ObjectiveTo explore the effect of Buzhong Yiqitang on exercise-induced fatigue and its potential mechanism. MethodsSixty male SPF-grade C57BL/6J mice were randomized into blank， model， low-， medium-， high-dose （4.1， 8.2， 16.4 g·kg^-1， respectively） Buzhong Yiqitang， and vitamin C （0.04 g·kg^-1） groups. The blank and model groups were administrated with normal saline. Each group was administrated with corresponding agents by gavage at a dose of 0.2 mL once a day. Except the blank group， other groups underwent a 6-weeks exhaustive swimming test under negative gravity. At the end of the experiment， blood was collected， and the thymus， spleen， liver， and kidney weights were measured. Serum levels of lactic acid （LD）， blood urea nitrogen （BUN）， creatine kinase （CK）， and malondialdehyde （MDA） were assessed by kits to evaluate fatigue. Hematoxylin-eosin staining was performed to observe pathological changes in the skeletal muscle. Electron microscopy was used to examine the skeletal muscle cell ultrastructure， with a focus on mitochondrial morphological changes. The adenosine triphosphate （ATP） content and activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， and Ⅴ in skeletal muscle were determined by kits. The expression levels of key genes and proteins in the PTEN-induced putative kinase 1 （PINK1）-mediated mitochondrial homeostasis pathways in the skeletal muscle were evaluated via Real-time PCR and Western blot， respectively. ResultsCompared with the blank group， the model group showed reductions in weight gain rate （P<0.01） and thymus index （P<0.01）， rises in serum levels of LD， BUN， MDA， and CK （P<0.01）， disarrangement of skeletal muscle， broken muscle fibers， inflammatory cell infiltration in muscle fiber gaps， abnormal morphological changes （increased vacuolated mitochondria and disappearance of cristae） of mitochondria in skeletal muscle cells， and decreased mitochondria. In addition， the skeletal muscle in the model group showed reduced content of ATP， weakened activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， and Ⅴ （P<0.05）， up-regulated mRNA levels of PINK1， E3 ubiquitin-protein ligase （Parkin）， hairy/enhancer-of-split related with YRPW motif 1 （HEY1）， dynamin-related protein 1 （Drp1）， sequestosome 1 （p62）， and hypoxia-inducible factor 1 alpha （HIF-1α） （P<0.05）， and down-regulated protein level of microtubule-associated protein 1-light chain 3B （LC3B） （P<0.01）. Compared with the model group， Buzhong Yiqitang prolonged the swimming exhaustion time （P<0.01）， increased the weight gain rate （P<0.01） and thymus index （P<0.01）， lowered the serum levels of LD， BUN， MDA， and CK （P<0.05， P<0.01）. The skeletal muscle in the Buzhong Yiqitang groups showed neat arrangement， reduced inflammatory cells， intact mitochondria with dense cristae， and increased mitochondria. In addition， the skeletal muscle in the Buzhong Yiqitang groups showcased increased ATP content， enhanced activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， and Ⅴ （P<0.05， P<0.01）， up-regulated protein levels of PINK1， Parkin， HEY1， LC3B， and Drp1 and mRNA level of HIF-1α （P<0.05， P<0.01）， and down-regulated expression level of p62 （P<0.05， P<0.01）. ConclusionBuzhong Yiqitang can prevent and treat exercise-induced fatigue by regulating the mitochondrial homeostasis of skeletal muscle via the HIF-1α/PINK1/Parkin and HIF-1α/HEY1/PINK1 signaling pathways.

ObjectiveTo explore the effect of Buzhong Yiqitang on exercise-induced fatigue and its potential mechanism. MethodsSixty male SPF-grade C57BL/6J mice were randomized into blank， model， low-， medium-， high-dose （4.1， 8.2， 16.4 g·kg^-1， respectively） Buzhong Yiqitang， and vitamin C （0.04 g·kg^-1） groups. The blank and model groups were administrated with normal saline. Each group was administrated with corresponding agents by gavage at a dose of 0.2 mL once a day. Except the blank group， other groups underwent a 6-weeks exhaustive swimming test under negative gravity. At the end of the experiment， blood was collected， and the thymus， spleen， liver， and kidney weights were measured. Serum levels of lactic acid （LD）， blood urea nitrogen （BUN）， creatine kinase （CK）， and malondialdehyde （MDA） were assessed by kits to evaluate fatigue. Hematoxylin-eosin staining was performed to observe pathological changes in the skeletal muscle. Electron microscopy was used to examine the skeletal muscle cell ultrastructure， with a focus on mitochondrial morphological changes. The adenosine triphosphate （ATP） content and activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， and Ⅴ in skeletal muscle were determined by kits. The expression levels of key genes and proteins in the PTEN-induced putative kinase 1 （PINK1）-mediated mitochondrial homeostasis pathways in the skeletal muscle were evaluated via Real-time PCR and Western blot， respectively. ResultsCompared with the blank group， the model group showed reductions in weight gain rate （P<0.01） and thymus index （P<0.01）， rises in serum levels of LD， BUN， MDA， and CK （P<0.01）， disarrangement of skeletal muscle， broken muscle fibers， inflammatory cell infiltration in muscle fiber gaps， abnormal morphological changes （increased vacuolated mitochondria and disappearance of cristae） of mitochondria in skeletal muscle cells， and decreased mitochondria. In addition， the skeletal muscle in the model group showed reduced content of ATP， weakened activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， and Ⅴ （P<0.05）， up-regulated mRNA levels of PINK1， E3 ubiquitin-protein ligase （Parkin）， hairy/enhancer-of-split related with YRPW motif 1 （HEY1）， dynamin-related protein 1 （Drp1）， sequestosome 1 （p62）， and hypoxia-inducible factor 1 alpha （HIF-1α） （P<0.05）， and down-regulated protein level of microtubule-associated protein 1-light chain 3B （LC3B） （P<0.01）. Compared with the model group， Buzhong Yiqitang prolonged the swimming exhaustion time （P<0.01）， increased the weight gain rate （P<0.01） and thymus index （P<0.01）， lowered the serum levels of LD， BUN， MDA， and CK （P<0.05， P<0.01）. The skeletal muscle in the Buzhong Yiqitang groups showed neat arrangement， reduced inflammatory cells， intact mitochondria with dense cristae， and increased mitochondria. In addition， the skeletal muscle in the Buzhong Yiqitang groups showcased increased ATP content， enhanced activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， and Ⅴ （P<0.05， P<0.01）， up-regulated protein levels of PINK1， Parkin， HEY1， LC3B， and Drp1 and mRNA level of HIF-1α （P<0.05， P<0.01）， and down-regulated expression level of p62 （P<0.05， P<0.01）. ConclusionBuzhong Yiqitang can prevent and treat exercise-induced fatigue by regulating the mitochondrial homeostasis of skeletal muscle via the HIF-1α/PINK1/Parkin and HIF-1α/HEY1/PINK1 signaling pathways.

OBJECTIVE To explore the mechanism of improvement effect of Chanbao zhichuang suppository （CBZCS） on hemorrhoids in rats through network pharmacology and metabolomics. METHODS A hemorrhoid model was established by subcutaneous injection of rhododendron oil to induce anal swelling. SD rats were divided into blank group （NC group， 0.32 g/kg vaseline）， model group （Model group， 0.32 g/kg vaseline）， CBZCS low-， medium-， and high-dose groups （CBZCS-L， CBZCS- M， CBZCS-H groups， with dosages of 0.16， 0.32， and 0.64 g/kg respectively）， and Mayinglong musk hemorrhoids suppository group （Positive group， 0.32 g/kg）， with 9 rats in each group. Anal administration was performed at 6， 12， 24， 48， and 72 hours after modeling. After the last administration， the pathological changes of the anal tissues in rats were observed， and the serum levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in rats were detected. Differential metabolite analysis and enrichment analysis were conducted by metabolomics methods， and the target proteins of CBZCS in treating hemorrhoids were obtained by network pharmacology. The core metabolic pathways were screened by interaction and enrichment analysis of differential metabolites and proteins， and the core proteins were experimentally verified. RESULTS Compared with the NC group， the anal tissues of the Model group showed obvious lesions， and the levels of IL-6 and TNF- α in the serum were significantly increased （P＜0.05）； compared with the Model group， the pathological damage of the anal tissues in the treatment groups was alleviated to varying degrees， and serum levels of IL-6 in CBZCS-H group， CBZCS-M group， and Positive group as well as serum levels of TNF-α in CBZCS-H group were significantly reduced （P＜0.05）. The metabolomics results showed that 34 differential metabolites were screened from the anal tissues of rats， and 22 of them showed a return after CBZCS administration. The differential metabolites mainly enriched in arachidonic acid metabolism， histidine metabolism， and glycerophospholipid metabolism. Through the network pharmacology， 138 intersection genes of CBZCS against hemorrhoids were determined. The analysis results showed that differential metabolites and target proteins were mainly enriched in the arachidonic acid metabolism pathway， and the regulation of this pathway might be related to cyclooxygenase-2 （COX-2）， Myc proto-oncogene protein （c-MYC）， cytochrome P450 1B1 （CYP1B1）， interleukin-1β （IL-1β）， and IL-6 protein expression. The experimental verification results showed that the expression levels of key proteins （COX-2， c-MYC， CYP1B1， IL-6， IL-1β） in the anal tissues of the Model group were significantly higher than those in the NC group （P＜0.05）， and the levels of the above proteins in the anal tissues of CBZCS-H group and Positive group were significantly lower than those in the Model group （P＜0.05）. CONCLUSIONS The mechanism of CBZCS in treating hemorrhoids may be to inhibit the expression of COX-2， c-MYC and CYP1B1 proteins， thereby inhibiting arachidonic acid metabolism and reducing the release of inflammatory factors IL-6 and IL-1β.

Objective:To explore the assessment value of multimodal imaging technique for the hepatic fibrosis(HF)of rats.Methods:Forty-eight SD rats were selected,and they were divided into control group,M1 model group,M2 model group and M3 model group according to random number table,with 12 rats in each group.The three model groups were induced by combination method of carbon tetrachloride(CCl4)plus ethanol to establish HF model.At the 4th,6th and 8th week of modelling,the M1 model group underwent ultrasound examination,and the M2 model group underwent examination of magnetic resonance,and the M3 model group adopted the examinations of conventional ultrasound,ultrasound elastography and dynamically contrast enhanced magnetic resonance imaging(DCE-MRI).The rats of control group were randomly divided into three subgroups,with four rats in each subgroup,which were controlled with the rats in the M1,M2 and M3 groups,respectively.At the end of the imaging examinations,the HE staining were conducted on the liver tissues so as to observe the HF degree of rats.The conventional ultrasound was used to observe the morphology of the rat liver,and to measure the inner diameter of the main trunk of the portal vein.colour Doppler ultrasound was used to measure the blood flow velocity of portal vein.Ultrasound elastography was used to observe the color distribution of liver parenchymal stiffness,and to measure the elasticity value of liver parenchyma.The initial area under curve(iAUC)of the receiver operating characteristic(ROC)was adopted to analyze the volume transit constant(Ktrans),rate constant(Kep)and extravascular extracellular space volume fraction(Ve)of the hemodynamic parameters in enhanced magnetic resonance examination.Results:The conventional ultrasound showed there was no abnormal changes in the livers of rats in control group,and the morphology of the liver of M1 group occurred changes.Ultrasound elastography showed that the elasticity values of the liver of rats in M1 group appeared upward trend with the aggravation of the HF degree than control group.Conventional MRI of group M2 showed that the liver morphology was normal in the control group,and the changes of the liver morphologies of rats in M2 group at the 6th and 8th week post modeling were more obvious than that at the 4th week post modeling.The ROC curve analysis of DCE-MRI results showed that Ktrans and Ve had higher diagnostic efficacy for HF,and their iAUC values were>0.9.The results of intergroup comparison of rats among M1 model group,M2 model group and M3 model group indicated that the sensitivity,accuracy,positive predictive value and negative predictive value of multimodal imaging technique were significantly higher than those of single examination in diagnosing HF at early stage of rats,and the differences were statistically significant(x2=0.634,0.644,0.621,0.543,P<0.05),respectively.HE staining results showed that the liver tissues of the rats in the model groups appeared fibroplasia of different degrees.Conclusion:Multimodal imaging technique can be used as an effective test method of evaluating hepatic fibrosis.

Objective To investigate the clinical characteristics of elderly patients with acute pancreatitis(AP),and to provide a basis for the prevention and treatment of elderly patients with AP.Methods Clinical data of AP patients treated in Jiangdu People's Hospital Affiliated to Yangzhou University from March 2021 to March 2023 were collected and divided into elderly group(≥65 years old,167 cases)and non-elderly group(＜65 years old,303 cases)according to age.The clinical characteristics,biochemical indexes,mortality,intensive care unit(ICU)hospitalization rate,length of stay,Ranson score,Charlson comorbidity index(CCI)and so on were compared between the two groups.Results The age of elderly group was significantly higher than that of non-elderly group,lactate dehydrogenase,aspartate aminotransferase,serum amylase,Ranson score,the proportion of cardiovascular complications,pulmonary complications,nervous system complications,CCI ≥ 2 and the incidence of jaundice were significantly higher than those of non-elderly group(P＜0.05).The mortality rate and ICU hospitalization rate of elderly group were significantly higher than that of non-elderly group,and the hospital stay was significantly longer than that of non-elderly group(P＜0.05).Multivariate analysis showed that age ≥65 years,Ranson score ≥3,CCI≥ 2 and pancreatic necrosis were independent risk factors for death and ICU admission in AP patients(P＜0.05),age ≥65 years,pancreatic necrosis,and biliary causes were independent risk factors for prolonged hospital stay(P＜0.05).Conclusion Compared with middle-aged and young patients,elderly AP patients have higher mortality,higher ICU hospitalization rate and longer hospital stay.Early identification and timely treatment are key factors to improve prognosis elderly AP patients.

Single-cell transcriptome sequencing (scRNA-seq) can resolve the expression characteristics of cells in tissues with single-cell precision, enabling researchers to quantify cellular heterogeneity within populations with higher resolution, revealing potentially heterogeneous cell populations and the dynamics of complex tissues. However, the presence of a large number of technical zeros in scRNA-seq data will have an impact on downstream analysis of cell clustering, differential genes, cell annotation, and pseudotime, hindering the discovery of meaningful biological signals. The main idea to solve this problem is to make use of the potential correlation between cells and genes, and to impute the technical zeros through the observed data. Based on this, this paper reviewed the basic methods of imputing technical zeros in the scRNA-seq data and discussed the advantages and disadvantages of the existing methods. Finally, recommendations and perspectives on the use and development of the method were provided.
Cluster Analysis ; Transcriptome

Objective:To study the effect of experiential teaching combined with literature salon on the teaching quality of neurosurgery standardized training.Methods:A total of 120 trainees having standardized training in the Department of Neurosurgery of Southwest Hospital, Army Medical University from January 2018 to January 2020 were selected in the study, and they were divided into study group ( n=67) and control group ( n=53) according to the time of admission. The control group received traditional clinical teaching, and the study group was given experiential teaching combined with literature salon. The assessment results (scores of theoretical examination, clinical practice skill and case analysis skill), problem analysis skills and self-confidence of critical thinking were compared between the two groups before and after teaching, and the teaching satisfaction of the two groups was compared. SPSS 20.0 was performed for t test and chi-square test. Results:After teaching, the theoretical examination score, clinical practice skill score and case analysis ability score of the two groups were significantly increased compared with those before teaching, and the clinical practice skill score and case analysis ability score of study group were significantly higher than those of control group ( P<0.05). After teaching, the scores of problem analysis ability and self-confidence of critical thinking of the two groups were significantly higher than those before teaching, and the scores of problem analysis ability and self-confidence of critical thinking were significantly higher in study group than those in control group ( P<0.05). The teaching satisfaction of trainees in study group (91.04%, 61/67) was significantly higher than that in control group (73.58%, 39/53) ( P<0.05). Conclusion:Experiential teaching combined with literature salon teaching can significantly improve the quality of clinical teaching in department of neurosurgery, and it is helpful to improve the clinical practical ability and enhance the teaching satisfaction of students with standardized training.

Objective:To investigate the implementation procedures and dosimetric verification of the first patient treated with total body irradiation (TBI) based on volumetric modulated arc therapy (VMAT).Methods:Two sets of CT images were acquired under the head-in first and foot-in first to contour the planning target volume (PTV) of the cranial and caudal segments to accomplish the treatment of the whole body length, on which two interrelated plans of 5 subsequent isocenters with a total of 15 VMAT fields were performed to cover all PTVs. The plans were prescribed to ensure 90% PTV dose coverage with a total dose of 12 Gy in 6 fractions. Firstly, a dose optimization was performed on the caudal CT images, then the cranial CT images were optimized based on the dose distribution of the caudal CT images. The evaluation of the final treatment plan was carried out based on a plan sum of both two sets of images. The parameters of PTV and organs at risk (OARs) were measured by dose volume histograms from the accumulated plan. The quality assurance comprised the verification of the VMAT plans for each individual isocenter via Delta4 phantom. The dose distribution in the overlapped region between two adjacent central fields was verified with EBT3 film. The absolute dose at the overlapped region between two images was measured via Pinpoint chamber. In vivo dosimetry on the patient′s skin was monitored by MOSFET dosimeters. The results of planning parameters and treatment duration were analyzed. Results:The mean doses of two segments of PTVs were 12.45 Gy and 12.37 Gy. The mean dose for the lung was 10.8 Gy. The machine unit (MU) and mean treatment delivery time were 2 883 MU and 24.3 min, and the mean total time per fraction was 121 min. The mean 3%/3 mmγ-analysis pass rate for each isocenter VMAT plan was (99.74±0.42)%, and the mean 5%/5 mmγ-analysis pass rate for the overlapped region was (90.11±2.72)%. The average deviation of absolute dose in the overlap region of the caudal and cranial images was (3.6±0.4)%. In vivo measurement of 8 points on the patient showed that the dose of each region was ranged from 1.57 Gy to 2.04 Gy. Conclusion:According to the results of dosimetric verification, TBI based on multi-isocenter VMAT can be applied in clinical practice, which remains to be improved in terms of dose distribution, measurement results and clinical efficiency.

Objective:To analyze the relationship of hepatic transit time and portal vein pressure in the patients with portal hypertension and esophagogastric varices, and to clarify its clinical significance.Methods:A total of50hospitalized patients underwent esophagogastric venous embolization due to portal hypertension and esophagogastric varices were selected as observation group, and 50patients without history of liver disease and clinical manifestations of liver disease (hospitalized or out-patient) were selected as control group, All patients underwent liver contrast enhanced ultrasound.The free portal pressure (FPP) was measured by ultrasound guided portal vein puncture for operating pathways in the patients in observation group.The hepatic artery-hepatic vein transit time (HA-HVTT) , the hepatic portal vein-hepatic vein transit time (PV-HVTT) , the liver parenchymahepatic vein transit time (PA-HVTT) of the patients were compared between two groups.The correlations between HA-HVTT, PV-HVTT, PA-HVTT and FPP of the patients in observation group were analyzed.Results:The HA-HVTT, PV-HVTT and PA-HVTT of the patients in observation group were significantly shorter than those in control group (t=5.078, P＜0.01;t=12.163, P＜0.01;t=2.649, P＜0.01) .The HA-HVTT, PV-HVTT and PA-HVTT had negative correlations with FPP in observation group (r=-0.799, P＜0.01;r=-0.554, P＜0.01;r=-0.735, P＜0.01) .The linear relationship between HA-HVTT and FPP was Y=-0.410X+7.254;the linear relationship between PV-HVTT and FPP was Y=-0.355X+4.983;the linear relationship between PA-HVTT and FPP was Y=-0.566X+4.997.Conclusion:Liver transit time can be used as an effective index to judge the portal vein pressure, and it can provide the theroretical basis for the diagnosis and treatment of portal hypertension in the patients.

Objective: To investigate the expression and function of the TNF signaling pathway in the early stage of orthodontic tooth movement with periodontitis and to provide evidence to study the early inflammatory response in patients with periodontitis orthodontic treatment. Methods: Sixteen SD rats were randomly divided into four groups: group A--12 h of orthodontic tooth movement of the bilateral maxillary first molars in rats with periodontitis; group B--periodontitis model of the bilateral maxillary first molars without orthodontic tooth movement; group C--12 h of orthodontic tooth movement of the same teeth in rats with healthy periodontium; group D--control group without operations. The bilateral maxillary first molars and surrounding periodontal tissue of each group were collected for gene chip detection. Pathway enrichment analysis, qRT-PCR and GO (gene ontology) analysis were performed to identify differential genes involved in the TNF signaling pathway. Results : Gene chip results showed that the TNF signaling pathway was significantly upregulated in group A, group B and group C (P <0.01). Among the differential genes involved in the pathway, 28 were upregulated and 5 were downregulated in group A, 12 were upregulated and 4 were downregulated in group B, and 12 were upregulated and 1 was downregulated in group C (P <0.05). The most significant GO items included "response to lipopolysaccharide", "inflammatory response", "positive regulation of NF-κB transcription factor activity", "positive regulation of NF-κB import into nucleus" and "response to hypoxia"(P <0.001). qRT-PCR results showed no significant difference in TNF-α mRNA expression in group C compared with that in group D, TNF-α was upregulated in both groups A and B (P <0.01), and mRNA expression decreased in the following order: group A > group B > group C (P <0.05). Compared with group D, the expression levels of prostaglandin-endoperoxide synthase 2 (PTGS2) and interleukin-6 ( IL-6) in groups A, B and C were significantly upregulated (P <0.05), but the expression levels of PTGS2 and IL-6 in group A were lower than those in group B (P < 0.05). Conclusion The TNF signaling pathway is activated in the early stage of orthodontic tooth movement in rats with periodontitis. The pathway products participate in many biological processes and play an important role in the inflammatory response and bone absorption.