Objective To explore the anti-depression mechanism of Kai-Xin-San(KXS)via regulation of neurogenesis in hippocampus of depression-like mice.Methods The extracts of KXS were prepared and the anti-depression effects of KXS were evaluated by behavioral tests on chronic unpredictable mild stress(CUMS)induced depression-like mice.Evaluating depression-like behavior in CUMS mice through sucrose preference test,forced swimming test,tail suspension test,and other methods.Neurogenesis in hippocampus were determined by immunofluorescence assay.In addition,effects of KXS on regulating nestin expression and Wnt/b-catenin signaling pathway were explored by western blotting analysis.Amounts of cortisol,corticotropin-releasing factor(CRF),adrenocorticotropic hormone(ACTH),brain-derived neurotrophic factor(BDNF)and nerve growth factor(NGF)were determined by ELISA tests.Mouse primary neural stem cells(NSC)was used to evaluate the effect of KXS on promoting its proliferation by immunofluorescence assay.In addition,effects of KXS on regulating nestin and Wnt/β-catenin signaling pathway were also explored by Western blotting analysis.Results KXS significantly ameliorated the depression-like behaviors in presence of increased sucrose preference rate and decreased immobile time of tail suspension and forced swimming.KXS significantly promoted the neurogenesis in the hippocampus and expressions of nestin,reduced the expressions of cortisol,CRF,ACTH,increased the expressions of BDNF,NGF,and regulated Wnt/β-catenin signaling pathway.KXS also promoted the proliferation of NSCs and expressions of nestin,enhanced the translocation of b-catenin into nucleus,and regulated the expressions of proteins of Wnt/β-catenin signaling pathway.Conclusion KXS promoted neurogenesis in hippocampus and regulated Wnt/β-catenin pathway,which might contribute to its antidepressant effect.

Objective Evaluation of the effect and mechanism research of Qi-Shen-Yi-Zhi formula on improving learning and memory ability in mice injected with Aβ1-42 in hippocampus.Methods Alzheimer's disease model mice were constructed by injecting β amyloid peptide 1-42 into hippocampus and treated with water extracts of Qi-Shen-Yi-Zhi formula.The cognitive abilities of mice were assessed using Morris water maze and Y maze tests,which measure learning and memory capabilities.HE staining was used to observe the damage and TUNEL method was used to determine apoptosis of hippocampus.Detection of the expression of oxidative factors,inflammatory factors,and related antioxidant proteins and apoptotic proteins in the hippocampal tissue of a mouse model of dementia.Results Both high-dose and low-dose groups of Qi-Shen-Yi-Zhi formula significantly improved cognitive dysfunction in mice injected with Aβ1-42 in hippocampus,and attenuated the damage and apoptosis of the hippocampus.It also inhibited oxidative stress and downregulated the expressions of inflammatory factors IL-6,IL-1β and TNF-a,increased the expression of antioxidant proteins Nrf2 and HO-1,and regulated the expressions of apoptotic proteins Caspase-9,Caspase-3,Bax and Bcl-2.Conclusion Qi-Shen-Yi-Zhi formula improves the learning and memory abilities of mice injected with Aβ1-42 in hippocampus,which might be related to the attenuation of oxidative stress and neuronal inflammation of hippocampus.

OBJECTIVE To evaluate the effect of Qishen Yizhi formula on improving learning and memory ability in D-galactose subcutaneous injection induced subacute aging mice.METHODS Subacute aging mice model mice were developed by D-galactose subcutaneous injection and then treated with positive drug donepezil(2 mg·kg-1·d-1)and Qishen Yizhi formula water extracts in low(1.33 g·kg-1·d-1)and high dose group(2.67 g·kg-1·d-1).The learning and memory abilities of mice were evaluated using Morris water maze and Y maze tests;HE staining was used to examine hippocampal damage in model mice;TUNEL was used to detect apoptosis of mouse hippocampal tissue;ELISA was used to detect the expression levels of oxidative stress factors and inflammatory fac-tors in the mouse hippocampus tissue;Western blot was used to detect the expression of signaling pathway proteins related to apoptosis,oxidative stress and inflammatory stress in the hippocampus of mice.RESULTS The water extract of Qishen Yizhi formula signifi-cantly shortened the latency and distance of model mice for reaching the platform in the water maze test(P<0.01),and significantly increased the number of crossing the platform(P<0.01);increased the exploration time and number of the Y maze new arm in model mice(P<0.05);inhibited the TUNEL fluorescence expression in the hippocampus of model mice(P<0.01);upregulated the activity of the oxidative stress factor superoxide dismutase(SOD)(P<0.05)and glutathione(GSH)content(P<0.05),and downregulated malondialdehyde(MDA)content(P<0.05);reduced interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF-α)expression levels(P<0.05,P<0.01);decreased the expression of apoptosis signaling pathway proteins Cleaved Caspase-3 and Caspase-3(P<0.05),upregulated the expression of oxidative stress signaling pathway proteins Nrf2 and HO-1(P<0.05),and downregulated the expression of inflammatory stress signaling pathway proteins p-NF-κB and NF-κB(P<0.05).CONCLUSION Qishen Yizhi for-mula can improve the learning and memory ability of subacute aging model mice injected with D-galactose,which may be related to its inhibitory effect on hippocampal oxidative stress and inflammatory stress.

Objective:To investigate the interactive effect of mismatch repair (MMR) protein status and adverse clinicopathological features on prognosis of stage Ⅰ-Ⅲ colon cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 1 650 patients with colon cancer of stage Ⅰ-Ⅲ who were admitted to 7 hospitals in China from January 2016 to December 2017 were collected. There were 963 males and 687 females, aged 62(53,71)years. Patients were classified as 230 cases of MMR deficiency (dMMR) and 1 420 cases of MMR proficiency (pMMR) based on their MMR protein status. Observation indicators: (1) comparison of clinicopathological characteristics between patients of different MMR protein status; (2) analysis of factors affecting the survival outcomes of patients of dMMR; (3) analysis of factors affecting the survival outcomes of patients of pMMR; (4) interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The random forest interpolation method was used for missing values in data interpolation. Univariate analysis was conducted using the COX proportional risk regression model, and multivariate analysis was conducted using the COX stepwise regression with forward method. The coefficient of multiplication interaction effect was obtained using the interaction term coefficient of COX proportional risk regression model. Evaluation of additive interaction effects was conducted using the relative excess risk due to interaction ( RERI). Results:(1) Comparison of clinicopathological characteristics between patients of different MMR protein status. There were significant differences in age, T staging, the number of lymph node harvest, the number of lymph node harvest <12, high grade tumor between patients of dMMR and pMMR ( P<0.05). (2) Analysis of factors affecting the survival outcomes of patients of dMMR. Results of multivariate analysis showed that T staging, N staging, the number of lymph node harvest <12 were independent factors affecting the disease-free survival (DFS) of colon cancer patients of dMMR ( hazard ratio=3.548, 2.589, 6.702, 95% confidence interval as 1.460-8.620, 1.064-6.301, 1.886-23.813, P<0.05). Age and N staging were independent factors affecting the overall survival (OS) of colon cancer patients of dMMR ( hazard ratio=1.073, 10.684, 95% confidence interval as 1.021-1.126, 2.311-49.404, P<0.05). (3) Analysis of factors affecting the survival outcomes of patients of pMMR. Results of multivariate analysis showed that age, T staging, N staging, vascular tumor thrombus were independent factors affecting the DFS of colon cancer patients of pMMR ( hazard ratio=1.018, 2.214, 2.598, 1.549, 95% confidence interval as 1.006-1.030, 1.618-3.030, 1.921-3.513, 1.118-2.147, P<0.05). Age, T staging, N staging, high grade tumor were independent factors affecting the OS of colon cancer patients of pMMR ( hazard ratio=1.036, 2.080, 2.591, 1.615, 95% confidence interval as 1.020-1.052, 1.407-3.075, 1.791-3.748, 1.114-2.341, P<0.05). (4) Interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Results of interaction analysis showed that the multiplication interaction effect between the number of lymph node harvest <12 and MMR protein status was significant on DFS of colon cancer patients ( hazard ratio=3.923, 95% confidence interval as 1.057-14.555, P<0.05). The additive interaction effects between age and MMR protein status, between high grade tumor and MMR protein status were significant on OS of colon cancer patients ( RERI=-0.033, -1.304, 95% confidence interval as -0.049 to -0.018, -2.462 to -0.146). Conclusions:There is an interaction between the MMR protein status and the adverse clinicopathological features (the number of lymph node harvest <12, high grade tumor) on prognosis of colon cancer patients of stage Ⅰ-Ⅲ. In patients of dMMR, the number of lymph node harvest <12 has a stronger predictive effect on poor prognosis. In patients of pMMR, the high grade tumor has a stronger predictive effect on poor prognosis.

Objective To investigate the status quo of pain catastrophizing(PC)in the patients with di-abetic peripheral neuropathic pain(DPNP),and to analyze the influencing factors to provide reference for for-mulating clinical preventive intervention strategies.Methods A total of 206 patients with DPNP admitted and treated in the People's Hospital of Guangxi Zhuang Autonomous Region were selected as the research sub-jects by convenience sampling method.The general data questionnaire,Numerical Rating Scale(NRS),Pain Catastrophizing scale(PCS),Perceived Social Support Scale(PSSS)and diabetes distress scale(DDS)were used to conduct the investigation.Results The incidence rate of PC in 206 cases of DPNP patients was 44.66％(92/206),and the total score of PCS was(30.10±5.16)points.The results of multiple linear regres-sion analysis showed that the gender,duration of diabetes(≥10 years),multiple drug use,number of compli-cations(＞5),NRS score,PSSS score and scores of DDS dimensions were the main influencing factors of PC(all P＜0.05),which could explain 92.3％of the total variation of PC.Conclusion The PC incidence rate in the patients with DPNP is high.Clinical healthcare workers should pay attention to the evaluation of PC in these patients,and formulate the scientific and effective targeted intervention measures according to the main influen-cing factors to help the patients to reduce the pain burden in order to reduce the level of PC.

Objective:To evaluate the characteristics of vaginal flora between normal and abnormal pregnant women throughout pregnancy.Methods:Vaginal swab specimens were collected from pregnant women in the first (<14 gestation weeks, GW), second (14~28 GW) and third trimester (>28 GW) in Anqing, Anhui Province from February 2018 to February 2020. Pregnant women were divided into normal and abnormal groups according to all clinical diagnosis. The sequences of 16S rRNA gene (V3-V4) from vaginal swabs were analyzed using QIIME2 platform. The differences in the dominance of Lactobacillus, community state type (CST) transition, Alpha diversity and Beta diversity were analyzed. Diversity data after log transition were used in the analysis of linear mixed model. Results:A total of 34 pregnant women (10 normal and 24 abnormal) with 102 samples were included for analysis. The composition of vaginal flora between two groups: the relative abundance of Lactobacillus was the highest at the genus level and Lactobacillus crispatus and Lactobacillus iners was the top two species with high relative abundance. The dominance of Lactobacillus, Alpha diversity and transition of CST were also similar. Both groups had a gradually decreased trend of Alpha diversity with GW, and the Chao1, Observed species and Faith′s PD indexes′ were different in different GW ( P<0.05). All Beta diversity metrics in normal group had descending trend, with lower value of the index of first distance which implied a higher microbiota stability, while Bray-Curtis, Weighted UniFrac distance had ascending trend in abnormal group, indicating lower stability. Jaccard distance′s first distance was statistically differed among GW and Unweighted UniFrac distance′s differed between normal and abnormal groups. Conclusions:The first distance of Unweighte UniFrac distance in abnormal pregnant women is higher than that of normal pregnant women and the vaginal flora in abnormal group has lower stability.

Objective To investigate the effects of tetramethylpyrazine(TMP)on acute kidney injury(AKI)induced by cisplatin(Cis)and its related mechanisms.Methods Sixty SPF male C57BL/6J mice were randomly divided into control group(Ctl),TMP group,Cis-induced AKI model group(Cis group)and TMP treated AKI model group(Cis+TMP group).The contents of BUN and Scr were detected,and the apoptosis of renal tissue was detected by TUNEL staining.The structural changes of mitochondria in renal cortex of each group were observed by transmission electron microscope,and the expression of ROS in renal tissue was detected by DHE staining.The expression of PGC-1α and the phosphorylation of Drp1(p-Drp1Ser637/Drp1)in renal tissue were detected by Western blotting assay.Proximal renal tubule epithelial cell line HK-2 was cultured in vitro and divided into Ctl group,TMP group,Cis group and Cis+TMP group.Proliferation activity of cells in each group was detected by CCK-8 assay,apoptosis was detected by Annexin V-FITC/PI method,ROS expression and mitochondrial membrane potential were detected by DCFH-DA fluorescence probe and JC-1 staining.The expression of PGC-1α and p-Drp1Ser637/Drp1 were detected by Western blotting assay.Results Compared with Ctl group,the contents of BUN and Scr in Cis group and Cis+TMP group were significantly increased(P<0.05),cell apoptosis of renal tubular and the damage of renal cortical mitochondrial structure were significantly increased(P<0.05).However,the expression of PGC-1α protein and p-Drp1Ser637/Drp1 ratio in renal tissue were significantly decreased(P<0.05),and no significant changes were observed in TMP group(P>0.05).Compared with Cis group,the change trend of above detection indexes in Cis+TMP group was significantly decreased(P<0.05).Compared with HK-2 cells in Ctl group,the proliferation activity,mitochondrial membrane potential,PGC-1α protein expression and p-Drp1Ser637/Drp1 ratio of Cis group and Cis+TMP group were significantly decreased(P<0.05),while the apoptosis ratio of Cis and Cis+ TMP group was significantly decreased(P<0.05).The ROS expression and Drp1 protein expression in mitochondria were significantly increased(P<0.05),but no significant changes were observed in TMP group(P>0.05).Compared with Cis group,the change trend of above detection indexes in Cis+TMP group was significantly decreased(P<0.05).Conclusions TMP can ameliorate CIS-induced apoptosis and mitochondrial damage in renal tubular epithelial cells in vitro and in vivo,which may be related to PGC-1α pathway.

Objective:To investigate the therapeutic efficiency of multiple forms of microdissected perforator flaps in the reconstruction of limb defect.Methods:The data of patients treated in the Department of Hand Surgery of Xuzhou Renci Hospital from June 2020 to June 2022 for repairing wound of hands, feet, limbs and popliteal fosses with microdissected perforator flap were retrospectively analyzed. CT angiography and color Doppler ultrasound were performed in all patients before surgery to locate perforator vessels, design the donor flap, and cut the flap retrograde according to the design line. After confirming good blood supple, the flap was harvested under the microscope, and fat particles were cut around the vascular pedicle by "insect eating" technique. After that, the flap was transferred to the recipient area, completed the vascular anastomosis, and placed the drainage tube. The donor site was closed directly. The blood circulation of the flap was observed closely after operation. Regular follow-up visits were conducted in outpatient clinics, door-to-door visits and WeChat to record sensory recovery and complications. Postoperative wound healing, flap shape, flap sensation, flap temperature and scar in donor area were evaluated using the efficacy satisfaction scale, which was divided into four grades: excellent (2 points), good (1 point), fair (0 point) and poor (-1 point). The total score of 5-10 was satisfactory, 0-4 was average, and -1--5 was unsatisfactory.Results:A total of 16 patients were included, including 8 males and 8 females. The age ranged from 26 to 65 years, with an average age of 45.9 years. There were 2 cases of upper limb soft tissue defect, 2 cases of hand degloving injury, 11 cases of hand skin soft tissue defect, and 1 case of popliteal soft tissue defect. The wound size ranged from 3.0 cm×4.5 cm to 13.0 cm×30.0 cm. Free medial sural artery perforator flap was used in 4 cases, interosseous dorsal artery perforator flap in 1 case, anterolateral femoral perforator flap in 6 cases, thoracic umbilical flap in 1 case, anterolateral femoral perforator flap combined with superficial circumflex iliac artery perforator flap in 1 case, and superficial circumflex iliac artery perforator flap in 3 cases. The size of the skin flap was 3.5 cm×5.0 cm - 9.5 cm×30.0 cm. After operation, 15 cases of flaps successfully survived, 1 case of venous crisis occurred 1 day after operation, and successfully survived after exploration and re-anastomosis of blood vessels. The wounds in the donor area healed in one stage without wound infection, delayed healing and other complications. All patients were follow-up for 7 to 18 months (mean 12.1 months), the skin flap survived well, no pigmentation, bloated appearance, and no patients requested secondary thinning treatment. The skin flap sensation recovered light touch in 1 case, protective sense in 9 cases and deep touch only in 6 cases. No significant thickening of skin flap was observed in the later period of follow-up, and skin flap wear was observed in 1 patient. There were only linear scars left in the donor area, and there were no obvious abnormalities in appearance and function. Efficacy satisfaction evaluation showed that all patients were satisfied with the therapeutic effect, scoring 6-9 points, with an average of 7.8 points.Conclusion:Microdissected perforator flap technique provides a one-stage surgical solution for aesthetic and functional areas with high requirements, and satisfactory clinical efficiencies can be obtained.

Objective:To investigate the therapeutic efficiency of multiple forms of microdissected perforator flaps in the reconstruction of limb defect.Methods:The data of patients treated in the Department of Hand Surgery of Xuzhou Renci Hospital from June 2020 to June 2022 for repairing wound of hands, feet, limbs and popliteal fosses with microdissected perforator flap were retrospectively analyzed. CT angiography and color Doppler ultrasound were performed in all patients before surgery to locate perforator vessels, design the donor flap, and cut the flap retrograde according to the design line. After confirming good blood supple, the flap was harvested under the microscope, and fat particles were cut around the vascular pedicle by "insect eating" technique. After that, the flap was transferred to the recipient area, completed the vascular anastomosis, and placed the drainage tube. The donor site was closed directly. The blood circulation of the flap was observed closely after operation. Regular follow-up visits were conducted in outpatient clinics, door-to-door visits and WeChat to record sensory recovery and complications. Postoperative wound healing, flap shape, flap sensation, flap temperature and scar in donor area were evaluated using the efficacy satisfaction scale, which was divided into four grades: excellent (2 points), good (1 point), fair (0 point) and poor (-1 point). The total score of 5-10 was satisfactory, 0-4 was average, and -1--5 was unsatisfactory.Results:A total of 16 patients were included, including 8 males and 8 females. The age ranged from 26 to 65 years, with an average age of 45.9 years. There were 2 cases of upper limb soft tissue defect, 2 cases of hand degloving injury, 11 cases of hand skin soft tissue defect, and 1 case of popliteal soft tissue defect. The wound size ranged from 3.0 cm×4.5 cm to 13.0 cm×30.0 cm. Free medial sural artery perforator flap was used in 4 cases, interosseous dorsal artery perforator flap in 1 case, anterolateral femoral perforator flap in 6 cases, thoracic umbilical flap in 1 case, anterolateral femoral perforator flap combined with superficial circumflex iliac artery perforator flap in 1 case, and superficial circumflex iliac artery perforator flap in 3 cases. The size of the skin flap was 3.5 cm×5.0 cm - 9.5 cm×30.0 cm. After operation, 15 cases of flaps successfully survived, 1 case of venous crisis occurred 1 day after operation, and successfully survived after exploration and re-anastomosis of blood vessels. The wounds in the donor area healed in one stage without wound infection, delayed healing and other complications. All patients were follow-up for 7 to 18 months (mean 12.1 months), the skin flap survived well, no pigmentation, bloated appearance, and no patients requested secondary thinning treatment. The skin flap sensation recovered light touch in 1 case, protective sense in 9 cases and deep touch only in 6 cases. No significant thickening of skin flap was observed in the later period of follow-up, and skin flap wear was observed in 1 patient. There were only linear scars left in the donor area, and there were no obvious abnormalities in appearance and function. Efficacy satisfaction evaluation showed that all patients were satisfied with the therapeutic effect, scoring 6-9 points, with an average of 7.8 points.Conclusion:Microdissected perforator flap technique provides a one-stage surgical solution for aesthetic and functional areas with high requirements, and satisfactory clinical efficiencies can be obtained.

Objective:To analyze the online learning behaviors and learning results of students under different teaching modes with different learning motivations, and to provide a reference for optimizing the development of online teaching.Methods:Samples of students taking the physiology small private online course (SPOC) in 2019 and 2020, and students taking the massive open online courses (MOOC) in the same year were selected in the study to discuss the differences in learning behaviors of students under different teaching modes and learning effects on students with different learning motivations. The online learning behaviors of students were analyzed based on their learning engagement, interactive behavior and learning motivation. SPSS 25.0 was used to analyze the data with statistical methods such as function calculation, frequency statistics, regression analysis, rank sum test, correlation test and chi-square test.Results:The learning participation level ( z=14.36, P<0.001), and the human-machine and interpersonal learning interaction level ( z=-11.70, -16.18, both P<0.001) of SPOC learners in 2020 were higher than those in 2019. The overall interactive level was moderately correlated with performances of students ( r=0.42, 0.52, both P<0.001), and the correlation between interpersonal behavior and grades was much more stronger ( r=0.60, 0.55, both P<0.001). The performance composition of SPOC and MOOC learners in 2019 was completely different ( χ2=857.45, P<0.001). The learning effect of externally motivated students was significantly better than that of internally motivated students ( z=-28.42, P<0.001). Conclusion:The teaching mode adopted by the school can affect the learning effect by influencing the students' online learning engagement and interactive behavior. Besides, students' own learning motivation also plays a key role in their academic performance.