Objective To investigate the application value of extracellular volume fraction(ECV)obtained from enhanced CT in diagnosis and classification of acute pancreatitis.Methods The clinical data from patients with acute pancreatitis(acute pancreatitis group)and normal controls(control group)underwent enhanced CT were analyzed retrospectively.The CT values of pancreas and abdominal aorta in the same sclice on precontrast and equilibrium-phase images were measured,and then pancreatic ECV was calcu-lated.The measured parameters were compared between the groups of control and acute pancreatitis,and subgroups of non-severe and severe pancreatitis.The logistic regression analysis was used to identify the risk factors for acute pancreatitis and severe pancrea-titis,and the receiver operating characteristic(ROC)curve was used to analyze the diagnostic efficiency in diagnosis and classifica-tion of acute pancreatitis.Results The pancreatic CT value and ECV were independent risk factors for acute pancreatitis(P＜0.05),and the ECV was an independent risk factor for severe pancreatitis(P＜0.05).The area under the curve(AUC)of ECV was higher in acute pancreatitis group(0.81)and severe pancreatitis subgroup(0.68).Conclusion As a quantitative parameter,the ECV obtained from enhanced CT has higher clinical application value and higher popularity in the diagnosis and classification of acute pancreatitis.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

OBJECTIVE: To compare the mRNA level of cell proliferation-related genes Twist1, SIRT1, FGF2 and TGF-β3 in placenta mesenchymal stem cells (PA-MSCs), umbilical cord mensenchymals (UC-MSCs) and dental pulp mesenchymal stem cells (DP-MSCs). METHODS: The morphology of various passages of PA-MSCs, UC-MSCs and DP-MSCs were observed by microscopy. Proliferation and promoting ability of the three cell lines were detected with the MTT method. Real-time PCR (RT-PCR) was used to determine the mRNA levels of Twist1, SIRT1, FGF2, TGF-β3. RESULTS: The morphology of UC-MSCs and DP-MSCs was different from that of PA-MSCs. Proliferation ability and promoting ability of the PA-MSCs was superior to that of UC-MSCs and DP-MSCs. In PA-MSCs, expression level of Twist1 and TGF-β3 was the highest and FGF2 was the lowest. SIRT1 was highly expressed in UC-MSCs. With the cell subcultured, different expression levels of Twist1, SIRT1, FGF2, TGF-β3 was observed in PA-MSCs, UC-MSCs and DP-MSCs. CONCLUSION Up-regulated expression of the Twist1, SIRT1 and TGF-β3 genes can promote proliferation of PA-MSCs, UC-MSCs and DP-MSCs, whilst TGF-β3 may inhibit these. The regulatory effect of Twist1, SIRT1, FGF2 and TGF-β3 genes on PA-MSCs, UC-MSCs and DP-MSCs are different.
Cell Differentiation ; Cell Proliferation/genetics* ; Cells, Cultured ; Dental Pulp/cytology* ; Female ; Fibroblast Growth Factor 2/genetics* ; Humans ; Mesenchymal Stem Cells/cytology* ; Nuclear Proteins/genetics* ; Placenta/cytology* ; Pregnancy ; Sirtuin 1/genetics* ; Transforming Growth Factor beta3/genetics* ; Twist-Related Protein 1/genetics* ; Umbilical Cord/cytology*

AIM: To explore the effect of microRNA-34a (miR-34a) on androgen receptor gene (AR) targeting regulation and proliferation and apoptosis of prostate cancer (PCa) LNCaP cells. METHODS: Thirty-six patients with PCa confirmed by prostate biopsy in urology department of our hospital were collected from October 2016 to September 2019, and 41 cases of benign prostatic hyperplasia (BPH) tissue samples were taken and operated at the same time. LNCaP cells were cultured in vitro and transfected with miR-34a mimics (mimics group), miR-34a mimic NC (NC group) respectively, and normal growth cells were set as blank control group (BC group). In addition, on the basis of mimics group, the AR over-expression vector (AR over-expression group) and its control group (AR control group) were transfected. The cell activity was detected by CCK-8 kit, the apoptosis rate was detected by Annexin V-FITC/PI double staining flow cytometry test kit, the expression of miR-34a and AR mRNA was detected by real-time quantitative PCR (RT-qPCR), the expression of AR protein, CyclinD1, c-Myc and apoptosis-related proteins (Bcl-2, Bax, capase-3) was detected by Western blotting. RESULTS: Compared with BPH tissues, the expression of miR-34a in PCa tissues decreased significantly (P<0.05), and the expression of AR mRNA and protein increased significantly (P<0.05). Compared with BC and NC group, miR-34a expression level, apoptosis rate and Bax protein expression level of LNCaP cells in mimics group increased significantly (P<0.05), the expression levels of AR, Cyclin D1, c-Myc and Bcl-2 decreased significantly (P<0.05), the activity of LNCaP cells decreased significantly at the same time (P<0.05). The dual-luciferase assay showed that miR-34a may have a direct targeting relationship with AR. Over-expression of AR gene could reverse the inhibition of miR-34a mimics on LNCaP cell proliferation and promote apoptosis. CONCLUSION: miR-34a may inhibit the proliferation and promote the apoptosis of prostate cancer LNCaP cells by regulating AR gene.

Objective: : To explore the effect of PD-1 gene knockout by CRISPR/Cas9 system on the proliferation and IFN-γ secretion in human T cells. Methods: : The sequence of sgRNA targeting PD-1 was designed. The PD-1-sgRNA and Cas9 mRNA were synthesized by T7 RNApolymerase in vitro, and then the mixture of PD-1-sgRNAand Cas9 mRNAwas delivered into activated T cells by nucleofection. The efficiency of gene knockout was confirmed by sequencing. The phenotypes of T lymphocytes and the expression of PD-1 after gene knockout were analyzed by Flow cytometry. The proliferation of T lymphocytes was calculated by trypan blue counting. The level of IFN-γ secreted by T lymphocytes was detected by ELISA. Results： ：PD-1-sgRNA and Cas9 mRNA were successfully synthesized in vitro and delivered into T cells by nucleofection. Sequencing technology confirmed that the PD-1 gene sequence was edited and the editing efficiency was 58.3%. The expression of PD-1 on T lymphocyte surface was down-regulated successfully by CRISPR/Cas9 system [(9.6±1.85)% vs (16.2±2.05)%, P<0.05]. The knockout of PD-1 gene did not affect the proliferation and phenotype of T lymphocytes(P<0.05); However, compared with the control group, the level of IFN-γ secreted by T lymphocytes in the PD-1sgRNA group was significantly increased (P<0.01). Conclusion: : CRISPR/Cas9 system can successfully ablate PD-1 gene in human T lymphocytes, which could block the negative regulation of PD-1／PD-L1 and further promote the IFN-γ secretion in T cells.

Objective To evaluate the effect of denatonium benzoaten on α-smooth muscle actin (α-SMA),subepithelial collagen and airway inflammation in asthmu mice.Methods Forty-five BALB/c mice were divided into 3 groups,normal control group (A group),asthma model group (B group),asthma model+ denatonium benzoaten group(C group);α-SMA detected by using immunohistochemistry,lung sections were stained with Masson to detect subepithelial collagen,HE stain method was used to observe the airway inflammation the images were analyzed with semi-quantitative computer.Results The deposition of α-SMA、subepithelial collagen and inflammation degree in C group was significantly reduced compared with B group,the difference were statistically significant(P＜0.05).Conclusion Denatonium benzoaten can improve airway remodeling in asthmatic mice.

Objective To analyze the correlation between the tubular gastric width and the anti gastroesophageal reflux after esophageal cancer operation,and to provide reference for the choice of surgical methods in treatment of esophageal cancer.Methods Selected 60 patients who received radical surgery for esophageal carcinoma combined with gastric tube reconstruction surgery in our hospital from January 2015 to October 2015,and divided them into two groups according to the way of stomach tube anastomosis (cervical anastomosis,thoracic anastomosis) and different width of gastric tube (greater than or equal to or less than 3 cm).Namely:cervical anastomosis + greater than or equal to 3 cm group(14 cases),cervical anastomosis + less than 3 cm group(15 cases),thoracic anastomosis + greater than or equal to 3 cm group(15 cases) and thoracic anastomosis + less than 3 cm group(16 cases).All patients recieved esophageal pH monitoring for 3 days continuously from the 11 th day after operation.The monitoring indicators include:number of reflux,accumulation time of pH ＜ 4,whether there were clinical symptoms (heartburn,chest pain,pharyngeal foreign body sensation,cough,asthma,etc.) after surgery,and the frequency and time of these clinical symptoms appeared.All the patients were given endoscopic examination at the 14th days postoperatively.Observed the esophageal mucosa of patients and conducted histopathological grading of gastric mucosal inflammation.And then made a correlation analysis of gastric tube width and esophageal mucosal inflammation grade among all the patients with reflux symptoms.Results The cumulative time and number of reflux,incidence rate of clinical symptoms,and pH values less than 4 were significantly different(P ＜ 0.05).The cumulative time and number of reflux,incidence rate of clinical symptoms,and pH values less than 4 in the cervical anastomosis + less than 3 cm group were significantly lower than that in the other 3 groups(P ＜ 0.05),with statistical significance between different groups of endoscopic esophageal mucosa inflammation grade difference (P ＜ 0.05).Esophageal mucosal inflammation grading in patients of the cervical anastomosis + less than 3 cm group was the lightest.It showed a linear correlation between the gastric tube width and esophageal mucosal inflammation grading in patients with reflux symptoms.Conclusion Postoperative gastroesophageal reflux is closely related to stomach esophagus width after resection of esophageal carcinoma with tubular stomach reconstruction of stomach esophagus,because it is unable to control gastric tube width to the appropriate range.And it should be strengthened in patients with reflux related indicators for monitoring,so as to take measures to prevent gastroesophageal reflux as soon as possible to improve the prognosis of patients with quality.

number of goblet cells, mucus secretion and mucin MUC5AC content in lung tissues. Results S100A9 in BALF of group B was (11.89±0.77) ng/mL, S100A9 integrated optical density (IOD) value in airway epithelial cells was 13.96±1.62, PAS stain area /epithelial cell area was (12.53±1.21)%, relative value of MUC5AC / NADPH was 173.91±4.29, all of the above were higher than those of group A [(6.19±0.61) ng/mL, 4.97±0.30, (1.94±0.18)%, 1];S100A9 levels, IOD of S100A9 in airway epithelial cells, PAS stain area / epithelial cell area (%), relative value of MUC5AC / NADPH in group C [(10.69±0.79) ng / ml, 11.80±0.72, (10.61±0.61)%, 94.65±1.59], group D[(9.49±0.99) ng/mL, 10.39±0.59, (8.63±0.62)%, 82.08±1.12], group E [(7.54± 0.42) ng/mL, 5.63±0.84, (4.59±0.87)%, 26.30±1.94] were lower than group B, which showed a dose-dependent reduction and the difference was statistically significant (P<0.05 or P<0.01). Conclusion DB downregulates the expression level of Ca2+-binding protein S100A9 and the mucus secretion amount of the airway goblet cells in rats.

Objective: To evaluate the clinical value of computed tomography (CT)-guided ¹²⁵I seed implantation in the treatment of patients with spinal and/or paraspinal osteolytic metastatic tumors. Methods: The radiation dose distribution was planned for 27 patients with 35 spinal and paraspinal osteolytic metastatic tumors by a treatment planning system (TPS). CT-guided ¹²⁵I seed implantation was carried out in the patients, and the quality of treatment was evaluated based on CT-imaging follow-up. Results: All the 27 patients underwent CT-guided ¹²⁵I seed implantation successfully. 12 to 50 ¹²⁵I seeds were injected into each spinal or paraspinal metastatic tumor, 39.15 on average, and the specific radioactive activity of the particles ranged from 0.60 to 0.80 mCi, 0.73 mCi on average. The minimal percentage of the dose received by 90% of the target volume (D₉₀) of the spinal and paraspinal metastatic tumors ranged from 90 to 165 Gy, 115.03 Gy on average. Among the 27 patients, 21 (77.8%) had partial remission (PR) and 6(22.2%)had stable disease (SD). The Numerical Rating Scale (NRS) scores before implantation and at postoperative 3 and 6 months were 7.81±0.74, 2.04±1.10 and 1.81±0.79, respectively, (P<0.05). The assessment of pain intensity before ¹²⁵I seed implantation and at 3 postoperative months showed obvious improvements in the patients evaluated according to the American Spinal Injury Association (ASIA) impairment scale: 12 (44.4%) patients with ASIA grade C were changed to grade D, 3 (11.1%) from grade C to grade E, 8 (29.6%) from grade D to grade E, 3 (11.1%) with a stable grade D, and 1 (3.7%)with a stablegrade C. The Karnovsky performance scale (KPS) scores before treatment and at 3 months and 6 months postoperatively were 66.30±6.88, 85.93±9.31 and 87.91±8.56, respectively (P<0.05). Their local control rate (LCR) at 3 months, 6 months and 1 year postoperatively were 100%, 92.6% and 51.9%, respectively, and the overall survival rates(OSR) were 100%, 92.6% and 55.6%, respectively. Conclusions CT-guided ¹²⁵I seed implantation can significantly relieve local pain, has advantages of less complications and higher local control rate. Therefore, it is a safe, effective and feasible treatment option for patients with spinal and paraspinal osteolytic metastatic tumors.