BACKGROUND:Macrophage migration inhibitory factor(MIF)is a pleiotropic cytokine,which is secreted in different types of stem cells and can regulate the proliferation,differentiation and migration of various types of stem cells.Our previous research has confirmed that human embryonic stem cells secrete MIF and that its concentration in the culture medium is relatively stable.However,whether MIF is involved in the survival,proliferation and differentiation of human embryonic stem cells remains unclear. OBJECTIVE:To investigate the effects of MIF on survival,proliferation,and differentiation of human embryonic stem cells. METHODS:(1)Human embryonic stem cells H9 were cultured.The growth curve of cells was detected and plotted by CCK-8 assay.Enzyme-linked immunosorbent assay was used to determine the level of MIF in the medium.(2)To determine the effects of exogenous MIF on the survival and proliferation of human embryonic stem cells,different groups were established:the control group,which was cultured in stem cell medium without any modifications;the exogenous MIF group,which was treated with different concentrations(30,100,300 ng/mL)of MIF in the stem cell medium;the MIF inhibitor ISO-1 group,which was treated with different concentrations(2,7,21 μmol/L)of ISO-1 in the stem cell medium;and the MIF+ISO-1 group,which was treated with different concentrations of ISO-1 along with 100 ng/mL of MIF.Cell viability was assessed using the CCK-8 assay.(3)To further elucidate the effect of MIF gene on survival and proliferation of human embryonic stem cell,the MIF knockout H9 cell line was constructed by CRISPR-Cas 9 technology to observe the lineage establishment.(4)To determine the effect of high concentrations of MIF on human embryonic stem cell differentiation,100 ng/mL MIF and 100 ng/mL of CXCR4 neutralizing antibody were separately added to the normal stem cell culture medium.The expression levels of self-renewal factors(KLF4,c-MYC,NANOG,OCT4,and SOX2)and differentiation transcription factors(FOXA2,OTX2)were measured using real-time quantitative polymerase chain reaction,immunofluorescence staining,and western blot analysis. RESULTS AND CONCLUSION:(1)The logarithmic growth phase of H9 cells was between 3-6 days.Under normal growth conditions,human embryonic stem cells secreted MIF at a concentration of approximately 20 ng/mL,independent of cell quantity.(2)Compared to the control group,the addition of different concentrations of MIF had no effect on the proliferation of human embryonic stem cells(P>0.05).ISO-1 significantly inhibited the proliferation of human embryonic stem cells,with a stronger inhibition observed at higher concentrations of ISO-1(P<0.05).The addition of MIF in the presence of ISO-1 reduced the inhibitory effect of ISO-1(P<0.05).(3)Real-time quantitative polymerase chain reaction showed that knocking out 50%of the MIF gene resulted in a significant decrease in the growth vitality of human embryonic stem cells and failure to establish cell lines.(4)Adding 100 ng/mL exogenous MIF to the culture medium resulted in a decrease in the mRNA,protein,and fluorescence expression levels of the self-renewal transcription factor KLF4,while the mRNA,protein,and fluorescence expression levels of the differentiation factor FOXA2 increased.(5)When 100 ng/mL CXCR4 neutralizing antibody was added to the culture medium,the mRNA and protein expression levels of KLF4 increased,while the mRNA and protein expression levels of FOXA2 decreased,contrary to the expression trend observed in the MIF group.In conclusion,the endogenous secretion of MIF by human embryonic stem cells is essential for their survival.The addition of MIF to the culture medium does not promote the proliferation of human embryonic stem cells.However,it can lead to a decrease in the expression of the self-renewal factor KLF4 and an increase in the expression of the transcription factor FOXA2.This provides a clue for further investigation into the effects and mechanisms of MIF on the differentiation of human embryonic stem cells.The MIF-CXCR4 axis plays a regulatory role in this process.

ObjectiveTo explore the possible mechanism of Modified Gegen Qinlian Decoction （加味葛根芩连汤， MGQD） in the treatment of ulcerative colitis （UC） based on intestinal mucus barrier. MethodsThirty C57BL/6 mice were randomly divided into a control group， a model group and a MGQD group with 10 mice in each. Dextran Sulfate Sodium Salt （DSS） was used to construct the UC model in all groups except for the control group. Meanwhile， mice in the MGQD group were given 20 g/kg of MGQD decoction by gavage according to their body weight， while those in the control group and model group were given 0.2 ml/20 g of pure water by gavage， once a day for 7 consecutive days. On the day following the last gavage， the body weight， disease activity index （DAI） score， spleen weight， and colon length were compared. The pathological changes of the intestinal mucosal tissues were observed by HE staining； the protein expression levels of mucin 2 （MUC2） and leucine-rich repeat G protein-coupled receptor 5 （Lgr5） in the intestinal mucosal tissues were detected by immunofluorescence； the cuprocytes in the intestinal mucosal tissues were detected by AB/PAS staining； and the expression level of Ki67 in the intestinal mucosal tissues was detected by immunohistochemistry. ResultsHE staining showed that the colon mucosal tissue of the mice in the control group was intact. In the model group， the colon mucosal epithelial structure was severely damaged， with a large amount of inflammatory cell infiltration in the mucosal propria. In the MGQD group， the mucosal tissue structure was partially lost， with a small amount of inflammatory cell infiltration.The body weight and colon length of mice in the model group decreased significantly compared to those in the control group， while DAI scores and spleen weight increased， and the levels of MUC2， Ki67， Lgr5 proteins， and the number of goblet cells were significantly reduced （P＜0.01）. Compared to the model group， the MGQD group had increased body weight of mice， colon length， and decreased DAI scores and spleen weight； the levels of MUC2， Ki67， Lgr5 proteins， and the number of goblet cells were increased （P＜0.05 or P＜0.01）. ConclusionMGQD has a favorable ameliorative effect on UC-related symptoms and pathological tissue damage， and its mechanism of action may be related to the restoration of the prolife-ration and differentiation of intestinal stem cells into goblet cells， thereby promoting the repair of the intestinal mucus barrier.

Objective To investigate the effects of amlodipine benzenesulfonate tablets administered at different time points on blood pressure in Mongolian and Han patients with non-dipping hypertension.Methods A total of 68 cases of Mongolian with non-dipping hypertension who had settled in Inner Mongolia and had not intermarried with other nationalities within three generations and 70 cases of Han patients with non-dipping hypertension were consecutively selected as research object,they all received diagnosis at the Second Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science and Technology from September 2022 to September 2023,and were divided into two groups on average according to the simple randomization method,the group of medication after dinner(group A)and the group of medication before bedtime(group B),and both groups took irbesartan 75mg in the morning,once a day.After that amlodipine benzenesulfonate tablets 5mg,once a day,were added after dinner in group A and before bedtime in group B.Treatment was carried out for 6 months.The ambulatory blood pressure parameters of total patients,Han patients and Mongolian patients before and after treatment were compared between two groups,as well as the proportion of nonpareil blood pressure and the blood pressure compliance rate of total patients after treatment between two groups.Results ①Dynamic blood pressure indicators:After treatment,the dynamic blood pressure levels in both groups of patients decreased,and the decrease in indicators in group A was more significant than that in group B(P＜0.05);Han and Mongolian patients were compared within their respective populations,and the results were consistent with the inter group comparison of the total patients(P＜0.05).②Correction of non dipper blood pressure:After treatment,the effective rates of dipper blood pressure in group A were higher than those in group B,and the difference was statistically significant(P＜0.05).③Blood pressure compliance rate:The blood pressure compliance rate of group A patients after treatment was higher than that of group B,and the difference was statistically significant(P＜0.05).Conclusion Amlodipine benzenesulfonate taken at different times has certain therapeutic effects on patients with non-dipping hypertension,and therapeutic effects are better compared with the effect of patients taking the drug after dinner,the degree of improvement of ambulatory blood pressure indexes,correction rate of non-dipping hypertension and blood pressure compliance rate are better than those of patients who take the drug before bedtime,and the effect of taking the drug after dinner is better than that take the drug before bedtime for both Mongolian and Han patients with non-dipping hypertension.

Objective:To investigate the changes in plasma ghrelin and influencing factors of weight loss effects after laparoscopic sleeve gastrectomy combined with fundoplication surgery (LSGFD).Methods:The retrospective cohort study was conducted. The clinical data of 115 obesity patients who were admitted to the People′s Hospital of Xinjiang Uygur Autonomous Region from April to June 2023 were collected. There were 37 males and 78 females, aged (37±9)years. Of 115 pati-ents, 93 cases undergoing laparoscopic sleeve gastrectomy (LSG) were divided into the LSG group, and 22 cases undergoing LSGFD were divided into the LSGFD group. Measurement data with normal distribution were represented as Mean± SD, and the independent sample t test was used for com-parison between groups. Count data were described as absolute numbers, and the chi-square test was used for comparison between groups. Repeated measurement data were analyzed using the repeated ANOVA, and their variances were tested using a spherical test. The Logistic regression model was used for univariate and multivariate analyses. Results:(1) Changes in preoperative and postoperative plasma ghrelin in two groups of patients. The plasma ghrelin of patients at preopera-tive and postoperative 6 months changed from (16±14)×10 2 ng/L to (10±4)×10 2 ng/L in the LSG group and changed from (12±11)×10 2 ng/L to (11±3)×10 2 ng/L in the LSGFD group. There was no significant difference in the time effect, inter group effect, and interaction effect of changes in plasma ghrelin between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=2.199, Fgroup=0.001, Finteraction=0.793, P>0.05). There was a significant difference in plasma ghrelin in the LSG group before and after surgery at 6 months ( t=4.148, P<0.05), and there was no significant difference in plasma ghrelin in the LSGFD group before and after surgery at 6 months ( t=0.622, P>0.05). (2) Changes in preoperative and postoperative weight loss and metabolic related indicators in two groups of patients. ① There was a significant difference in the time effect of changes in body mass between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=242.285, P<0.05), and there was no significant difference in the inter group effect and interaction effect of changes in body mass between the LSG group and the LSGFD group before and after surgery at 6 months ( Fgroup=1.163, Finteraction=0.606, P>0.05). There were significant differences in body mass in the LSG group or the LSGFD group before and after surgery at 6 months ( t=23.597, 14.680, P<0.05). ② There was a significant difference in the time effect of changes in body mass index (BMI) between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=382.431, P<0.05), and there was no significant difference in the inter group effect and interaction effect of changes in BMI between the LSG group and the LSGFD group before and after surgery at 6 months ( Fgroup=1.619, Finteraction=1.085, P>0.05). There were significant differences in BMI in the LSG group or the LSGFD group before and after surgery at 6 months ( t=25.645, 16.628, P<0.05). ③ There was a significant difference in the time effect of changes in excess weight loss (%EWL) between the LSG group and the LSGFD group after surgery at 1 to 6 months ( Ftime=666.136, P<0.05), and there was no significant difference in the inter group effect and interaction effect of changes in %EWL between the LSG group and the LSGFD group after surgery at 1 to 6 months ( Fgroup=0.127, Finteraction=0.498, P>0.05). ④ There was no significant difference in the time effect, inter group effect, and interaction effect of changes in fasting blood glucose between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=1.573, Fgroup=1.872, Finteraction=0.948, P>0.05). There was a significant difference in fasting blood glucose in the LSG group before and after surgery at 6 months ( t=2.675, P<0.05), and there was no significant difference in fasting blood glucose in the LSGFD group before and after surgery at 6 months ( t=1.074, P>0.05). ⑤ There were significant differences in the inter group effect and interaction effect of changes in triglyceride between the LSG group and the LSGFD group before and after surgery at 6 months ( Fgroup=8.419, Finteraction=3.180, P<0.05), and there was no significant diffe-rence in the time effect of changes in triglyceride between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=1.398, P>0.05). Results of individual effect shown that there was no significant difference in triglyceride in the LSG group or the LSGFD group before and after surgery at 3 months ( F=2.956, 3.248, P>0.05), and there were significant differences in trigly-ceride in the LSG group or the LSGFD group after surgery at 1 month and 6 months ( F=14.152, 3.477, P<0.05). There was a significant difference in triglyceride in the LSG group before and after surgery at 6 months ( t=3.164, P<0.05), and there was no significant difference in triglyceride in the LSGFG group before and after surgery at 6 months ( t=0.023, P>0.05). ⑥ There were significant differences in the time effect and inter group effect of changes in total cholesterol between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=3.662, Fgroup=7.591, P<0.05), and there was no significant difference in the interaction effect of changes in total cholesterol between the LSG group and the LSGFD group before and after surgery at 6 months ( Finteraction=0.626, P>0.05). There was a significant difference in cholesterol in the LSG group before and after surgery at 6 months ( t=3.253, P<0.05), and there was no significant difference in total cholesterol in the LSGFG group before and after surgery at 6 months ( t=1.567, P>0.05). ⑦ There were significant differences in the time effect and inter group effect of changes in uric acid between the LSG group and the LSGFD group before and after surgery at 6 months ( Ftime=15.306, Fgroup=4.244, P<0.05), and there was no significant difference in the interaction effect of changes in uric acid between the LSG group and the LSGFD group before and after surgery at 6 months ( Finteraction=0.968, P>0.05). There were significant differ-ences in uric acid in the LSG group or the LSGFG group before and after surgery at 6 months ( t=6.152, 3.660, P<0.05). (3) Analysis of influencing factors on postoperative weight loss effects. Results of multivariate analysis showed that preoperative BMI, postoperative 6 months plasma ghrelin were independent protective factors for postoperative weight loss effects ( odds ratio=0.881, 0.673, 95% confidence interval as 0.817-0.950, 0.577-0.787, P<0.05). Conclusions:The decrease in plasma ghrelin in patients after LSGFD is not as obvious as that in patients after LSG, but it can achieve the same weight loss and metabolic improvement effects as after LSG. The lower preoperative BMI and postoperative 6 months plasma ghrelin are independent protective factors for postoperative weight loss effects.

AIM:To investigate the impact of honokiol(HKL),an activator of silent information regulator 3(SIRT3),on postoperative cognitive dysfunction(POCD)in mice,and to explore the potential mechanisms.METHODS:Ten-month-old male C57/BL6 mice were randomly divided into control(Con)group,surgical(Sur)group and Sur+HKL group(n=10).The mice in Sur+HKL group were intraperitoneally injected with HKL for 7 d before modeling.The mice in Sur and Sur+HKL groups underwent tibial fracture open reduction and internal fixation to establish the POCD model.The assessment of cognitive function was conducted using the open-field test(OFT),novel object recognition test(NORT),Morris water maze test(MWMT),and Y-maze test(YMT).Nissl staining was employed to assess the morphology,struc-ture and vitality of hippocampal and cortical neurons in mice.The protein expression of glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),acyl coenzyme A synthetase long-chain family member 4(ACSL4),SIRT3 and nuclear factor E2-related factor 2(NRF2)in the mouse hippocampus was detected by Western blot,while im-munofluorescence staining was utilized to determine GPX4 level in mouse neurons.RESULTS:No statistically signifi-cant differences were observed among the groups in terms of total distance moved and central zone exploration during the OFT(P>0.05).However,the results from the NORT and YMT indicated that the mice in Sur group exhibited significant-ly lower recognition indexes,reduced alternation rates(P<0.01),and decreased percentages of entries and crossing time into the new arm after side arm blockade(P<0.01),when compared with Con group.Furthermore,the mice in Sur group demonstrated a slower decrease in latency during the learning period of MWMT,while significantly lower latency,fewer crossing number and lower percentage of time in the target quadrant were observed during the testing period of MWMT(P<0.01).The above indicators were obviously enhanced in Sur+HKL group compared with Sur group(P<0.01).The results of Nissl staining indicated lighter neuronal staining in the hippocampal CA1 region and medial prefrontal cortex in Sur group,accompanied by a significant reduction in the number of Nissl-stained positive neurons(P<0.01).Notably,HKL pretreatment demonstrated a significant improvement in neuronal vitality.Analysis of Western blot revealed that compared with Con group,the expression of SIRT3,GPX4,SLC7A11 and NRF2 in Sur group was significantly reduced,while the expression of ACSL4 was significantly increased(P<0.05).However,these alterations were reversed after treatment with HKL(P<0.05).Immunofluorescence staining of hippocampal neurons corroborated the findings from Western blot analy-sis,demonstrating a notable decrease in GPX4 expression in hippocampal neurons of Sur group,which was significantly restored after HKL pretreatment(P<0.01).CONCLUSION:Treatment with HKL attenuates POCD in mice,potentially through its inhibitory effect on hippocampal neuronal ferroptosis.

ObjectiveTo analyse the current implementation status of Chinese herbal medicine （CHM） placebo and systematically evaluate the placebo effect in randomised controlled trials （RCTs） of traditional Chinese medicine （TCM） for the treatment of functional dyspepsia （FD）. MethodsA combination of medical subject terms and free words was used to search six databases， including PubMed， EMBASE， Cochrane Library， Web of Science， China National Knowledge Infrastructure， and Wanfang， for RCTs with CHM placebo group for FD published from January 31st， 1994 to September 30th， 2023. The dosage forms， composition， and methodological quality were collected and evaluated. The quality of the included articles was evaluated by Cochrane risk of bias assessment tool， and meta-analysis was performed on the CHM placebo response rate of patients with FD， and subgroup analysis and meta-regression was performed according to diagnostic criteria， efficacy criteria， duration of treatment， type of placebo， whether it contained active ingredient， and whether it evaluated placebo effects. ResultsA total of 34 publications were included involving 5046 participants， of which 2221 FD patients received CHM placebo treatment. Granules were the predominant placebo preparation， accounting for 71% （24/34）； 32.35% （11/34） of the studies added real CHM to the placebo， and only 12 （35%） of the studies described appearance， odour， and taste. The placebo response rate in FD patients in the placebo group was 41% （95% CI： 0.35 to 0.47； P＜0.01， I² = 87%）； there was significant difference between groups with different diagnostic criteria and different treatment durations （P＜0.05 or P＜0.01）， but there was no significant difference between the different efficacy evaluation criteria， the different placebo preparation， the presence of a low-dose active ingredient， and the presence or absence of placebo assessment （P＞0.05）. ConclusionThere was a significant CHM placebo effect in patients with FD， with granules as the main preparation of placebop. Different diagnostic criteria and different treatment times may affect the response rate of patients， and the addition of low-dose real medicine to the CHM placebos has not been seen to have an effect on the response rate. Clinical investigators have not paid enough attention to placebos， and there is a lack of uniform standards and norms for the preparation and evaluation of CHM placebos.

ObjectiveTo investigate the possible mechanism of modified Gegen Qinlian Decoction （葛根芩连汤） in treatment of ulcerative colitis （UC） from the view of intestinal mucosal epithelial barrier damage and epithelial mesenchymal transition. MethodsSixty male C57BL/6J mice were divided into blank group， model group， western medicine control group， and low-， medium-， and high-dose modified Gegen Qinlian Decoction groups， with 10 mice in each group. Except for the blank group， 3% dextran sodium sulfate （DSS） was used to induce colitis model by free drinking for 7 days， and on the first day of modelling， 6， 12， and 24 g/（kg·d） of modified Gegen Qinlian Decoction were given to the low-， medium-， and high-dose groups respectively， 5-aminosalicylic acid （5-ASA） 100 mg/（kg·d） given by gavage to western medicine control group， and 10 ml/kg distilled water were given to blank and model group by gavage， once a day for 7 days. Body mass of mice was recorded and disease activity index （DAI） scores were performed daily. The mice were anesthetized after 24h of the last administration and the colon was taken to observe the length of colon， HE staining was applied to observe the damage of colonic mucosa and score pathological states， Masson staining to detect the deposition of colonic collagen fibers， immunofluorescence to observe the distribution of F-actin in colonic mucosal epithelium， and immunohistochemistry to detect the expression of tight junction protein ZO-1， Occludin， E-cadherin and Vimentin. ResultsCompared with the blank group at the same time， the percentage of body mass of mice in the model group on day 7 of modelling significantly reduced and the DAI score was significantly increased （P＜0.01）； compared with the model group at the same time， the body mass of mice in the western medicine control group and all of modified Gegen Qinlian Decoction groups decreased， and the DAI scores of mice in the western medicine control group and the high-dose modified Gegen Qinlian Decoction group decreased （P＜0.05 or P＜0.01）； compared with the same time of mice in the low-dos Gegen Qinlian Decoction group， the body mass of mice in the high-dose Gegen Qinlian Decoction group and the western medicine control group significantly elevated （P＜0.05）. Compared with the blank group， the length of the colon of mice in the model group was significantly shortened， the pathological score and the percentage of collagen area were significantly increased， the average fluorescence intensity of F-actin was reduced， the protein levels of ZO-1， Occludin and E-cadherin in the colon tissue decreased， and the protein level of Vimentin elevated （P＜0.01）. Compared with the model group， the length of colon significantly increased， patholo-gical score， collagen area percentage decreased， ZO-1， Occludin， E-cadherin protein levels increased and Vimentin levels decreased in all medicated groups； the average fluorescence intensity of F-actin increased in the western medicine control group and the middle- and high-dose Gegen Qinlian Decoction groups （P＜0.05 or P＜0.01）. Compared with the low-dose Gegen Qinlian Decoction group， the proportion of collagen fibre area in the middle-dose Gegen Qinlian Decoction group and the western medicine control group reduced； the mean fluorescence intensity of F-actin increased in the middle-dose Gegen Qinlian Decoction group； the protein levels of ZO-1 and E-cadherin increased in the western medicine control group， and the protein levels of ZO-1 increased in the high-dose Gegen Qinlian Decoction group （P＜0.05）. Compared with the medium-dose Gegen Qinlian Decoction group， the protein levels of ZO-1 elevated in the high-dose Gegen Qinlian Decoction group （P＜0.05）. Comapred with the high-dose Gegen Qinlian Decoction group， level of E-cadherin and Vimentin protein of the western medicine control group increased （P＜0.05）. ConclusionModified Gegen Qinlian Decoction was able to reduce colonic inflammation and mucosal barrier damage and inhibit the process of epithelial mesenchymal transition in mice models of ulcerative colitis， which may be one of its action mechanisms .

OBJECTIVE To study the chemical composition and the function of solid excipients in relieving the "nourishing and spleen-impairing" effect of Zhangbang nine steaming nine sun-dying Rehmanniae Radix(RR) by using UPLC-Q-TOF-MS technique. METHODS UPLC-Q-TOF-MS was used to collect the data of Zhangbang nine steaming nine sun-dying RR, and UNIFI platform and self-built database were used for the rapid qualitative analysis and identification of the chemical components of Zhangbang nine steaming nine sun-dying RR. The effect of solid excipients on the function of RR "nourishing and spleen-impairing" was investigated by establishing a mouse model of low gastric motility and measuring its gastric residual rate and small intestine propulsion rate. RESULTS The UPLC-Q-TOF-MS and UNFI platforms identified 76 chemical components in nine steaming nine sun-dying RR, including 9 iridoid glycosides, 11 phenylethyl glycosides, 31 flavonoids, 6 saccharides and 19 other compounds; the results of gastric emptying and small intestinal propulsion experiments on mice showed that Zhangbang nine steaming nine sun-dying RR had gastric emptying and intestinal propulsion effects. CONCLUSION This method can be used for the rapid identification and analysis of nine steaming nine sun-dying RR, which has solid excipients Citri Reticulatae Pericarpium and Amomi Fructus to alleviate the "nourishing and spleen-impairing" function of RR. This study can provide a reference for quality control and elucidation of the material basis of the medicinal effects of specialty beverage of Zhangbang nine steaming nine sun-dying RR.

ObjectiveTo assess the capacity of health emergency drills for poisoning emergencies at the municipal level in Guangdong Province. Methods A total of 21 municipal teams from cities in Guangdong Province participated in the health emergency drill competition, which included comprehensive tests and practical assessments. Results The pass rate for the total score, comprehensive tests, practical assessments of 21 municipal teams was 66.7%, 33.3%, 66.7%, respectively. The pass rate of the comprehensive tests was lower than that of practical assessments (P<0.01). The pass rate for the total score, comprehensive tests, and practical assessments of team from the Pearl River Delta region was higher than those in non-Pearl River Delta regions (88.9% vs 50.0%, 55.5% vs 16.7%, 88.9% vs 50.0%). For the four comprehensive test items, the highest pass rate was for personal protective principles against chemical poisoning (57.1%). For the five practical assessment items, the highest pass rate was for the selection and matching of personal protective equipment and practice of poisoning detection (both 71.4%). Conclusion It is urgent to improve the capacity of health emergency drills at the municipal level in Guangdong Province. Emphasis should be placed on strengthening capacity building in teams from non-Pearl River Delta regions.

Objective To investigate the expression of Midkine(MDK)in cholangiocarcinoma(CCA)and its value in predicting the prognosis of CCA,as well as the potential mechanism of the effect of MDK on the progression of CCA.Methods The data of CCA samples were obtained from TCGA database to analyze the difference in the expression of MDK between cancer tissue and paracancerous tissue and its association with clinical features,and the data collected from GEO database and 11 CCA patients who underwent surgical resection in The Fifth Medical Center of Chinese PLA General Hospital from June 2018 to September 2021 were used for validation.STRING and Cytoscape were used to construct a protein-protein interaction network,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were used to investigate the biological functions and tumor-related pathways involving MDK-related genes.In addition,TIMER and TISIDB databases were used to analyze the correlation between MDK expression and immune cell infiltration in CCA tissue.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the Fisher's exact test was used for comparison of categorical data between two groups.The Kaplan-Meier method was used to plot survival curves,and the Log-rank test was used for comparison between groups.The Spearman correlation analysis was used to investigate the correlation between two variables.Results The expression level of MDK in cancer tissue and paracancerous tissue of CCA patients was compared based on TCGA database,and the results of the non-paired and paired analyses showed that the expression level of MDK in CCA tumor tissue was significantly higher than that in paracancerous tissue(P<0.001).Transcriptome sequencing was performed for the tumor tissue and its corresponding paracancerous tissue from 11 CCA patients,and the results showed that the expression level of MDK in CCA tumor tissue was significantly higher than that in corresponding paracancerous tissue(P<0.01).High expression of MDK was associated with lymph node metastasis(P=0.045)and vascular invasion(P=0.044).Survival analysis showed that compared with the CCA patients with low MDK expression,the CCA patients with high MDK expression had significantly shorter overall survival time(χ2=5.30,P=0.028)and disease-specific survival time(χ2=6.25,P=0.019).The GO and KEGG enrichment analyses showed that the 30 MDK-related genes were closely associated with ubiquitin-mediated proteolysis and affected the prognosis of CCA patients.The TIMER analysis showed that the expression level of MDK was positively correlated with the infiltration of B cells(r=0.356,P=0.035 6)and dendritic cells(r=0.409,P=0.014 7)in tumor microenvironment of CCA;the TISIDB analysis showed that the expression level of MDK was positively correlated with CXCL16(r=0.465,P=0.004 67)and was negatively correlated with CXCL12(r=-0.389,P=0.019 7)and CXCR5(r=-0.393,P=0.018 5),and it was also negatively correlated with the immune checkpoint regulators VTCN1(r=-0.393,P=0.018 3),LTA(r=-0.380,P=0.022 7),and PVR(r=-0.350,P=0.037 3).Conclusion High expression of MDK is associated with poor prognosis in CCA patients,and MDK has the potential of being used as a molecular marker for predicting the prognosis of CCA.MDK may promote the development and progression of CCA by regulating ubiquitin-mediated proteolysis and the infiltration of B cells and dendritic cells.