma cells cultured in vitro.Results Treatment with both lenvatinib and fluvastatin significantly suppressed tumor growth in nude mice.Combined treatment significantly decreased the expressions of PNCA and increased apoptosis in tumor cells.Mechanically combined treatment synergistically suppressed the mRNA and protein expression of TLR4 which further inhibited the expression of β-catenin in hepatoma cells.Conclusions A combination of lenvatinib and fluvastatin synergistically inhibits tumor growth and promotes tumor cell apoptosis.The combination treatment significantly inhibits TLR4/β-catenin signaling pathway.

Objective To explore the recovery mechanism of language network among post-stroke aphasic patients by investigating the difference of whole-brain amplitude of low frequency fluctuations (ALFF) and the Granger causality analysis (GCA) between the patients and the controls. Methods From May, 2019 to May, 2021, 19 patients with aphasia after left hemispheric stroke and 17 age- and sex- matched healthy controls finished functional magnetic resonance imaging scanning. All the patients assessed with Chinese version of Western Aphasia Battery (WAB). Restplus was used for fMRI data analysis. Regions with significant difference of ALFF between groups were chosen as regions of interests (ROI) for the following GCA analysis. Results ALFF in left inferior frontal gyrus triangle (LIFGtri) and left medial frontal gyrus (LMFG) were significantly lower in the patients than in the controls. Effective connectivity from LIFGtri to LMFG and from LMFG to the right cerebellar Crus I were significantly lower in the patients. Effective connectivity from right cerebellar Crus II to LIFGtri, from right cerebellar Crus I to LMFG were significantly greater in the patients. Conclusion Unidirectional negative regulatory pathway such as LIFGtri→LMFG might be injured in post-stroke aphasia, while, connectivities between right cerebellar Crus II→LIFGtri and right cerebellar Crus I →LMFG enhanced. Right cerebellum might be the potential target for the language recovery.

Objective To investigate the effect of repetitive transcranial magnetic stimulation (rTMS) on the brain functional reorga-nization of aphasia after stroke, with functional magnetic resonance imaging (fMRI). Methods From January, 2017 to February, 2018, six eligible stroke patients with aphasia were recruited in experimental group, and nine age-gender matched healthy adults were recruited in healthy control group. Subjects in both groups received task-fMRI, and the experimental group was assessed with Chinese version of Western Aphasia Battery (WAB) examination before and after rTMS treatment. Four patients underwent rTMS at the right inferior frontal gyri pars triangularis marked by neuro-navigation-guided system, 1 Hz, five times per week for two weeks. The fMRI data were processed by SPM 12. The differences of brain activation and voxel changes be-tween two groups were compared. The fMRI data including the differences in brain activation, voxel volume and activation voxel indices (AVI) and WAB scores were analyzed before and after rTMS. Results The cerebral hemisphere activation in the experimental group was higher than that of the healthy control group, including the regions of interest (ROI) such as bilateral supplementary motor area and middle frontal gyrus, and the non-ROI (n-ROI) such as left praecuneus, left postcentral gyrus, right hippocampus, right paracingulate cor-tex, etc., while the activation reduced in the areas of left pars triangularis and n-ROI such as left calcarine fissure cortex, left gyrus lingualis, the right anterior cingulate and the paracingulate cortex. Cases 1 and 2 had shorter course of disease, smaller lesion volume, and activation increased in bilateral cerebral hemispheres before treat-ment. AVI showed that their hemispheric dominance was right, and activation reduced in bilateral cerebral hemi-sphere after treatment, but the high-efficiency language function area of ROI, such as the left pars triangularis, turned from inactive to active, and the hemispheric dominance lateralized from right to left, with the improve-ment of language function. For the case 3 and case 4, the disease courses were longer, the lesions sizes were larg-er, and both cerebral hemisphere activations were reduced before treatment. AVI showed that the hemispheric dominance of case 3 was right and was left in case 4. After treatment, bilateral cerebral hemispheres were activat-ed more than before, and the hemispheric dominance of language function was in the right hemisphere; the left middle frontal gyrus and right middle temporal gyrus were activated from no activation before treatment in case 3. The activation of the supplemental motor area on the right side was increased. In case 4, there was no activa-tion in ROI before treatment. After treatment, the bilateral supplementary motor area, right pars opercularis, and the right middle temporal gyrus were activated. Conclusion Low-frequency rTMS could improve the language function by optimizing bilateral cerebral hemisphere brain areas related with language function in patients with aphasia after stroke.

ObjectiveTo investigate the value of micro- dissection testicular sperm extraction (micro-TESE) in the treatment of non-obstructive azoospermia (NOA) in patients with the history of secondary testicular injury.

METHODSTotally, 121 NOA patients with the history of secondary testicular injury underwent micro-TESE in our hospital from September 2014 to December 2017. We analyzed the correlation of the sperm retrieval rate with the causes of testicular injury and compared the outcomes of the ICSI cycles with the sperm retrieved from the NOA males by micro-TESE (the micro-TESE group) and those with the sperm ejaculated from severe oligospermia patients (sperm concentration <1×10⁶/ml, the ejaculate group). Comparisons were also made between the two groups in the female age, two-pronucleus (2PN) fertilization rate, transferrable embryos on day 3 (D3), D3 high- quality embryos, D14 blood HCG positive rate, embryo implantation rate, and clinical pregnancy rate.

RESULTSTesticular sperm were successfully retrieved by micro-TESE in 86.0% of the patients (104/121), of whom 98.4% had the history of orchitis, 75.5% had been treated surgically for cryptorchidism, and 63.6% had received chemo- or radiotherapy. No statistically significant differences were observed between the micro-TESE and ejaculate groups in the 2PN fertilization rate (59.4% vs 69.3%, P > 0.05), D14 blood HCG positive rate (44.6% vs 57.9%, P > 0.05), embryo implantation rate (31.8 %% vs 32.6%, P > 0.05) and clinical pregnancy rate (41.5% vs 48.7%, P > 0.05). However, the rate D3 transferrable embryos was significantly lower in the micro-TESE than in the ejaculate group (40.5% vs 52.2%,P < 0.05), and so was that of D3 high-quality embryos (32.5% vs 42.1%, P < 0.05).

CONCLUSIONSMicro-TESE can be applied as the first choice for NOA patients with the history of secondary testicular injury, but more effective strategies are to be explored for the improvement of ICSI outcomes with the sperm retrieved by micro- TESE.

Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.

In this paper, the potential distribution information and ecological suitability regionalization for Gastrodia elata in Zhaotong were studied based on the climate, terrain, soil and vegetation factors analysis by Maxent and ArcGIS. The results showed that the highly potential distribution (suitability index>0.6) mainly located in Zhaotong, Yunnan province（Zhenxiong,Yiliang and Daguan county, with an area of 2 872 km²）, and Bijie, Guizhou province （Hezhang,Bijie,Weining county, 1 251 km²）. The AUC of ROC curve was above 0.99, indicating that the predictive results with the Maxent model were highly precise. The main ecological factors determining the potential distribution were the altitude, average rainfall in November, average rainfall in October, vegetation types, average rainfall in March, average rainfall in April,soil types,isothermal characteristic and average rainfall in June. The environmental variables in the highly potential areas were determined as altitude around 1 450-2 200 m,annual average temperature around 18.0-20.4 ℃,annual average precipitation around 900 mm,yellow soil or yellow brown soil,and acid sandy loam or slightly acidic sandy loam.The results will provide valuable references for plantation regionalization and the siting for imitation wild planting of G. elata in Zhaotong.