ObjectiveTo explore the material basis of the "interaction of blood stasis and toxin" mechanism in cerebral ischemia-reperfusion injury， as well as the protective role of Huoxue Huayu Jiedu prescription （HXHYJDF） against ferroptosis. MethodsSixty SPF-grade male SD rats were randomly divided into six groups： sham group， model group， deferoxamine （DFO） group （100 mg·kg^-1）， low-dose HXHYJDF group （4.52 g·kg^-1）， medium-dose HXHYJDF group （9.04 g·kg^-1）， and high-dose HXHYJDF group （18.07 g·kg^-1）， with ten rats in each group. Except for the sham group， the other groups were used to replicate the model of focal cerebral ischemia-reperfusion in the middle cerebral artery of rats by the reforming Longa method. Neurological function was assessed at 1^st， 3^rd， 5^th， and 7^th days post-reperfusion using the modified neurological severity scores （m-NSS）. Brain tissue pathology and the morphology of mitochondria were observed using hematoxylin-eosin （HE） staining and transmission electron microscopy. The contents of malondialdehyde （MDA）， glutathione （GSH）， divalent iron ions （Fe²⁺）， and reactive oxygen species （ROS） in the ischemic cerebral tissue were detected using enzyme-linked immunosorbent assay （ELISA）. Immunohistochemistry and Western blot （WB） were used to detect the expression of iron death marker proteins glutathione peroxidase 4 （GPX4）， ferroportin-1 （FPN1）， transferrin receptor protein 1 （TfR1）， and ferritin mitochondrial （FtMt） in brain tissue. ResultsCompared with the sham group， the mNSS score of the model group was significantly increased （P<0.01）. HE staining showed that the number of neurons in the cortex of brain tissue was seriously reduced， and the intercellular space was widened. The nucleus was fragmented， and the cytoplasm was vacuolated. The results of transmission electron microscopy showed that the mitochondria in the cytoplasm contracted and rounded， and the mitochondrial cristae decreased. The matrix was lost and vacuolated， and the density of the mitochondrial bilayer membrane increased. The results of ELISA showed that the content of GSH decreased significantly （P<0.01）， and the contents of MDA， Fe²⁺， and ROS increased significantly （P<0.01）. The results of immunohistochemistry and WB showed that the expression of GPX4 and FPN1 proteins was significantly decreased （P<0.01）， and the expression of FtMt and TfR1 proteins was significantly increased （P<0.01）. Compared with those of the model group， the m-NSS scores of the high-dose and medium-dose HXHYJDF groups began to decrease on the 3^rd and 5^th days， respectively （P<0.05， P<0.01）. The results of HE and transmission electron microscopy showed that the intervention of HXHYJDF improved the pathological changes of neurons and mitochondria. The results of ELISA showed that the content of GSH in the medium-dose and high-dose HXHYJDF groups increased significantly （P<0.01）， and the contents of MDA， Fe²⁺， and ROS decreased significantly （P<0.05， P<0.01）. The content of GSH in the low-dose HXHYJDF group increased significantly （P<0.01）， and the contents of MDA and ROS decreased significantly （P<0.01）. The results of immunohistochemistry showed that the expression of GPX4 and FPN1 in the high-dose HXHYJDF group increased significantly （P<0.01）， and the expression of FtMt and TfR1 decreased significantly （P<0.01）. The expression of GPX4 and FPN1 in the medium-dose HXHYJDF group increased significantly （P<0.05）， and the expression of TfR1 decreased significantly （P<0.01）. WB results showed that the expression levels of FPN1 and GPX4 proteins in the high-dose， medium-dose， and low-dose HXHYJDF groups were significantly up-regulated （P<0.01）， and the expression levels of FtMt and TfR1 proteins were significantly down-regulated （P<0.01）. ConclusionHXHYJDF can significantly improve neurological dysfunction symptoms in rats with cerebral ischemia-reperfusion injury， improve the pathological morphology of the infarcted brain tissue， and protect the brain tissue of rats with cerebral ischemia-reperfusion injury to a certain extent. Neuronal ferroptosis is involved in cerebral ischemia-reperfusion injury， with increased levels of MDA， Fe²⁺， ROS， and TfR1 and decreased levels of FtMt， FPN1， GPX4， and GSH potentially constituting the material basis of the interaction of blood stasis and toxin mechanism in cerebral ischemia-reperfusion injury. HXHYJDF may exert brain-protective effects by regulating iron metabolism-related proteins， promoting the discharge of free iron， reducing brain iron deposition， alleviating oxidative stress， and inhibiting ferroptosis.

Objective: To explore the differences in the gut microbiota of primary school students with different levels of sugar sweetened beverage intake, so as to provide scientific evidence for better identification of health risks in children and the development of targeted health policies. Methods: In June 2022, a total of 192 healthy primary school students from Chengdu were selected using a stratified cluster random sampling method. The sugar sweetened beverage intake was assessed through a dietary frequency questionnaire. Based on the median daily sugar sweetened beverage intake, primary school students were categorized into a low intake group ( n =96) and a high intake group ( n =96). The gut microbiota in fresh fecal samples from the two groups of primary school students was analyzed using 16S rRNA high throughput sequencing, and the diversity and community structure differences in the gut microbiota were compared. Results: Children in the low intake group had a sugar sweetened beverage intake of (21.3±1.6) mL/d, while the high intake group had an intake of (269.6±37.3) mL/d. Diversity analysis results showed that there were no statistically significant differences between the low intake and the high intake group in terms of α diversity metrics: Observed_otus index [298.50 (259.75, 342.25), 305.50 (244.25, 367.75)], Goods_coverage index [1.00 (1.00, 1.00), 1.00 (1.00, 1.00)], Chao index [304.18 (260.75, 348.78), 305.88 (245.68, 370.88)], Shannon index [5.88 (5.29, 6.45), 5.71 (4.89, 6.28)] and Simpson index [0.95 (0.91, 0.97), 0.94 (0.88, 0.97)] ( Z =-0.64, -0.76, -0.54, -1.76, -1.67, P >0.05). Furthermore, no statistically significant difference was observed in β diversity between the two groups ( R 2=0.006, P >0.05). At the genus level, the abundance of Blautia [0.033 (0.018, 0.055)] and Fusicatenibacter [0.009 (0.005, 0.015)] were higher in the low intake group compared to the high intake group [0.024 (0.013, 0.041),0.006 (0.003, 0.011)]and differences were statistically significant ( Z =-2.52, -2.81, P <0.05). LEfSe analysis highlighted intergroup differences primarily in Blautia, Fusicatenibacter and Sarcina( LDA= 3.56,3.12,3.53, P <0.05). Conclusions There is no significant difference in the diversity and overall structure of the gut microbiota in primary school students with different levels of sugar sweetened beverage intake. However, there are species variations at the genus level. The information can serve as a scientific basis for identifying health risks in primary school students and formulating targeted health strategies.

Objective: To determine the association between the intake of five major types of prepackaged foods and the growth and development of school aged children, so as to provide a theoretical basis for guiding school aged children and their parents to make healthy prepackaged food choices. Methods: Based on data from the South West China Childhood Nutrition and Growth Cohort (SCCNG), 381 children (6-11 years of age) were selected by stratified cluster sampling. Dietary intake and pubertal development were collected using questionnaires, and anthropometric measurements were obtained. Children were followed up until November 2022. Binary Logistic regression models were used to analyze the prospective association between prepackaged food intake and the growth and development of school aged children. Results: The total intake of the five major types of prepackaged foods was 316.1 (197.1,501.4) g/d. After 2 years of follow up evaluations, 16.5% of school aged children were shown to be overweight and obese. Early spermarche occurred in 12.6% of boys and early menarche occurred in 15.4% of girls. The following findings were suggested after adjusting for the mothers education level, average gross monthly family income, whether or not the family had one child only, geographic area of residence, body mass index Z score, average duration of daily exercise, and total dietary energy intake: convenience food intake might increase the risk of early spermarche ( OR =9.37); fruit and vegetable intake might decrease the risk of early spermarche and menarche ( OR =0.33,0.17); and fish, poultry, meat, and egg intake might increase the risk of early menarche ( OR =7.59)( P <0.05). Intake of the five types of prepackaged foods was not associated with being overweight or obese after adjusting for confounders ( OR =1.40, 0.57, 0.73, 1.33,1.57, P >0.05). Conclusions The relationship between intake of the five major types of prepackaged foods and pubertal development is inconsistent and no significant correlation was detected between the intake of prepackaged foods and overweight or obese children. Nutrition education should be strengthened to help children and their parents choose healthy prepackaged foods.

The contents, application progress, application effect and optimization strategy of group pregnancy health care model were reviewed, in order to provide reference for the establishment of standardized intervention and health management practice strategies of rural women′s pregnancy care in line with China′s national conditions.

Rabies is a zoonotic disease that poses a global public health threat.Rabies virus(RABV)is neurotropic and can cause severe neurological disorders and behavioral abnormalities in host,with a fatality rate nearly 100％.In order to identify the key genes for RABV affecting neuronal cell function,we established and analyzed the mRNA expression profile of Neuro-2a(N2a)cell in-fected with challenge virus standard(CVS)-11 by RNA sequencing(RNA-seq).Biological func-tions of differentially expressed genes(DEGs)were determined by GO and KEGG enrichment a-nalysis.The results showed that there were 415 differentially expressed genes in N2a infected with CVS-11 strain,of which 89 were up-regulated and 326 were down-regulated.These genes were re-lated to a variety of biological processes,such as axon guidance pathway,cholesterol metabolism pathway,nitrogen metabolism pathway,and glycosphingolipid biosynthesis pathway,many of them have been shown to be closely associated with RABV infection.A total of 12 DEGs related to axon conduction,antigen processing and presentation pathways were selected and detected by real-time PCR,and their expression trends were consistent with the RNA-seq results.The genomic tran-scriptomic data on N2a cell under RABV infection will provide new clues for probing the mecha-nisms of RABV infection and transmission in the nervous system in the future.

Objective To explore the value of synchrotron radiation micro-CT(SR-pCT)and automatic marker watershed image segmentation for quantifying three-dimensional local bone tissue and blood vessels in mouse models of breast cancer tibia metastasis.Methods Mouse models with breast cancer bone metastasis in tibia were established and underwent oral Vatalanib,injection of Bevacizumab,combination of Vatalanib or Bevacizumab,or injection of placebo,respectively.On the 8th day(time point T1)and the 22nd day(time point T2)after injection of tumor cells,the tibia bone was scanned using combined vascular contrast injection and SR-pCT,and high-resolution simultaneous imaging of local bone tissue and vascular network were achieved.Then bone tissue and vessels on images were segmented using automatic marker watershed image-segmentation method,global threshold method and region growing method,respectively,and the quality of segmentations were assessed with Dice similarity coefficient(DSC).The anatomical quantitative parameters of tibia with breast cancer metastasis were obtained,including the fraction of bone volume in total volume(BV/TV),fraction of vessel volume in total volume(VV/TV),the mean thickness of bone(B.Th)and the mean thickness of vessel(V.Th).The anatomical quantitative parameters were compared among different treatments at the same time point,also between different time points under the same treatment.Results DSC for segmentation of local tibia bone tissue and vascular structures based on automatic marker watershed image segmentation method were all greater than 0.95,higher than that of other methods(all P＜0.05).No significant difference of anatomical quantitative parameters was found under different treatments at T1(all P＞0.05),and V.Th under the combined treatment were lower than that under other treatments at T2(all P＜0.05).Compared with T1,BV/TV and B.Th at T2 increased while VV/TV at T2 decreased in tibia metastasis under combination treatment(all P＜0.05),BV/TV,B.Th and V.Th at T2 increased under Vatalanib or Bevacizumab treatment(all P＜0.05),and BV/TV and B.Th at T2 increased under placebo(both P＜0.05).Conclusion The combination of SR-pCT and automatic marker watershed method could be used to initially quantify three-dimensional structures of local bone tissue and blood vessels in mouse models of breast cancer tibia metastasis.

Objective:To evaluate the reliability and validity of the simplified Chinese version of Lymphedema function, disability and health questionnaire for lower limb lymphedema (Lymph-ICF-LL).Methods:The original English version was translated into mandarin and modified to accommodate Chinese customs. According to the inclusion and exclusion criteria, 61 patients with lower extremity lymphedema who were treated in the Department of Burn and Plastic Surgery of Xi’an Central Hospital from June 2019 to June 2021 were recruited in this study. The questionnaires filled by the included patients were classified as sample 1 to sample 4 by the screening process. The valid simplified Chinese version of Lymph-ICF-LL was collected as sample 1 ( n=55), including 1 male and 54 females, aged (51.0±9.4) years (24-70 years). Among the 55 patients, 28 patients underwent lymphaticovenular anastomosis (LVA). The simplified Chinese version of Lymph-ICF-LL completed before and follow-up of 6 months after surgery were selected as sample 2 and sample 3( n=28), respectively. The simplified Chinese version of lymphoedema quality of life questionnaire (LYMQOL) filled out by the patients in sample 1 was used as sample 4 ( n=55). The reliability was analyzed by internal consistency test reliability and split-half reliability, Cronbach’s α coefficient ≥0.80 indicated good internal consistency reliability and the Spearman-Brown split-half reliability coefficient ≥0.70 indicated good split-half reliability. The content validity of the Lymph-ICF-LL was determined by three plastic surgery experts who judged the consistency of the questionnaire with the original English content. Exploratory factor analysis and criterion validity were used to analyze construct validity. For criterion validity assessment, Pearson correlation analysis was used to test the correlation between the Lymph-ICF-LL scores in sample 1 and the LYMQOL scores in sample 4. The scores of Lymph-ICF-LL before and half a year after lymphadenectomy were compared by paired t-test. Results:(1) Cronbach’s alpha was 0.937, indicating good internal consistency. The Spearman-Brown split-half reliability coefficient of the total scale was 0.700, indicating good split-half reliability. (2) The content validity of the simplified Chinese version of Lymph-ICF-LL was good. Five factors (physical function, mental function, general tasks/household, mobility and life/social life) were identified by factor analysis. The total score of the simplified Chinese version of Lymph-ICF-LL was positively correlated with the score of the simplified Chinese version of LYMQOL ( r=0.73, P<0.001), and the criterion correlation validity was good. (3) The scores of Lymph-ICF-LL for half a year after LVA was significantly lower than its preoperative counterparts (40.70±15.13 vs. 54.40±15.70, t=5.96, P<0.001). The quality of life of patients after LVA was significantly improved. Conclusion:The simplified Chinese version of Lymph-ICF-LL shows good reliability and validity and can be used to evaluate the quality of daily life regarding patients with lower limb lymphedema.

Objective:To evaluate the reliability and validity of the simplified Chinese version of Lymphedema function, disability and health questionnaire for lower limb lymphedema (Lymph-ICF-LL).Methods:The original English version was translated into mandarin and modified to accommodate Chinese customs. According to the inclusion and exclusion criteria, 61 patients with lower extremity lymphedema who were treated in the Department of Burn and Plastic Surgery of Xi’an Central Hospital from June 2019 to June 2021 were recruited in this study. The questionnaires filled by the included patients were classified as sample 1 to sample 4 by the screening process. The valid simplified Chinese version of Lymph-ICF-LL was collected as sample 1 ( n=55), including 1 male and 54 females, aged (51.0±9.4) years (24-70 years). Among the 55 patients, 28 patients underwent lymphaticovenular anastomosis (LVA). The simplified Chinese version of Lymph-ICF-LL completed before and follow-up of 6 months after surgery were selected as sample 2 and sample 3( n=28), respectively. The simplified Chinese version of lymphoedema quality of life questionnaire (LYMQOL) filled out by the patients in sample 1 was used as sample 4 ( n=55). The reliability was analyzed by internal consistency test reliability and split-half reliability, Cronbach’s α coefficient ≥0.80 indicated good internal consistency reliability and the Spearman-Brown split-half reliability coefficient ≥0.70 indicated good split-half reliability. The content validity of the Lymph-ICF-LL was determined by three plastic surgery experts who judged the consistency of the questionnaire with the original English content. Exploratory factor analysis and criterion validity were used to analyze construct validity. For criterion validity assessment, Pearson correlation analysis was used to test the correlation between the Lymph-ICF-LL scores in sample 1 and the LYMQOL scores in sample 4. The scores of Lymph-ICF-LL before and half a year after lymphadenectomy were compared by paired t-test. Results:(1) Cronbach’s alpha was 0.937, indicating good internal consistency. The Spearman-Brown split-half reliability coefficient of the total scale was 0.700, indicating good split-half reliability. (2) The content validity of the simplified Chinese version of Lymph-ICF-LL was good. Five factors (physical function, mental function, general tasks/household, mobility and life/social life) were identified by factor analysis. The total score of the simplified Chinese version of Lymph-ICF-LL was positively correlated with the score of the simplified Chinese version of LYMQOL ( r=0.73, P<0.001), and the criterion correlation validity was good. (3) The scores of Lymph-ICF-LL for half a year after LVA was significantly lower than its preoperative counterparts (40.70±15.13 vs. 54.40±15.70, t=5.96, P<0.001). The quality of life of patients after LVA was significantly improved. Conclusion:The simplified Chinese version of Lymph-ICF-LL shows good reliability and validity and can be used to evaluate the quality of daily life regarding patients with lower limb lymphedema.

Objective: To investigate the rationale for appropriate diagnostic methods and treatment protocols for unexpected gallbladder carcinoma(UGC). Methods: The clinical and pathological data of 45 patients with UGC admitted at Department of General Surgery, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine,from January 2008 to December 2017 were retrospectively collected and analyzed.There were 11 males(28.9%) and 34 females(71.1%),aged 68 years(range:27 to 68 years).And there were 20 cases who aged above 70 years. Twenty-four cases were diagnosed preoperatively as cholecystolithiasis plus chronic cholecystitis.Ten cases were diagnosed preoperatively as cholecystolithiasis plus actue cholecystitis.Six cases were diagnosed preoperatively as cholecystolithiasis plus choledocholith.Six cases were admitted because of gallbladder polyp and 1 case was admitted because of gallbladder adenomyomatosis. Results: Thirty-four patients with UGC received radical surgery.Among them,11 patients experienced postoperative complication and no posterative mortality occoured during hospital stay.Thirteen patients were diagnosed with T1b UGC, the harvested lymph node of Nx, N0, N1 and N2 was 2, 9, 1 and 1, respectively.In addition, 2 cases were identified to have local-regional tumor recurrence during our rescue radical surgery.The median overall survival time of the patients who did not receive radical surgery was 7 months(range:2-56 months).Nevertheless,the median overall survival time for patients diagnosed with T1, T2 and T3 tumors who received radical surgery, was 41 months(range: 19-82 months), 33.5 months(range: 31-36 months) and 17 months(range: 7-46 months), respectively. Conclusions For patients with UGC, rescue radical surgery can achieve a better survival time.Furhtermore, our experience proved that rescue radical surgery for UGC is safe and feasible.Therefore,rescue radical surgery should be performed in patients with diagnose with UGC especially those T1b patients.