Objective:To assess the clinical value of nomogram based on high resolution magnetic resonance vessel wall imaging (HR-VWI) features in differentiating moyamoya disease (MMD) from atherosclerotic moyamoya syndrome (A-MMS).Methods:Eighty-four patients with digital subtraction angiography (DSA)-confirmed MMD and 73 patients with DSA-confirmed A-MMS were enrolled from Department of Medical Imaging, Affiliated Hospital of Jining Medical University from June 2020 to November 2023. All patients underwent HR-VWI. A retrospective analysis was performed on their imaging data. Univariate analysis was used to compare the differences in imaging characteristics between the two groups. Multivariate Logistic regression analysis was used to screen independent influencing factors for differentiating MMD from A-MMS and a nomogram was constructed accordingly. Receiver operating characteristic (ROC) curve and calibration curve were used to evaluate the validity and calibration of the nomogram.Results:Univariate analysis showed that age, pattern of vessel wall thickening, maximum vessel wall thickness, enhancement degree of vessel wall, vessel external diameter, ipsilateral anterior cerebral artery involvement and dot sign were statistically different between the two groups ( P<0.05). Multivariate Logistic regression analysis showed that age ( OR=6.990, 95% CI: 2.340-20.360, P<0.001), pattern of vessel wall thickening ( OR=0.066, 95% CI: 0.014-0.307, P<0.001), vessel external diameter ( OR=5.224, 95% CI: 1.672-16.324, P=0.005), ipsilateral anterior cerebral artery involvement ( OR=0.160, 95% CI: 0.038-0.679, P=0.013) and dot sign ( OR=0.081, 95% CI: 0.018-0.364, P=0.001) were independent influencing factors for differentiating MMD from A-MMS. ROC curve showed that area under the curve (AUC) of this nomogram was 0.884 (95% CI: 0.821-0.947, P<0.001), and the calibration curve showed a good fit between the predicted probability and actual probability. Conclusion:Nomogram based on HR-VWI features can effectively differentiate MMD from A-MMS.

Objective:To analyze the costs and effectiveness of five common screening modes and genetic screening for thalassemia in China in order to find the optimal way and provide evidence for the implementation of thalassemia prevention and control projects in Hunan Province.Methods:From June 2020 to April 2021, 12 971 couples from 14 cities and autonomous prefectures in Hunan Province were selected as the study population. The diagnosis of thalassemia was based on the results of genetic testing. Results of routine blood test and hemoglobin electrophoresis were collected and analyzed. The efficacy of five screening modes, at the cut-off value of <80 fl or 82 fl for the mean corpuscular volume (MCV), was analyzed by positive predictive value, negative predictive value, Jorden index and cost-effectiveness ratio. Sensitivity analysis was used to assess the feasibility of genetic screening at different costs after fixing the costs of routine blood and hemoglobin electrophoresis. The five thalassemia screening models are as follows: Mode 1: The woman had a blood routine test first. If the result was positive, the spouse required a blood routine test. If both results were positive, a thalassemia gene test should be offered to the couple. Mode 2: Both husband and wife were screened by blood routine and hemoglobin electrophoresis. If one or both of them were positive, both would be tested for thalassemia gene. Mode 3: The couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing. Mode 4: The woman was screened by blood routine and hemoglobin electrophoresis. If any one of them was positive, the woman would be tested for thalassemia gene. If the gene test result was positive, the spouse should receive thalassemia gene. Mode 5: Both spouses conducted a blood routine test. If either was positive, both would conduct hemoglobin electrophoresis test. If both were positive, both spouses should receive thalassemia gene testing. Gene testing mode: The woman would be tested for thalassemia, and her spouse would have thalassemia test too if her result was positive.Results:When using MCV<80 fl as the cut-off for diagnosing thalassemia, the Youden indices of the five prenatal screening modes in Hunan Province were 0.551, 0.639, 0.898, 0.555 and 0.356, while when using MCV<82 fl as the cut-off, the Youden indices were 0.549, 0.629, 0.851, 0.548 and 0.356. When the MCV cut-off value was <80 fl, the missed diagnosis rates of the five screening modes were 44.44%, 0.00, 0.00, 18.52% and 62.96%, and the cost-effectiveness ratios were 21 709, 250 939, 76 870, 138 463 and 92 860 yuan (RMB)/couple, respectively. When the price of genetic testing was lower than 55 yuan (RMB), the cost-effectiveness ratio of genetic screening was lower than that of Mode 3.Conclusions:MCV<80 fl can be considered as the positive criteria in blood routine screening for thalassemia in Hunan Province, and the cost-effectiveness ratio of Mode 3 (the couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing) is the best. Genetic screening has certain advantages with the decreasing price.

Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×10⁴ U and 33.22×10⁴ U) was higher than that of BALB/c mice (5.33×10⁴ U and 19.31×10⁴ U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×10⁴ U/g) was higher than that of BALB/c mice (0.23×10⁴ U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.

AIM: To explore the effect of spinal anesthesia on ventricular arrhythmia and involved mechanisms in myocardial ischemia-reperfusion (MIR) rats. METHODS: The rat MIR model was made by occlusion the left anterior descending coronary artery for 30 minutes and reperfusion for 45 minutes. Bupivacaine (0.05 mL / 100 g, 1 mg / kg) was injected slowly via intrathecal for spinal anesthesia. The electromyelogram at T2 thoracic spinal cord was recorded. Ventricular arrhythmias, cardiac function, myocardial damage were assessed by electrocardiography, echocardiography and TTC or HE staining. RESULTS: MIR reduced left ventricular short-axis shortening (LVFS) and left ventricular ejection fraction (LVEF), caused myocardial histological damage and ventricular arrhythmias, promoted spinal electrical discharge frequency and amplitude in T2 dorsal horn. Spinal injection of bupivacaine could significantly reduce spinal cord electrical activities and eliminate MIR-induced arrhythmias. Moreover, bupivacaine also significantly improved MIR-induced myocardial histological damage and cardiac function inhibition. CONCLUSION: Spinal anesthesia can reduce ventricular arrhythmias induced by MIR. The mechanism may be related to the effect of abolishing spinal nerve excitability.

Background: Due to the unavailability of an effective vaccine or antiviral drug against the African swine fever virus (ASFV), rapid diagnosis methods are needed to prevent highly contagious African swine fever. Objectives: The objective of this study was to establish the ladder-shape melting temperature isothermal amplification (LMTIA) assay for the detection of ASFV. Methods: LMTIA primers were designed with the p72 gene of ASFV as the target, and plasmid pUC57 was used to clone the gene. The LMTIA reaction system was optimized with the plasmid as the positive control, and the performance of the LMTIA assay was compared with that of the commercial real-time polymerase chain reaction (PCR) kit in terms of sensitivity and detection rate using 200 serum samples. Results: Our results showed that the LMTIA assay could detect the 10 4 dilution of DNA extracted from the positive reference serum sample, which was the same as that of the commercial real-time PCR kit. The coincidence rate between the two assays was 100%. Conclusions The LMTIA assay had high sensitivity, good detection, and simple operation. Thus, it is suitable for facilitating preliminary and cost-effective surveillance for the prevention and control of ASFV.

Objective To investigate the expression of FNDC5 in hepatocellular carcinoma (HCC) and its relationship with clinicopathological characteristics and prognosis of HCC patients.Methods Immunohistochemistry and qRT-PCR were used to detect the expression of FNDC5 in HCC tissues,and the relationship between its expression and clinicopathological parameters and prognosis.Results FNDC5 was mainly expressed in the cytoplasm of HCC cells,weakly expressed in the nucleus.Among the 30 liver cancer tissues,FNDC5 was weakly positive in 5,positive in 19,and strongly positive in 6 cases.QRT-PCR assay showed that FNDC5 was highly expressed in HCC tissues with vascular invasion.The incidence of vascular invasion in the high-expression of FNDC5 was 30％ (9/30),which was significantly higher than that of the FNDC5 low-expression group [6.7％ (2/30),] and the difference was significant (x2 =15.026,P ＜0.05).There was no significant correlation between FNDC5 level and age,sex,HBsAg,and alpha-fetoprotein (AFP) level in HCC (P ＞ 0.05).Conclusion FNDC5 level in the liver cancer tissues is closely related to the occurrence of vascular invasion.

BACKGROUND: Mesenchymal stem cells can protect and repair the liver of rats with liver failure, but the mechanisms are not completely clear. OBJECTIVE: To explore the protective effects and related mechanisms of intravenous injection of human adipose-derived mesenchymal stem cells on acute liver failure in rats. METHODS: Thirty-six Sprague-Dawley rats (provided by Qingdao Daren Fucheng Animal Husbandry Co., Ltd. in China) were randomly divided into control group, model group and transplantation group. Animal models of acute liver failure were established by intraperitoneal injection of D-galactosamine in the model group and the transplantation group. One day after modeling, the rats in the transplantation group were injected with human adipose-derived mesenchymal stem cell suspension, and those in the model group were injected with the same amount of saline. After 1 and 3 days of cell transplantation, the serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were measured. Three days after cell transplantation, the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-10 were detected, the pathological changes of the rat liver were observed by hematoxylin-eosin staining, and the activity of glycogen synthase kinase-3β protein in the liver tissue was detected by western blot. RESULTS AND CONCLUSION: Compared with the model group, there was a significant reduction in the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, tumor necrosis factor-α, interleukin-6 and interleukin-10 in the transplantation group (P < 0.05). Inflammation and necrosis of liver tissues in the transplant group were alleviated compared with the model group. The activity of glycogen synthase kinase 3β in the liver tissue of the transplanted group was lower than that of the model group (P < 0.05). Overall, these results indicate that human adipose-derived mesenchymal stem cells can alleviate hepatic inflammation and pathological injury, and improve the liver function in rats with acute hepatic failure. Moreover, the mechanism may be related to the inhibition of glycogen synthase kinase 3β activity.

Objective: To investigate the expression and distribution of FNDC5/Irisin in pancreatic cancer tissues and adjacent tissues, and to analyze its correlation with clinicopathological features. Methods: Collection of archived wax blocks from 64 patients diagnosed with pancreatic cancer after surgical treatment from January 2015 to December 2018 in the Department of Pathology, Affiliated Qingdao Municipal Hospital of Qingdao University, and 30 tissues collected intraoperatively from January 2016 to December 2018 Samples, all collected samples included tumor tissue and corresponding adjacent tissues (>2 cm from the tumor edge). Real-time quantitative PCR (qRT-PCR) and immunohistochemistry were used to detect the expression of FNDC5/Irisin mRNA and its positivity in pancreatic cancer tissues and adjacent tissues, and to analyze its relationship with clinicopathological features of pancreatic cancer. Results: qRT-PCR showed that the expression of FNDC5/Irisin mRNA in pancreatic cancer tissues was higher than that in the corresponding adjacent pancreatic tissues, the difference was statistically significant (P<0.05). The immunohistochemistry results showed that the positivity of FNDC5/Irisin in pancreatic cancer tissues was 59.4%, and the positivity of FNDC5/Irisin in adjacent tissues was 28.1%, and the positivity of FNDC5/Irisin in cancer tissues and adjacent pancreatic tissues was significantly different (P<0.05); the expression level of FNDC5/Irisin was not related with the gender, age, tumor size, degree of differentiation, and tumor stage.(P>0.05), but FNDC5/Irisin expression was associated with liver and lymph node metastasis (P<0.05). Conclusion The positivity of FNDC5/Irisin in pancreatic cancer tissues is significantly higher than that in adjacent pancreatic tissues, and it is correlated with liver and lymph node metastasis in pancreatic cancer.

Objective To investigate the effect of Irisin on proliferation,apoptosis,migration and invasion of human cholangiocarcinoma cell line Hucct-1.Methods After treatment with Irisin,CCK-8 assay and flow cytometry assay were conducted to investigate the effect of Irisin on proliferation and apoptosis of cholangiocarcinoma cells.Scratch test and transwell invasion assay were used to studythe effect of Irisin on the migration and invasion ability of cholangiocarcinoma cells.Western blot was utilized to detect the expression of E-cadherin,N-cadherin and Vimentin in cholangiocarcinoma cells.Results CCK-8 assay showed that Irisin inhibited cholangiocarcinoma cell proliferationin a dose-dependent manner.Flow cytometry assay showed that the apoptosis rate of Irisin group [(14.8 ±0.9)％] was higher than that in the control group [(5.4±0.6)％],(P＜0.05).The scratch test showed that the rate of cell scratch healing in Irisin group [(15.0± 1.0)％] was significantly lower than that in the control group [(28.0±2.0)％] (P＜0.05).Transwell invasion test showed that the number of cells in Irisin group was (96.0±7.0),which was significantly lower than that in control group (155.0± 9.0) (P＜0.05).Western blot showed that the expression of E-cadherin increased and N-cadherin and Vimentin decreased after Irisin treatment.Conclusion Irisin inhibits proliferation,migration and invasion and promote apoptosis of cholangiocarcinoma cells.

Sinus floor elevation was needed in 11 patients having 15 implant sites with the residual bone height (RBH) was less than 10 mm in the posterior maxillary region from Feb to May 2017. The RBH ranged from 3.10 to 8.34 mm [averaged (6.18±1.60) mm]. RBH<6 mm was observed in 40% implant sites (6 implant sites) and RBH≥6 mm was observed in 60% implant sites (9 implant sites). The thickness of the sinus floor membrane correspond to the implant site measured by cone beam CT (CBCT) ranged from 0.50 to 4.24 mm [averaged (1.21±0.92) mm]. Sequential drills with stops were used to perforate the cortical bone of the sinus floor firstly, then the transcrestal around detached sinus floor elevation technique (TADSFET) was carried with osteotomes. Anorganic bovine bone was used as the augmentation material.Fifteen implants were placed in 15 implant sites. CBCT pictures showed that there was a smooth and continuous tent-shaped apophysis on each lifted site and no air fluid level was observed in the sinus immediately after operation. The mean elevated height of the 15 implant sites was (7.83±1.57) mm (ranged from 5.94 to 11.01 mm). The mean follow-up time was 7.91 months (7-10 months). The survival rate was 100% during the follow up period.