Objective To investigate the effects of menaquinone-4(MK4)on the activation and function of CD8+T cells.Methods An in vitro culture system for primary mouse CD8+T cells was established by isolating these cells from the spleen using CD8a T cell isolation kit.The isolated CD8+T cells were then incubated and activated in a 96-well plate coated with anti-CD3/CD28 antibodies.The impact of MK4 on the activation and cytokine secretion of CD8+T cells was explored by quantifying the expression levels of CD8+T cell activation receptors and cytokines using flow cytometry.Additionally,the concentrations of these cytokines in the culture supernatant were measured by enzyme-linked immunosorbent assay(ELISA).The influence of MK4 on the anti-tumor function of CD8+T cells was evaluated by co-culturing colorectal cancer MC38 cells of mice with CD8+T cells at different ratios,and the effect of MK4 was assessed by detecting tumor cell apoptosis.Results High-purity primary CD8+T cells of mice(97.5％)were isolated using the magnetic bead,which could be activated by pre-coated CD3/CD28 antibodies and showed proliferation.Compared with the control group,the MK4-treated group exhibited increased expressions of CD25,CD69 and CD44 on CD8+T cells,as well as higher production and secretion levels of interleukin-2(IL-2),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)and granzyme B.In addition,CD8+T cells in the MK4-treated group induced a higher percentage of tumor cell apoptosis(36.7％)compared with the control group(29.1％).Conclusion MK4 can enhance the activation of CD8+T cells and promote anti-tumor functions.

Objective To investigate the clinical value of indocyanine green (ICG) fluorescence navigation in laparoscopic cholecystectomy (LC) after percutaneous transhepatic gallbladder drainage (PTGBD) in elderly patients with acute cholecystitis. Methods A retrospective analysis was performed for the perioperative clinical data of the elderly patients with acute cholecystitis who underwent LC after PTGBD in Zhengzhou Central Hospital Affiliated to Zhengzhou University from June 2021 to February 2022. The 36 patients who underwent ICG fluorescence navigation were enrolled as experimental group, and the 26 patients who did not undergo ICG fluorescence navigation during the same period of time were enrolled as control group. Preoperative general information was analyzed for both groups, as well as time to identify the biliary system during surgery, time of operation, intraoperative blood loss, bile tract injury, conversion to laparotomy, time to first flatus after surgery, time to ambulation, time to removing abdominal drainage tube, time to return to normal diet, and length of postoperative hospital stay. The group t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between groups. Results Compared with the control group, the experimental group had a significantly shorter time to identify the biliary system during surgery (19.9±3.7 min vs 36.5±5.9 min, t =13.56, P < 0.05), a significantly shorter time of operation (50.6±8.5 min vs 80.9±10.6 min, t =12.48, P < 0.05), and a significantly lower amount of intraoperative blood loss ( χ 2 =6.91, P < 0.05). No patient was converted to laparotomy in the experimental group, while 2 patients in the control group were converted to laparotomy, and no bile duct injury was observed in either group. Compared with the control group, the experimental group had significantly shorter time to ambulation (10.2±2.4 hours vs 16.6±3.2 hours, t =8.92, P < 0.05), time to removing abdominal drainage tube (20.1±3.4 hours vs 30.7±4.7 hours, t =10.2, P < 0.05), time to return to normal diet (20.3±3.8 hours vs 31.2±6.0 hours, t =8.68, P < 0.05), and length of postoperative hospital stay [3.3 (3.0-4.3) days vs 5.3 (5.0-6.2) days, Z =5.91, P < 0.05]. Conclusion ICG fluorescence navigation can visualize the extrahepatic biliary system during LC after PTGBD in elderly patients with acute cholecystitis, which may help to achieve accurate operation, reduce the risk of surgery, shorten the time of operation, and accelerate postoperative recovery.

ObjectiveTo establish a neuroinflammation-based obesity and depression comorbidity （COM） model in mice and explore the pharmacodynamics and preliminary pharmacological mechanism of tripterine on COM mice. MethodC57BL/6J mice were randomly divided into a normal group （Chow）， a diet-induced obesity group （DIO）， and a COM group. The mice in the COM group were fed on a high-fat diet and chronically stressed with moist litter for 12 weeks to establish the COM model. C57BL/6J mice were randomly divided into a Chow group， a COM group， and a tumor necrosis factor-α（TNF-α） knock-down group. In the TNF-α knock-down group， TNF-α shRNA adeno-associated virus was injected into the amygdala through brain stereotaxis， and the expression of TNF-α in the amygdala was down-regulated. C57BL/6J mice were randomly divided into a Chow group， a DIO group， a DIO + low-dose tripterine group （0.5 mg·kg^-1）， a DIO + high-dose tripterine group （1.0 mg·kg^-1）， a COM group， a COM + low-dose tripterine group （0.5 mg·kg^-1）， and a COM + high-dose tripterine group （1.0 mg·kg^-1）. The body weight， food intake， glucose tolerance， white/brown fat ratio， serum total cholesterol （TC）， triglyceride （TG）， and high-/low-density lipoprotein cholesterol （HDL-C and LDL-C） content were recorded， and obesity of mice in each group was evaluated. Forced swimming test （FST）， tail suspension test （TST）， and open field test were used to evaluate the degree of depression of mice in each group. Immunofluorescence staining was used to detect the protein expression levels of neuropeptide Y， tryptophan hydroxylase 2 （TPH2）， and brain-derived neurotrophic factor （BDNF） in various brain nuclei of mice. Correlation analysis was used to detect the correlation of obesity and depression indexes. ResultThe comparison of the Chow group and the DIO group indicated that COM mice showed obesity and depression. To be specific， obesity was manifested as increased body weight and food intake （P<0.05， P<0.01）， as well as increased NPY expression in the central amygdala， and depression was manifested as prolonged immobility time in FST and TST （P<0.01）， and reduced TPH2-positive 5-hydroxytryptamine neurons in the dorsal raphe nucleus （DRN） and basolateral nucleus of the amygdala （BLA）. The down-regulation of TNF-α protein in BLA of COM mice shortened the immobility time in FST and TST （P<0.05， P<0.01）， increased TPH2/BDNF-positive neurons in BLA， and showed no significant changes in obesity. In DIO mice， the administration of 0.5 mg·kg^-1 tripterine for 9 days significantly decreased the 60 min blood glucose in glucose tolerance （P<0.01） and food intake （P<0.05）. In COM mice， 1.0 mg·kg^-1 tripterine was administered for 14 days to significantly decrease 30 min blood glucose in glucose tolerance （P<0.01）， and food intake （P<0.05）， and immobility time in TST （P<0.01）， increase TPH2-BDNF double-labeled cells in BLA and DRN， and reduce the area of TMEM119-stained cells. ConclusionThe model of obesity and depression comorbidity can be properly induced in mice under the condition of dual stress of energy environment. Tripterine can effectively interfere with obesity-depression comorbidity， and its mechanism may be related to the inhibition of central nervous system inflammation.

The complexity of oral ulcerations poses considerable diagnostic and therapeutic challenges to oral specialists. The expert consensus was conducted to summarize the diagnostic work-up for difficult and complicated oral ulcers, based on factors such as detailed clinical medical history inquiry, histopathological examination, and ulceration-related systemic diseases screening. Not only it can provide a standardized procedure of oral ulceration, but also it can improve the diagnostic efficiency, in order to avoid misdiagnosis and missed diagnosis.
Consensus ; Humans ; Oral Ulcer/therapy*

Objective:To explore the effect of key amino acid mutations of 3D protein in enterovirus 71 on viral proliferation, and infer the mechanism of mutation affecting viral proliferation according to the tertiary structure model of 3D protein.Methods:The proliferation characteristics of the mutant strain that was constructed by site-directed mutagenesis and reverse genetics using the pMD19T-SDLY107-EGFP constructed from the fatal strain SDLY107 as a template were measured. Meanwhile the tertiary structure of the 3D protein was predicted, and then the possible mechanism of the mutation affecting the proliferation ability of the virus was speculated according to the functional characteristics of the domain of the 3D protein.Results:Two mutant viruses, eGFP-EV-A71 (S37N) and eGFP-EV-A71 (R142K), were successfully constructed by double enzyme digestion and viral gene sequencing. The proliferation rate of the two mutant strains in RD cells was significantly lower than that of the parent strains. The 3D protein tertiary structure prediction model showed that the 3D protein consisted of three domains: "Finger" , "thumb" and "Palm" , constituting a cupped right-handed structure. S37N and R142K are located in the "thumb" domain and " finger" domain, respectively. The "thumb" and "finger" domains have important effects on the activity of 3D polymerase and the stability of protein.Conclusions:Mutations at S37N and R142K sites of EV71 3D protein decrease the replication ability of EV71, and these two mutations may affect the proliferation of EV71 virus by changing the 3D protein polymerase activity and the interactions between multiple domains.

Objective:We try to screen out predictive indicators with higher value by analyzing the differences in clinical and laboratory indicators between severe fever with thrombocytopenia syndrome (SFTS) patients in the intensive care unit (ICU) group and non-ICU group.Methods:The clinical and laboratory index data of 69 SFTS patients diagnosed in the laboratory in a hospital from June to December 2019 were retrospectively collected. According to the clinical outcome of the patients, they were divided into ICU and non-ICU groups. The differences in clinical manifestations and laboratory indicators between the two groups were analyzed. The receiver operating characteristic curve (ROC) was used to screen the more valuable predictive indicators.Results:Compared with the non-ICU group, ICU group SFTS patients had significantly higher procalcitonin (PCT), C-reactive protein (CRP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glutamyl transpeptidase (GGT), leucine aminopeptidase (LAP), glutamate dehydrogenase (GDH), adenosine deaminase (ADA), cystatin C (Cys C), α-hydroxybutyrate dehydrogenase (α-HBDH), creatine kinase (CK), lactate dehydrogenase (LDH) levels ( W=530.0, P=0.003; W=496.5, P=0.015; W=496.0, P=0.015; W=535.5, P=0.002; W=545.5, P=0.001; W=498.5, P=0.013; W=537.0, P=0.002; W=523.0, P=0.004; W=512.0, P=0.007; W=502.0, P=0.012; W=486.0, P=0.023; W=509.0, P=0.008; W=541.0, P=0.002) and significantly lower platelet count (PLT), indirect bilirubin (IBIL), albumin/globulin ratio(A/G) and superoxide dismutase (SOD) levels ( W=199.0, P=0.024; W=175.5.5, P=0.009; t=-2.9, P=0.004; W=209.5, P=0.036; t=-3.0, P=0.004). ROC result showed that ALP [area under the curve (AUC)=0.804, 95% confidence interval ( CI) (0.679~0.929)] and LDH [AUC=0.805, 95% CI (0.680~ 0.930)] have a higher value for predicting the risk of severe illness. Conclusions:Abnormal liver function, heart function, and renal function indicators in SFTS patients indicate that patients are at risk of exacerbation. Among them, ALP and LDH levels have higher predictive value for risk of severe disease, suggesting that the monitoring of patients with the above symptoms should be strengthened in the clinical nursing process.

Objective: To investigate the development of public health human resource of the Centers for Disease Control and Prevention (CDCs) in Zhejiang Province from 2016 to 2020, so as to put forward the strategies for the talent team construction of CDCs. Methods: The general information of personnel of CDCs in Zhejiang Province from 2016 to 2020 was collected through China Information System for Disease Control and Prevention. A questionnaire survey and a qualitative interview were conducted to collect the data of staffing, talent training, salary and employee turnover, and to analyze the development of the talent team of CDCs. Results: By 2020, there were 105 CDCs, including 1 provincial, 11 prefectural and 93 county-level CDCs, and 5 277 staff on-the-job. From 2016 to 2020, the number of staff per 10 000 permanent residents in the province, prefecture and county levels were 0.062-0.070, 0.170-0.188 and 0.585-0.604, respectively, which did not meet the standard for the personnel establishment in Zhejiang CDCs; the proportion of the staff with bachelor degree or above increased from 90.05% to 94.25%; the proportion of the health professionals decreased from 82.91% to 80.01%; the proportion of the staff with senior professional titles increased from 35.20% to 45.25%. The results of the qualitative interview indicated a lack of health professionals and high-end talents, a gap between the approved staffing and post requirements, as well as demands for a stable team and higher pay. Conclusion From 2016 to 2020, the structure of educational background, major and professional title among the on-the-job staff of CDCs in Zhejiang Province has been improving; however, the total allocation of human resource is insufficient, and the stability and treatment of staff need to be further improved.

Objective:To investigate the diagnosis and treatment of hepatic artery thrombosis (HAT) after adult orthotopic liver transplantation.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 411 patients who underwent adult orthotopic liver transplantation in the First Affiliated Hospital of Xi ′an Jiaotong University from December 2011 to July 2018 were collected. There were 328 males and 83 females, aged from 21 to 66 years, with a median age of 46 years. Observation indicators: (1) incidence of HAT and its clinical characteristics; (2) diagnosis of HAT; (3) treatment of HAT; (4) follow-up. Follow-up using outpatient service, telephone interview or WeChat group communication was conducted to detect the incidence of biliary stricture and survival of patients up to August 2018. Measurement data with normal distribution were represented as Mean± SD, measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers or percentages. Survival rate was estimated using the Kaplan-Meier method. Results:(1) Incidence of HAT and its clinical characteristics: 11 of 411 patients had HAT after orthotopic liver transplantation with the incidence of 2.68%(11/411), including 10 males and 1 female, aged 44 years(range, 22-63 years). The time to occurrence of postoperative HAT was 4 days(range, 1-15 days). The etiologies of 11 patients included 6 cases of hepatitis B virus-related cirrhosis, 1 case of hapatitis related cirrhosis, 1 case of hepato-cellular carcinoma, 1 case of liver cirrhosis, 1 case of alcoholic hepatitis related cirrhosis, 1 case of wilson disease. All the 11 patients were ABO compatible. The cold ischemic time and warm ischemic time of donor liver were (316±89)minutes and (13±4)minutes, respectively. Type Ⅰ arterial anasto-mosis was conducted in 11 patients. The clinical manifestations included asymptomatic type in 10 patients and sepsis type in 1 patient. (2) Diagnosis of HAT: all the 11 patients were confirmed with HAT by endovascular angiography, including 7 cases showed no arterial flow under Color Doppler ultrasound, and contrast-enhanced ultrasound indicated HAT. Two patients showed increased hepatic artery resistance index under Color Doppler ultrasound, and contrast-enhanced ultrasound indicated 1 case of HAT and 1 case of anastomotic stenosis. One patient showed slow velocity of hepatic artery blood flow and low resistance index under color Doppler ultrasound, and contrast-enhanced ultrasound indicated HAT. One patient showed slight blood flow signals under Color Doppler ultrasound, and contrast-enhanced ultrasound indicated HAT. (3) Treatment of HAT: 11 patients received endovascular therapy. Six patients had HAT completely disappeared after thrombolytic therapy, 5 patients with residual thrombosis continued thrombolytic therapy with microcatheter urokinase. Six patients with complications were improved after symptomatic treatment. HAT completely disappeared after (6.7±2.6)days of treatment and the clinical success rate was 11/11. (4) Follow-up: 11 patients were followed up for 19-1 722 days, with a median follow-up time of 46 days. During the follow-up, 4 patients had biliary stricture and underwent stent implantation. Nine patients survived with 1-, 3-, 5-year overall survival rates of 75%, 75%, 75%, and 2 patients died.Conclusions:The incidence of HAT after adult orthotopic liver transplantation is low and clinical manifestations are atypical. Contrast enhanced ultrasound can improve diagnosis of suspected thrombosis. Endovascular therapy is safe and effective, which can significantly improve the blood flow of hepatic artery.

Objective:To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on the maturation and differentiation of dendritic cells (DCs) and the mechanism involved in the regulation of inflammation in patients with severe acute pancreatitis (SAP).Methods:The full-term fetal umbilical cords(about 4-5 cm) were collected from Zhengzhou Central Hospital Affiliated to Zhengzhou University after cesarean section. hUC-MSCs were isolated and cultured in primary culture. Flow cytometry was used for phenotype identification, adipogenic and osteogenic staining. 20 ml peripheral blood samples from 5 SAP patients were collected, and monocytes were isolated using lymphocyte separation solution and then induced by adding granulocyte macrophage colony-stimulating factor (GM-CSF), IL-4 and tumor necrosis factor (TNF)-α and cultured as DCs. According to different culture methods, DCs were divided into DCs group, hUC-MSCs+ DCs group and hUC-MSCs+ DCs+ NS398 group (NS398 was a specific inhibitor of COX-2, a downstream regulatory gene of NF-κB). The phenotype of DCs was detected by flow cytometry, and the levels of IL-1β, IL-lα, IL-2, IL-6 and IL-10 in the supernatant of cell culture for 24 hours were determined. The expression of toll like receptor (TLR)-4, IKKα and NF-κB-p65 were detected by Western blot.Results:The hUC-MSCs were successfully cultured, and their surface markers CD 90, CD 105 and CD 73 were positively expressed, and they could differentiate into adipocytes and bone cells. With the prolongation of culture time, DCs differentiated from immature to mature cells. Compared with the DCs group, the proportion of regulatory DCs (regDCs) was increased in the hUC-MSCs+ DCs group, and the marker CD 11b was significantly up regulated [(14.26±1.25)% vs (4. 87±0.58)%], CD 1a and CD 11c were significantly down regulated [(2.81±0.34)% vs (13.62±1.52)%, (3.88±0.5)% vs (11. 8±1.22)%]. All the difference were statistically significant ( P<0.05). The expression of IL-1β, INF-γ and IL-6 in culture supernatant were down regulated, but the difference was not statistically significant; The pro-inflammatory factor IL-1α was significantly decreased [(14.91±2.58)ng/L vs (30.19±7.75)ng/L], and the anti-inflammatory factor IL-10 was significantly increased [ (17.03±4.69)ng/L vs (1.83±0.14)ng/L]. The expression levels of NF-κB-p65 and TLR4 were significantly down regulated (0.74±0.02 vs 0.97±0.01, 0.89±0.01 vs 1.72±0.01), and the expression of IKKα protein was significantly up regulated (1.12±0.01 vs 0.21±0.01) in hUC-MSCs-DCs group. All the differences were statistically significant (all P value<0.05). Compared with DCs group and hUC-MSCs+ DCs group, the expression levels of NF-κB-p65 and TLR4 were significantly down regulated (0.34±0.01 vs 0.97±0.01, 0.74±0.02 and 0.14±0.01 vs 1.72±0.01, 0.89±0.01), while the expression of IKKα protein was significantly up regulated (1.68±0.01 vs 0.21±0.01, 1.12±0.01) in hUC-MSCs+ DCS+ NS398 group. All the differences were statistically significant (all P value<0.05). Conclusions:In SAP patients, hUC-MSCs can inhibit the maturation and differentiation of DCs, and induce CD 11bhigh CD 1alow CD 11clowrregDCs to participate in immune regulation, which may play an anti-inflammatory role by inhibiting the inflammatory cascade through TLR4/IKKα/NF-κB/COX-2 pathway.

Objective:To investigate the causes,prevention and treatment of femoral artery puncture related complications caused by the application of resuscitative endovascular balloon occlusion of the aorta (REBOA) in the resection of pelvic and sacral tumors.Methods:Clinical data of 23 patients with femoral artery puncture related complications who received REBOA in the resection of pelvic and sacral tumors from August 2010 to August 2018 at the Musculoskeletal Tumor Center,Peking University People′s Hospital were retrospectively analyzed.There were 8 males and 15 females,with the age of (37.0±16.2) years (range:15 to 65 years).Arterial access via the Seldinger technique for REBOA was obtained in the right common femoral artery of 18 cases,and in the left of 6 cases.An arterial sheath with a diameter of 11 to 12 F(1 F≈0.33 mm) was used for the patient.The occurrence and treatment of postoperative complications were analyzed.Results:Acute femoral arterial thrombosis occurred in 18 patients,which was managed by open repair 48 hours postoperatively.Among the 349 patients admitted before 2015 who received hemostasis by compression after femoral artery sheath removal,12 patients (3.4%) developed acute femoral artery thrombosis.While the 476 patients admitted after 2015 who used a percutaneous suture device to close the femoral artery wound,6 patients (1.3%) developed acute femoral artery thrombosis.One case of retroperitoneal hematoma and 1 case of femoral pseudoaneurysm were found and surgically fixed.Postoperative follow-up was (40±18) months (range:13 to 108 months).Three cases with chronic lower extremity ischemia were confirmed by Doppler ultrasonography during 1 to 5 years follow-up.Two of them had minimal symptoms and denied further treatment,while the other one received femoral-femoral artery bypass surgery to restore distal flow for pain and numbness relief.Conclusions:Acute femoral arterial thrombosis is the most common femoral artery puncture.Technique refinement of REBOA,the use of percutaneous suture device and close follow-up can reduce the approach-specific complications,and help to detect and treat the complications timely,which may popularize the clinical application of REBOA.