OBJECTIVE To study the mechanism of Wenshen Tongdu Recipe in promoting the recovery of motor function in rats with spinal cord injury.METHODS A total of 144 SD rats were randomly divided into sham operation group,model group,predni-sone group,low-dose Wenshen Tongdu Recipe group,medium-dose Wenshen Tongdu Recipe group,high-dose Wenshen Tongdu Recipe group,3BDO group,and 3BDO+Wenshen Tongdu Recipe group(medium dose).The rat spinal cord injury model was estab-lished by modified Allen's method.Intervention began 1 day after modeling,and the drug was administered continuously for 14 days.During the drug administration period,the motor function of the rats in each group was evaluated by Basso-Beattie-Bresnahan(BBB)score and inclined plane test.On the 3rd,7th and 14th days after modeling,the pathological changes of the spinal cord tissues of the rats in each group were observed by HE staining;the expression levels of inflammatory factors IL-1β,IL-6,IL-4 and IL-10 in the serum of the rats in each group were detected by ELISA;the expression of Beclin 1,LC3B and p62 proteins was detected by immu-nohistochemistry;the expression of CD16 and CD206 proteins was detected by immunofluorescence staining;the expression of autoph-agy-related molecules(Beclin 1,LC3B)and Akt/mTOR pathway-related proteins in the spinal cord tissues was detected by Western blot.RESULTS Compared with the model group,the BBB score and angle of the inclined board test of rats in the medium-dose Wenshen Tongdu Recipe group were increased,and the disordered arrangement of spinal cord tissue,spinal cord vacuoles and inflam-matory infiltration were significantly improved,especially on the 14th day.Compared with the sham operation group,the expression levels of serum IL-1β and IL-6 in the model group increased(P<0.01),and the expression levels of IL-4 and IL-10 decreased(P<0.05,P<0.01);compared with the model group,the levels of IL-1β and IL-6 in the Wenshen Tongdu Recipe group were signifi-cantly decreased(P<0.01),and the levels of IL-4 and IL-10 were significantly increased(P<0.05,P<0.01);compared with the Wenshen Tongdu Recipe group,the serum IL-1β and IL-6 concentrations of mice in the 3BDO group increased(P<0.01),and the level of IL-10 decreased(P<0.05).Compared with the sham operation group,the M1/M2 ratio,P62 protein expression,p-Akt/Akt and p-mTOR/mTOR ratios in the spinal cord tissue of the rats in the model group were significantly increased(P<0.05,P<0.01),and the relative protein expression of Beclin1 was decreased(P<0.01);compared with the model group,the M1/M2 ratio,p-Akt/Akt and p-p70S6K/p70S6K ratios in the Wenshen Tongdu Recipe group were significantly decreased(P<0.01).Compared with the model group,the Beclin1 protein level in the 3BDO group was decreased,and the p-Akt/Akt and p-mTOR/mTOR ratios were increased(P<0.05,P<0.01).Compared with the Wenshen Tongdu Recipe group,the M1/M2 ratio in the 3BDO group and the 3BDO+Wen-shen Tongdu Recipe group increased(P<0.01),the positive rates of Beclin1 and LC3B proteins in the 3BDO group decreased signifi-cantly(P<0.01),and the p-p70S6K/p70S6K ratio in the 3BDO+Wenshen Tongdu Recipe group increased significantly(P<0.01).CONCLUSION Wenshen Tongdu Recipe may regulate microglial polarization through Akt/mTOR signaling pathway to acti-vate autophagy,promote the secretion of anti-inflammatory factors,reduce the release of pro-inflammatory factors,alleviate neuroin-flammatory response,and thus promote spinal cord injury repair.

OBJECTIVE To study the mechanism of Wenshen Tongdu Recipe in promoting the recovery of motor function in rats with spinal cord injury.METHODS A total of 144 SD rats were randomly divided into sham operation group,model group,predni-sone group,low-dose Wenshen Tongdu Recipe group,medium-dose Wenshen Tongdu Recipe group,high-dose Wenshen Tongdu Recipe group,3BDO group,and 3BDO+Wenshen Tongdu Recipe group(medium dose).The rat spinal cord injury model was estab-lished by modified Allen's method.Intervention began 1 day after modeling,and the drug was administered continuously for 14 days.During the drug administration period,the motor function of the rats in each group was evaluated by Basso-Beattie-Bresnahan(BBB)score and inclined plane test.On the 3rd,7th and 14th days after modeling,the pathological changes of the spinal cord tissues of the rats in each group were observed by HE staining;the expression levels of inflammatory factors IL-1β,IL-6,IL-4 and IL-10 in the serum of the rats in each group were detected by ELISA;the expression of Beclin 1,LC3B and p62 proteins was detected by immu-nohistochemistry;the expression of CD16 and CD206 proteins was detected by immunofluorescence staining;the expression of autoph-agy-related molecules(Beclin 1,LC3B)and Akt/mTOR pathway-related proteins in the spinal cord tissues was detected by Western blot.RESULTS Compared with the model group,the BBB score and angle of the inclined board test of rats in the medium-dose Wenshen Tongdu Recipe group were increased,and the disordered arrangement of spinal cord tissue,spinal cord vacuoles and inflam-matory infiltration were significantly improved,especially on the 14th day.Compared with the sham operation group,the expression levels of serum IL-1β and IL-6 in the model group increased(P<0.01),and the expression levels of IL-4 and IL-10 decreased(P<0.05,P<0.01);compared with the model group,the levels of IL-1β and IL-6 in the Wenshen Tongdu Recipe group were signifi-cantly decreased(P<0.01),and the levels of IL-4 and IL-10 were significantly increased(P<0.05,P<0.01);compared with the Wenshen Tongdu Recipe group,the serum IL-1β and IL-6 concentrations of mice in the 3BDO group increased(P<0.01),and the level of IL-10 decreased(P<0.05).Compared with the sham operation group,the M1/M2 ratio,P62 protein expression,p-Akt/Akt and p-mTOR/mTOR ratios in the spinal cord tissue of the rats in the model group were significantly increased(P<0.05,P<0.01),and the relative protein expression of Beclin1 was decreased(P<0.01);compared with the model group,the M1/M2 ratio,p-Akt/Akt and p-p70S6K/p70S6K ratios in the Wenshen Tongdu Recipe group were significantly decreased(P<0.01).Compared with the model group,the Beclin1 protein level in the 3BDO group was decreased,and the p-Akt/Akt and p-mTOR/mTOR ratios were increased(P<0.05,P<0.01).Compared with the Wenshen Tongdu Recipe group,the M1/M2 ratio in the 3BDO group and the 3BDO+Wen-shen Tongdu Recipe group increased(P<0.01),the positive rates of Beclin1 and LC3B proteins in the 3BDO group decreased signifi-cantly(P<0.01),and the p-p70S6K/p70S6K ratio in the 3BDO+Wenshen Tongdu Recipe group increased significantly(P<0.01).CONCLUSION Wenshen Tongdu Recipe may regulate microglial polarization through Akt/mTOR signaling pathway to acti-vate autophagy,promote the secretion of anti-inflammatory factors,reduce the release of pro-inflammatory factors,alleviate neuroin-flammatory response,and thus promote spinal cord injury repair.

OBJECTIVE To study the structural characteristics of purified polysaccharide isolated from Polygonatum cyrtonema Hua and explore its anti-fatigue and intestinal bacteria improvement effects,so as to provide theoretical basis for the development of products using this polysaccharide as functional ingredients.METHODS PMP pre-column derivatization-high performance liquid chromatography,trimethylsilyl ether derivatization-gas chromatography and other methods were used to chemically characterize the pu-rified polysaccharide;the mouse weight-loading swimming test and biochemical indicators were used to evaluate its anti-fatigue effect;16S rRNA intestinal flora sequencing was adopted to study the effect of the purified product in improving intestinal bacteria.RESULTS The obtained polysaccharide was a fructan product and contained small amounts of glucose(Glc),galactose(Gal),arabinose(Ara),mannose(Man),rhamnose(Rha)and galacturonic acid(GalA),which had a relative molecular mass in the range of 0.4-285 kDa,and the molecular weight of the main component was around 3 kDa.Compared with the model group,the high-dose polysaccha-ride group could significantly prolong the exhaustive swimming time of mice(P＜0.01),reduce the levels of BUN and UA in serum and the LA(P＜0.01)and MDA levels(P＜0.05)in muscle tissue,and enhance SOD(P＜0.01),GSH-Px(P＜0.001)and CAT(P＜0.05)levels in liver tissue.Compared with the blank control group,the low-dose polysaccharide group could increase the relative a-bundance of probiotics Eggerthelaceae(g_unclassified_f_Eggerthelaceae)and Weissella(g_Weissella)in the fecal microorganisms of mice(P＜0.05).CONCLUSION The chemical composition of purified polysaccharide isolated from Polygonatum cyrtonema Hua is determined,and it is proven that the polysaccharide has a good anti-fatigue effect and can promote the growth of probiotics in the intes-tine,which provides a basis for product development using this polysaccharide as a functional ingredient.

Objective:To explore the effects of pharmaceutical services through enterprise WeChat for outpatients with cancer pain.Methods:A retrospective study was conducted on patients who were first diagnosed and/or previously treated for cancer pain and visited Outpatient Department of the First Affiliated Hospital of Soochow University from January 1, 2020 to January 30, 2023. The outpatients were divided into conventional pharmaceutical services management group (conventional group) and pharmaceutical services management group through enterprise WeChat (enterprise WeChat group) based on whether they received pharmaceutical services for cancer pain through enterprise WeChat. Prescription appropriateness, changes in cancer pain and life quality evaluation indicators after treatment in patients between the 2 groups were recorded and compared.Results:A total of 174 patients were included, with 87 patients in each group. There were no significant differences in age and gender between the 2 groups (all P>0.05). Before the pharmaceutical services management for cancer pain through enterprise WeChat, the differences in the proportion of patients with pain free, mild, moderate, and severe pain after pain relief treatments between the 2 groups were not significant (all P>0.05). In the conventional group, 13 out of 87 patients (14.9%) had inappropriate prescriptions, and 2 out of 87 (2.3%) in the enterprise WeChat group, with statistically significant difference (χ 2=8.828, P<0.05). After management with enterprise WeChat, the patients of pain-free increased from 19 to 55 (63.2%) in the enterprise WeChat group, and from 18 to 41 (47.1%) in the conventional group. The difference in the proportion of pain-free patients between the 2 groups was statistically significant (χ 2=4.555, P=0.033). The differences in the scores of various indicators of life quality between the 2 groups before management with enterprise WeChat were not significant (all P>0.05). After the management with enterprise WeChat, life quality scores in all the 8 dimensions were significantly higher than those in the conventional group (all P<0.05). Conclusion:The utilization of enterprise WeChat could improve the treatment effect of cancer pain, enhance their life quality, help improve the quality of pharmaceutical services, and have certain promotion and utilization value.

Objective:To explore the effects of pharmaceutical services through enterprise WeChat for outpatients with cancer pain.Methods:A retrospective study was conducted on patients who were first diagnosed and/or previously treated for cancer pain and visited Outpatient Department of the First Affiliated Hospital of Soochow University from January 1, 2020 to January 30, 2023. The outpatients were divided into conventional pharmaceutical services management group (conventional group) and pharmaceutical services management group through enterprise WeChat (enterprise WeChat group) based on whether they received pharmaceutical services for cancer pain through enterprise WeChat. Prescription appropriateness, changes in cancer pain and life quality evaluation indicators after treatment in patients between the 2 groups were recorded and compared.Results:A total of 174 patients were included, with 87 patients in each group. There were no significant differences in age and gender between the 2 groups (all P>0.05). Before the pharmaceutical services management for cancer pain through enterprise WeChat, the differences in the proportion of patients with pain free, mild, moderate, and severe pain after pain relief treatments between the 2 groups were not significant (all P>0.05). In the conventional group, 13 out of 87 patients (14.9%) had inappropriate prescriptions, and 2 out of 87 (2.3%) in the enterprise WeChat group, with statistically significant difference (χ 2=8.828, P<0.05). After management with enterprise WeChat, the patients of pain-free increased from 19 to 55 (63.2%) in the enterprise WeChat group, and from 18 to 41 (47.1%) in the conventional group. The difference in the proportion of pain-free patients between the 2 groups was statistically significant (χ 2=4.555, P=0.033). The differences in the scores of various indicators of life quality between the 2 groups before management with enterprise WeChat were not significant (all P>0.05). After the management with enterprise WeChat, life quality scores in all the 8 dimensions were significantly higher than those in the conventional group (all P<0.05). Conclusion:The utilization of enterprise WeChat could improve the treatment effect of cancer pain, enhance their life quality, help improve the quality of pharmaceutical services, and have certain promotion and utilization value.

Objective To prepare and identify rabbit anti-cyclin dependent kinase 6 (CDK6) antibody. Methods The recombinant pET21a (+)/CDK6 was successfully constructed, then the recombinant plasmid was transformed into E.coli BL21 (DE3) competent cells and was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) for protein expression, which was detected by SDS-PAGE and Western blot analysis. The expressed protein was purified by nickel-chelating nitrilotriacetic acid (Ni-NTA) agarose and then analyzed by SDS-PAGE. Japanese white rabbits were immunized with purified CDK6 protein for many times every two weeks. The blood was collected at 0, 2, 4 and 6 weeks after immunization, and serum was separated from blood. The titer was detected by indirect ELISA. Western blot analysis, immunofluorescence assay and immunohistochemistry were employed to determine the specificity. Results High purity CDK6 protein and high specificity of rabbit anti-CDK6 antibody were successfully prepared. The titer of CDK6 rabbit serum antibody reached 1:30 000 after immunization, which could specifically recognize the CDK6 protein expressed in cervical cancer cell line and cervical cancer tissues. Conclusion The high titer and specificity of rabbit anti-CDK6 antibody is successfully prepared.
Animals ; Female ; Humans ; Rabbits ; Antibodies ; Antibody Specificity ; Blotting, Western ; Cyclin-Dependent Kinase 6 ; Enzyme-Linked Immunosorbent Assay ; Uterine Cervical Neoplasms

Objective:To investigate the status of frailty and activities of daily living (ADL) in maintenance hemodialysis (MHD) patients, and to explore the effect of frailty phenotype on ADL.Methods:The patients who underwent MHD in Kidney Disease Center of the First Affiliated Hospital from March 2019 to March 2020 were enrolled in this study. The demographic and laboratorial data were collected by cross-sectional survey method. Fried frailty phenotype scale and ADL scale were used to evaluate the frailty and ADL, respectively. The differences of basic data and different frailty phenotypes between the normal function group and the function decline group were compared in terms of ADL, physical self-maintenance ability and instrumental ADL ability. Pearson correlation analysis was used to analyze the correlation between frailty and ADL, and binary logistic regression analysis was used to analyze the influencing factors of ADL.Results:A total of 676 MHD patients were included in this study, including 434 males (64.2%) and 242 females (35.8%). The age was (59.2±19.4) years old, and the median dialysis age was 59.0 (25.3, 110.0) months. There were 159 frailty patients (23.5%), 230 pre-frailty patients (34.0%), and 287 non-frailty patients (42.5%). The ADL was decreased in 163 patients (24.1%), including 131 patients (19.4%) with decreased physical self-maintenance ability and 161 patients (23.8%) with decreased instrumental ADL ability. Pearson correlation analysis showed that the frailty score was positively correlated with total ADL score ( r=0.728, P<0.001), physical self-maintenance ability score ( r=0.669, P<0.001) and instrumental ADL ability score ( r=0.729, P<0.001). Binary logistic regression analysis results showed that older age and lower physical activity, fatigue, slowed steps and lower grip strength in the frailty phenotypes were the independent influencing factors of ADL, physical self-maintenance ability and instrumental ADL ability (all P<0.05). Conclusions:The prevalence of frailty is 23.5% in MHD patients, and 24.1% of MHD patients have decreased ADL. Elder age and lower physical activity, fatigue, reduced step counts, and lower grip strength in frailty phenotypes are the independent influencing factors for poor ADL, poor physical self-maintenance ability and poor instrumental ADL ability.

Objective: To investigate the influencing factors of smoking tendency of adolescents and to analyze the influence of e-cigarette on smoking tendency of teenagers and the possible interaction, to provide evidence for tobacco control measures. Methods: Stratified multistage cluster probability sampling method was used to select 6 178 students from junior middle school, high school and vocational high school students in Shanghai for questionnaire survey from September to October 2019. Students with smoking tendency were taken as case group, adolescents with same sex and similar age(within 1 year) were mathed in 1∶3 ratio. A total of 631 adolescents with smoking tendency and 1 870 non-smoking tendency were included in the study. Conditional Logistic regression was used to analyze the influencing factors and possible interactions. Results: After adjusting gender, age and other covariates, conditional Logistic analysis results showed that the smoking tendency of middle school students was associated with factors such as their friends smoking, trying cigarettes and trying e-cigarettes. The OR value and 95%CI were 3.26(2.47-4.29), 5.90(3.76-9.24), and 3.28(2.11-5.10), respectively. The interaction analysis results showed that the OR value and 95%CI of the multiplying interaction between friends smoking and trying e-cigarettes, trying smoking and trying e-cigarettes were 8.62(4.90-15.17) and 12.01(6.02-23.95), respectively. There was no additive interaction. Conclusion Tobacco control interventions, such as e-cigarette harm publicity and peer education, can help to change teenagers smoking tendency and further reduce their tobacco use rate.

Objective To evaluate the position of the fetal conus medullaris during pregnancy and its value in detecting tethered cord syndrome(TCS). Methods Nine hundred and seventy-four normal fetuses and 46 fetuses with TCS between 15 and 41 weeks gestation were involved in the study.Parameters D 1 (the distance between the end of the conus medullaris and the caudal edge of last vertebral body ossification center) and D2 (the distance from the end of the conus medullaris to the caudal skin namely the intersection point of the extending line of D1 and the skin) were measured in the caudal midsagittal plane of the spine. Sixty normal fetuses were chosed randomly for interobserver variability.Correlation analysis between these two parameters and gestational age(GA) were conducted and the normal reference value of these parameters were calculated in normal group. The ratios of growth parameters ( Biparietal diameter, Head circumference,Abdominal circumference,Femur length) to D1 and D2 were calculated separately to observe the difference of the ratios between two groups. All the parameters and ratios of normal fetuses were compared with that of TCS cases.Results There was no significant difference in D1 and D2 between two observers.A significant linear correlation between the parameters and GA was found in normal group,linear regression equations were D1=0.251 GA -2.265 cm (R2=0.926,P <0.01) and D2=0.267 GA -1.812 cm(R2=0.928,P <0.01),respectively.D1 and D2 were much lower in normal group than in abnormal group (all P <0.01). The ratios of the growth parameters to D1 and D2 were relatively stable and had statistically differences between two groups in different gestational age. Conclusions The methods that determination of D1 and D2 are simple and feasible,and could help to the prenatal diagnosis of TCS.

OBJECTIVE: To complete standardized management and follow-up for the cancer pain and chronic pain patients in outpatient department. METHODS: Information management platform was designed and established for cancer pain and chronic pain patients in outpatient department, including information entry, information inquiry, prompt for drug dispensing in advance, file unified management and regular scanning and removing excess medical records. The procedure of prescribing narcotic drugs for first visiting, subsequent visiting and follow-up patients was introduced. File management and prescription rationality before and after the establishment of information platform were compared. The adoption of pharmacist' s recommendation after the establishment of information platform were counted. RESULTS: Compared with before the establishment of information platform, the ratio of problematic profiles decreased from 42. 96％ to 7. 06％ (P＜0. 05); the ratio of irrational prescription decreased from 4. 39％ to 1. 19％ (P＜0. 05). Pharmacists provided a total of 1 542 recommendations for the patients with poor pain control and ADR. The adoption rate of the pharmaceutical recommendations was 63. 62％. CONCLUSIONS: The cancer pain and chronic pain information management platform can manage the outpatients' profile effectively and improve the rationality of the prescription. Meanwhile, the pharmacists have played an important role in the pain treatment for outpatients.