Clinical effectiveness of sodium fluorescein-guided microsurgery in patients with high-grade gliomas.

OBJECTIVE: The aim of this study was to assess the clinical effectiveness of sodium fluorescein-guided microsurgery in patients with high-grade gliomas. PATIENTS AND METHODS: 120 patients with high-grade gliomas who were hospitalized in our Neurosurgery Department from January 2018 to January 2021 were selected and then divided into a control and a study group using the random number table method, with 60 cases in each group. To compare the clinical efficacy of patients in both groups, neuronavigation microsurgery was used in the control group and neuronavigation microsurgery combined with sodium fluorescein-guided microsurgery was used in the study group. RESULTS: The Gross Total Resection Rate (GTRR) of the study group was significantly higher than that of the control group. There was no significant difference in intraoperative bleeding loss or hospital stay between the two groups, and the study group had a much shorter operation time than the control group. The Karnofsky Performance Score (KPS) and the National Institutes of Health Stroke Scale (NIHSS) scores did not significantly differ between the two groups prior to surgery but declined significantly in the study group compared to the control group following treatment. In terms of adverse effects, there was no significant difference between the two groups. In the control group, the median progression-free survival (PFS) was 7.5 months, and the median overall survival (OS) was 9.6 months, whereas in the study group, the median PFS was 9.5 months, and the median OS was 11.5 months. PFS did not significantly differ between the two groups (HR=1.389, 95% CI=0.926-2.085, p=0.079); however, OS was significantly higher in the study group compared to the control group (HR=1.758, 95% CI=1.119-2.762, p=0.013). CONCLUSIONS: Fluorescein-guided microsurgery can dramatically improve total resection rate, postoperative neurological functional status, and overall survival with higher efficacy and safety in patients with high-grade gliomas.

[Safety and efficacy of laparoscopic surgery in locally advanced gastric cancer patients with neoadjuvant chemotherapy combined with immunotherapy].

Objective: To investigate the safety and efficacy of laparoscopic surgery in locally advanced gastric cancer patients with neoadjuvant SOX chemotherapy combined with PD-1 inhibitor immunotherapy. Methods: Between November 2020 and April 2021, patients with locally advanced gastric cancer who were admitted to the Union Hospital of Tongji Medical College of Huazhong University of Science and Technology were prospectively enrolled in this study. Inclusion criteria were: (1) patients who signed the informed consent form voluntarily before participating in the study; (2) age ranging from 18 to 75 years; (3) patients staged preoperatively as cT3-4N+M0 by the TNM staging system; (4) Eastern Collaborative Oncology Group score of 0-1; (5) estimated survival of more than 6 months, with the possibility of performing R0 resection for curative purposes; (6) sufficient organ and bone marrow function within 7 days before enrollment; and (7) complete gastric D2 radical surgery. Exclusion criteria were: (1) history of anti-PD-1 or PD-L1 antibody therapy and chemotherapy; (2) treatment with corticosteroids or other immunosuppre- ssants within 14 days before enrollment; (3) active period of autoimmune disease or interstitial pneumonia; (4) history of other malignant tumors; (5) surgery performed within 28 days before enrollment; and (6) allergy to the drug ingredients of the study. Follow-up was conducted by outpatient and telephone methods. During preoperative SOX chemotherapy combined with PD-1 inhibitor immunotherapy, follow-up was conducted every 3 weeks to understand the occurrence of adverse reactions of the patients; follow-up was conducted once after 1 month of surgical treatment to understand the adverse reactions and survival of patients. Observation indicators were: (1) condition of enrolled patients; (2) reassessment after preoperative therapy and operation received (3) postoperative conditions and pathological results. Evaluation criteria were: (1) tumor staged according to the 8th edition of the American Joint Committee on Cancer (AJCC) TNM staging system; (2) tumor regression grading (TRG) of pathological results were evaluated with reference to AJCC standards; (3) treatment-related adverse reactions were evaluated according to version 5.0 of the Common Terminology Criteria for Adverse Events; (4) tumor response was evaluated by CT before and after treatment with RECIST V1.1 criteria; and (5) Clavien-Dindo complication grading system was used for postoperative complications assessment. Results: A total of 30 eligible patients were included. There were 25 males and 5 females with a median age of 60.5 (35-74) years. The primary tumor was located in the gastroesophageal junction in 12 cases, in the upper stomach in 8, in the middle stomach in 7, and in the lower stomach in 3. The preoperative clinical stage of 30 cases was III. Twenty-one patients experienced adverse reactions during neoadjuvant chemotherapy combined with immunotherapy, including four cases of CTCAE grade 3-4 adverse reactions resulting in bone marrow suppression and thoracic aortic thrombosis. All cases of adverse reactions were alleviated or disappeared after active symptomatic treatment. Among the 30 patients who underwent surgery, the time from chemotherapy combined with immunotherapy to surgery was 28 (23-49) days. All 30 patients underwent laparoscopic radical gastrectomy, of which 20 patients underwent laparoscopic-assisted radical gastric cancer resection; 10 patients underwent total gastrectomy for gastric cancer, combined with splenectomy in 1 case and cholecystectomy in 1 case. The surgery time was (239.9±67.0) min, intraoperative blood loss was 84 (10-400) ml, and the length of the incision was 7 (3-12) cm. The degree of adenocarcinoma was poorly differentiated in 18 cases, moderately differentiated in 12 cases, nerve invasion in 11 cases, and vascular invasion in 6 cases. The number lymph nodes that underwent dissection was 30 (17-58). The first of gas passage, the first postoperative defecation time, the postoperative liquid diet time, and the postoperative hospitalization time of 30 patients was 3 (2-6) d, 3 (2-13) d, 5 (3-12) d, and 10 (7-27) d, respectively. Postoperative complications occurred in 23 of 30 patients, including 7 cases of complications of Clavien-Dindo grade IIIa or above. Six patients improved after treatment and were discharged from hospital, while 1 patient died 27 days after surgery due to granulocyte deficiency, anemia, bilateral lung infection, and respiratory distress syndrome. The remaining 29 patients had no surgery-related morbidity or mortality within 30 days of discharge. Postoperative pathological examination showed TRG grades 0, 1, 2, and 3 in 8, 9, 4, and 9 cases, respectively, and the number of postoperative pathological TNM stages 0, I, II, and III was 8, 7, 8, and 7 cases, respectively. The pCR rate was 25.0% (8/32). Conclusion: Laparoscopic surgery after neoadjuvant SOX chemotherapy combined with PD-1 inhibitor immunotherapy for locally advanced gastric cancer is safe and feasible, with satisfactory short-term efficacy. Early detection and timely treatment of related complications are important.

[Techniques in prophylactic ileostomy reversal].

In order to prevent and reduce the severity of anastomotic leakage after low rectal cancer surgery, prophylactic ileostomy is often performed by the clinician simultaneously. There are many controversies about prophylactic ileostomy in medicine, such as ileostomy indications, ileostomy complications, ileostomy reversal time, ileostomy reversal method and technique. Based on relevant literature and our own experience, we discussed the timing, method and complications of ileostomy reversal in this article to improve the diagnosis and treatment of ileostomy reversal as well as the life quality of the patients after ileostomy reversal.

[Clinicopathological characteristics and prognosis of patients with sporadic multiple primary gastrointestinal stromal tumors].

Objective: To investigate the clinicopathological characteristics and prognosis of sporadic multiple primary gastrointestinal stromal tumor (GIST). Methods: A retrospective cohort study was conducted. Case inclusion criteria: (1) postoperative pathological diagnosis of GIST; (2) primary GIST with single lesion or sporadic multiple primary GIST (sporadic GIST was defined as primary GIST other than familial and syndrome-related GIST, and multiple primary GIST was defined as the number of primary GISTs in the same patient ≥ 2); (3) patients with complete clinicopathological data. Those with tumor recurrence or distant metastasis, and with other malignancies were excluded. Medical records of patients with primary GIST who underwent surgical resection in the Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2010 to December 2020 were collected. Patients were divided into sporadic multiple primary GIST group and single primary GIST group according to the number of primary GIST lesions. The clinicopathological data and prognosis of the two groups were observed and compared. Results: A total of 1200 patients with primary GIST were enrolled in this study, including 628 males (52.3%) and 572 females (47.7%), with a median onset age of 58 (19-93) years. Among them, 1165 cases (97.1%) were sporadic primary GIST with single lesion; 35 cases (2.9%) were sporadic multiple primary GIST. Among 35 cases of sporadic multiple primary GIST, 3 cases (8.6%) had acid reflux as the first symptom, which was higher than the single primary GIST group (22/1165, 1.9%) (χ(2)=7.437, P=0.006). There were no significant differences in other clinical characteristics between the two groups (all P>0.05). Patients in the sporadic multiple primary GIST group contained a total of 80 primary tumors. Compared with the single primary GIST group, the sporadic multiple primary GIST group had a higher proportion of tumors originating in the stomach [87.5% (70/80) vs. 59.1% (689/1165)], lower proportion of spindle cell in histology [85.0% (68/80) vs. 93.7% (1092/1165)], higher proportion of positive CD34 [97.5% (78/80) vs. 87.6% (1021/1165)], smaller maximum diameter [maximum diameter ≤2.0 cm: 61.2% (49/80) vs. 28.8% (335/1165)], lower mitotic rate [≤5/50 high-power fields (HPF): 93.8% (75/80) vs. 74.5% (868/1165)], lower risk of recurrence [60.0% (48/80) vs. 23.3% (271/1165)], and the differences were all statistically significant (all P<0.05). The 3-year recurrence-free survival rate in the sporadic multiple primary group and the single primary GIST group was 96.6% and 89.3% respectively (P=0.160), and the 3-year overall survival rate was 100.0% and 92.8%, respectively (P=0.088). Conclusions: The most common type of sporadic multiple primary GIST is multiple tumors originating in the stomach at the same time. Compared with primary GIST with single lesion, sporadic multiple primary GIST presents smaller maximum diameter and lower mitotic rate. The prognosis of patients between two groups is not significantly different.

[Application of three-dimensional digital simulated design and implementation in esthetic rehabilitation].

Objective: To explore and evaluate a three-dimensional (3D) digital simulated design and implementation technique in esthetic rehabilitation. Methods: Thirty patients with esthetic deficiency, who came to the Department of Prosthodontics, Peking University School and Hospital of Stomatology from December 2017 to July 2019, were recruited and randomly assigned into 2 groups. Four males and 11 females which were (36.0±10.5) years old in the experimental group, 6 males and 9 females which were (32.0±6.7) years old in the control group, were enrolled. In the experimental group, 3D digital simulated design was used to predict the post-treatment effect; and the final restorations were designed duplicating from the pre-treatment digital design confirmed by the patient and milled. In the control group, the final restorations were manufactured by the dental technician according to the design of two-dimensional (2D) digital smile design. The simulation degree of digital design and the similarity between preoperative design and postoperative rehabilitation were scored by the patients with visual analogue scales (VAS) in both groups; and the satisfaction rate to the restorations was evaluated by the patients. The quality of the restorations was accessed by a prosthodontist who did not know the grouping of patients according to the modified criteria of United States Public Health Service (USPHS). Results: Three-dimensional digital simulated design and implementation technique was successfully established. The VAS score on the simulation degree of digital design in the experimental group (8.5±0.5) was higher than that in the control group (7.2±0.7) (P<0.01); the VAS score on the similarity between preoperative design and postoperative rehabilitation in the experimental group (9.6±0.3) was higher than that in the control group (7.0±0.9) (P<0.01). The satisfaction rate of the patients to the restorations was significantly higher in the experimental group than in the control group (P<0.05). There was no significant difference of the quality of the restorations between the two groups on the anatomic form, the marginal adaption and the surface quality (P>0.05). Conclusions: Three-dimensional digital simulated design and implementation technique can help achieving 3D digital simulated design before treatment and duplication to the final restorations, and can improve the patients' satisfaction in esthetic rehabilitation.

[Recommendations for general surgery clinical practice in 2019 coronavirus disease situation].

The 2019 coronavirus disease(COVID-19) is a highly infectious disease, has a long incubation period and a variety of clinical manifestations, which has a significant impact on public health and life. Afterwards, scientific and standardized work processing during the epidemic is of great significance for prevention and control. In order to implement the central government's decision-making deployment and defeat the COVID-19 as soon as possible, we had focused on the key points in the clinical work of general surgery according to latest relevant guidelines, literature and experience in epidemic prevention. Finally, we drafted the prevention and control strategies and recommendations to make a reference for medical staff of general surgery to fight against COVID-19.

[Recommendations for general surgery clinical practice in novel coronavirus pneumonia situation].

Novel coronavirus pneumonia (NCP) is a highly infectious disease, has a long incubation period and a variety of clinical manifestations, which has a significant impact on public health and life. Afterwards, scientific and standardized work processing during the epidemic is of great significance for prevention and control. In order to implement the central government's decision-making deployment and defeat the NCP as soon as possible, we had focused on the key points in the clinical work of general surgery according to latest relevant guidelines, literature and experience in epidemic prevention. Finally, we drafted the prevention and control strategies and recommendations to make a reference for medical staff of general surgery to fight NCP.

[Early postoperative complications and risk factors in laparoscopic D2 radical gastrectomy for gastric cancer].

Objective: To investigate the morbidity and treatment of early postoperative complications after laparoscopic D2 radical gastrectomy for gastric cancer, and to explore the risk factors. Methods: A case-control study was performed to retrospectively collect clinicopathological data of 764 patients undergoing laparoscopic D2 radical gastrectomy for gastric cancer at our department between January 2015 and December 2017. Patient inclusion criteria: (1) gastric cancer diagnosed by preoperative electronic gastroscopy and biopsy, and confirmed by postoperative pathology; (2) without invasion into adjacent organs by preoperative evaluation of tumors; (3) tumors without definite liver and distant metastasis; (4) R0 resection of gastric cancer and standard D2 lymph node dissection; (5) patients with informed consent. Exclusion criteria: (1) unperformed laparoscopic D2 radical resection; (2) other types of gastric tumor confirmed by pathology; (3) cases with incomplete clinical data. Complication occurring within two weeks after laparoscopic D2 gastrectomy was defined as early postoperative complication. Patients were divided into two groups: non-complication group (693 cases) and complication group (71 cases) according to the occurrence of complications after operation. The clinicopathological data of two groups were analyzed and compared with t test and χ(2) test, and the factors of P < 0.2 were included in the multivariate logistic regression model to analyze the risk factors of postoperative complications. Results: Of 764 patients, 71 (9.3%) developed early postoperative complications, with median onset time of 3 (1 to 11) days. Surgical complications accounted for 7.9% (60/764), including 13 cases (1.7%) of abdominal hemorrhage, 12 cases (1.6%) of anastomotic leakage, 10 cases (1.3%) of incision infection, 8 cases (1.0%) of anastomotic bleeding, 7 cases (0.9%) of gastric stump weakness, 4 cases (0.5%) of abdominal infection, 4 cases (0.5%) of duodenal stump leakage and 2 cases (0.3%) of small intestinal obstruction. Non-surgical complications accounted for 1.4% (11/764), including 6 cases (0.8%) of pulmonary infection and 5 cases (0.7%) of cardiovascular disease. Two cases (0.3%) died of sepsis caused by severe abdominal infection; 9 cases (1.2%) recovered after receiving the second operation, among whom 5 cases were abdominal hemorrhage, 2 cases were anastomotic leakage and 2 cases were duodenal stump leakage; the remaining patients were healed with conservative treatment. Compared with patients without complications, patients with complications had higher proportions of BMI ≥24 kg/m(2) [42.3% (30/71) vs. 24.2%(168/693), χ(2)=10.881, P=0.001], comorbity [64.8% (46/71) vs. 33.5% (232/693), χ(2)=27.277, P<0.001], combined organ resection [70.4% (50/71) vs. 20.5% (142/693), χ(2)=85.338, P<0.001], and pTNM stage of III [70.4% (50/71) vs. 40.1% (278/693), χ(2)=24.196, P<0.001], meanwhile had longer time to postoperative flatus [(4.2±2.1) days vs. (2.9±1.2) days, t=4.621, P=0.023], longer hospital stay [(34.6±12.6) days vs. (14.2±6.2) days, t=9.862, P<0.001] and higher hospitalization cost [(126.8±64.5) thousand yuan vs. (85.2±35.8) thousand yuan, t=11.235, P<0.001]. Multivariate analysis showed that BMI ≥24 kg/m(2) (OR=3.762, 95% CI: 1.960-8.783, P=0.035), accompanying disease (OR=8.620, 95% CI: 1.862-29.752, P<0.001), combined organ resection (OR=6.210, 95% CI: 1.357-21.568, P=0.026), and pTNM stage (OR=4.752, 95% CI: 1.214-12.658, P<0.001) were the independent risk factors of postoperative complications. Conclusions: Laparoscopic D2 radical gastrectomy is a safe and effective approach for gastric cancer. Most early postoperative complications can obtain satisfactory efficacy after conservative treatment. Perioperative management should be strengthened for those patients with high BMI, accompanying diseases, combined organ resection, and advanced pTNM stage.

[Laparoscopic versus open surgery for gastric gastrointestinal stromal tumors in unfavorable location: a propensity score-matching analysis].

Objective: To investigate the efficacy and feasibility of laparoscopic resection for gastric gastrointestinal stromal tumor (GIST) in unfavorable location by comparing with open surgery. Methods: Clinicopathological and follow-up data of 176 patients with gastric GIST in unfavorable location admitted at Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2005 to December 2017 were analyzed retrospectively. There were 94 males and 82 females, aging of (57.4±12.7) years (range: 20-90 years). Of the 176 patients, 64 underwent laparoscopic surgery (laparoscopic group) and 112 underwent open surgery (open group). One-to-one propensity score matching (PSM) was performed to balance the covariance between laparoscopic group and open surgery group. Before PSM, the differences between the two group in tumor size and modified National Institutes of Health risk classification were significant. After PSM, there were 63 pairs (63 cases in laparoscopic group and 63 cases in open group) and the baseline characteristics were comparable between the two groups(P>0.05). The difference of short-term outcome between the two groups were compared using t test, χ(2) test or Wilcoxon rank-sum test. The survival curve was established by Kaplan-Meier method and the Log-rank test was used to compare the survival of the two groups. Results: The operation time of laparoscopic group was shorter ((141.6±100.6) minutes vs. (100.4±67.7) minutes, t=2.681, P=0.008), the hospitalization cost was higher ((5.2±0.7) ten thousand yuan vs. (4.2±0.8) ten thousand yuan, t=7.357, P=0.000) than open group. The time to first flatus ((49.1±8.2) hours vs. (71.0±4.6) hours, t=-18.482, P=0.000) and preoperative hospital stay ((10.3±6.0) days vs. (14.8±7.6) days, t=-3.717, P=0.000) was shorter in laparoscopic group. With a median follow-up time of 44 months (range: 10 to 154 months), the 1-, 3-, 5-year relapse-free survival rates in the laparoscopic group and open group were 98.3%, 92.1%, 92.1% and 100%, 86.3%, 83.2%, respectively (χ(2)=0.696, P=0.404). The 1-, 3-, 5-year overall survival rates in the laparoscopic group and open group were 96.6%, 94.7%, 94.7% and 100%, 91.1%, 81.4%, respectively (χ(2)=0.366, P=0.545). Conclusions: In experienced medical centers, laparoscopic resection is safe and feasible for GIST in unfavorable location. Compared to open surgery, laparoscopic resection achieves a faster postoperative recovery and a similar long-term prognosis.

[Preliminary clinical application of complete digital workflow of design and manufacturing occlusal splint for sleep bruxism].

OBJECTIVE: To establish a complete workflow of digital design and manufacturing occlusal splint for sleep bruxism, which can be preliminarily applied in clinical use, thus observe the clinical efficacy. METHODS: Twenty-four patients with sleep bruxism were recruited in the study and randomly divided into two groups by using random number tables. Digital-occlusal-splint (experimental group) treatment plan and traditional-occlusal-splint (control-group) treatment plan were carried out for each group, respectively. For experimental group, digital models of patients' both dental arches and the occlusion relationship after elevation were captured using an intraoral scanner. The occlusal splint was carried out by computer aided design/computer aided manufacturing (CAD/CAM), including splint designing and milling. For control group, the traditional soft occlusal splint was fabricated by vacuum laminator. The two kinds of occlusal splints were tried in the patients from each group, and the occlusal contacts were tested respectively by T-scan analysis system, which recorded the changes of occlusal indicators in the two groups. The retention, appearance and occlusal comfort degree were evaluated by the two groups of patients. Mann-Whitney test was performed with IBM SPSS 20.0 software, and bilateral test was performed. P<0.05 was considered to be statistically significant. RESULTS: The complete workflow of digital design and manufacturing occlusal splint was successfully established. During the clinical use, there was no statistical difference in the retention evaluation of two kinds of occlusal splints between the two groups of patients (Z=-0.538, P=0.590). The appearance score (Z=2.038, P=0.042) and the occlusal comfort score (Z=-2.579, P=0.010) of the experimental group were higher than those of the control group, with statistically significant differences. The T-scan analysis results showed that only the second molar on both sides of the traditional occlusal splint had occlusal contact in intercupsal position, while the digital occlusal splint had stable and bilaterally balanced contact between the maxillary and mandibular teeth. Furthermore, the occlusal force was uniformly distributed in the experimental group. CONCLUSION: The complete workflow of digital occlusal splint improves the occlusal design, greatly simplifies and optimizes the traditional process of making occlusal splint. This new method is resource-saving and environmental-friendly, and it is able to serve patients more conveniently and efficiently.

Using magnetic resonance diffusion tensor imaging to evaluate renal function changes in diabetic patients with early-stage chronic kidney disease.

AIM: To investigate the clinical value of diffusion tensor imaging (DTI) in assessing renal function changes in diabetic patients with early-stage chronic kidney disease (CKD), and the relationship of DTI parameters with estimated glomerular filtration rate (eGFR) and urinary biomarkers. MATERIALS AND METHODS: Thirty-six patients with diabetes mellitus (DM; 30 CKD stage 1 and 6 CKD stage 2) and 26 healthy control subjects were enrolled. DTI was performed using a clinical 3 T MRI system. Apparent diffusion coefficient (ADC) and fractional anisotropy (FA) values were calculated from the renal cortex and medulla. The correlation of the DTI parameters with eGFR and urinary biomarkers was evaluated. RESULTS: FA values were significantly reduced in the renal cortex and medulla of DM group compared with the control group (cortical FA, Z=-2.834, p=0.005; medullary FA, t=2.768, p=0.007). In the DM group, FA values in the renal cortex and medulla were positively correlated with eGFR, while FA values in the medulla were negatively correlated with the urinary albumin/creatinine ratio, urinary alpha-1 microglobulin/creatinine ratio, and urinary transferring/creatinine ratio. ADC values in the renal cortex and medulla showed a trend towards an increase in the DM group compared with the control group. CONCLUSIONS: Renal DTI is a promising method for assessing early renal function changes in DM patients.

Effect of mannose on the lung function of rats with acute pancreatitis.

The present study aimed to investigate the mechanisms by which mannose protects the lung injury induced in rats with acute pancreatitis (AP). An AP combined with Acute Lung Injury (ALI) model was established. A total of 90 healthy adult male Sprague-Dawley rats (300±50g weight) were randomly divided into three groups: sham operation group (SO group), severe acute pancreatitis lung injury group (SAP group), and mannose intervention group (MT group). Subsequently, each group was divided into two subgroups based on the time passed from intervention, namely 6 and 12 h. Each subgroup comprised 15 rats. The ratio of wet/dry weight of the lung tissue exhibited no significant change at different time points in the SO group. This parameter was significantly increased in the SAP group compared with the SO group at each time point of the treatment (P less than 0.05) and it was significantly lower in the MT group than that in the AP group (P less than 0.05) and it was significantly increased in the AP group at each time (P less than 0.05) compared with the SO group. The levels of TNF-α in the lung tissue in the SO group exhibited no significant change at different time points, but they were significantly decreased in the MT group at each time point (P less than 0.05) compared with the SAP group. The mannose receptor (MR) mRNA and protein levels in the lung tissues exhibited no significant change at different time points. The mRNA and protein levels of MR in the SAP group were significantly decreased at each time point (P less than 0.05) compared with the SO group. The mRNA and protein levels of MR, in the lung tissue of the MT group were significantly increased at each time point compared with the SAP group (P less than 0.05). Mannose could reduce the injury caused to the lung tissue of rats with severe acute pancreatitis by up-regulation of the expression of MR mRNA and protein.

Seroprevalence of Toxoplasma gondii infection in sheep from northern China.

Toxoplasma gondii is an important zoonotic parasite causing significant health problems to humans and animals. In recent years, a number of investigations about the seroprevalence of T. gondii in China have been reported, but little is known on the prevalence of toxoplasmosis in sheep in northern China. In the present study, a total of 288 sheep serum samples were collected from Inner Mongolia, Heilongjiang, Jilin and Hebei provinces of northern China for T. gondii antibody survey using a latex agglutination test (LAT). Of these, 87 (30.2%) serum samples were positive for antibodies to T. gondii, and the antibody titres ranged from 1:64 to 1:1,024. Seroprevalence of T. gondii infection in sheep was 17.1% in Inner Mongolia, 33.8% in Heilongjiang, 24.6% in Jilin and 46.3% in Hebei. Age and rearing system significantly affected seropositivity. The present survey indicates antibodies to T. gondii are widely prevalent in sheep in northern China, which may cause public health problems in these provinces.

Seroprevalence of Toxoplasma gondii infection in sheep from northern China

@#Toxoplasma gondii is an important zoonotic parasite causing significant health problems to humans and animals. In recent years, a number of investigations about the seroprevalence of T. gondii in China have been reported, but little is known on the prevalence of toxoplasmosis in sheep in northern China. In the present study, a total of 288 sheep serum samples were collected from Inner Mongolia, Heilongjiang, Jilin and Hebei provinces of northern China for T. gondii antibody survey using a latex agglutination test (LAT). Of these, 87 (30.2%) serum samples were positive for antibodies to T. gondii, and the antibody titres ranged from 1:64 to 1:1,024. Seroprevalence of T. gondii infection in sheep was 17.1% in Inner Mongolia, 33.8% in Heilongjiang, 24.6% in Jilin and 46.3% in Hebei. Age and rearing system significantly affected seropositivity. The present survey indicates antibodies to T. gondii are widely prevalent in sheep in northern China, which may cause public health problems in these provinces.

MiR-21 enhanced glioma cells resistance to carmustine via decreasing Spry2 expression.

OBJECTIVE: Gliomas are accompanied with high mortality owning to their invasive peculiarity and vulnerability to drug resistance. miR-21 is a vital oncogenic miRNA that regulates drug resistance of tumor cells. This study aims to elucidate the function of miR-21 in human glioma cells resistant to carmustine (BCNU) and to demonstrate the underlying molecular mechanism. MATERIALS AND METHODS: BCNU-sensitive cells (SWOZ2 cells) were transfected with miR-21 agomir and negative control, and BCNU-resistance cells (SWOZ2-BCNU cells) were transfected with miR-21 antagomir and negative control. The Real-time fluorescence quantitative PCR was used to detect and compare the levels of miR-21expression between SWOZ2-BCNU and SWOZ2 cells. The drug sensitivity of these cells to BCNU was determined by Cell Counting Kit-8 (CCK-8) assay. The protein expression of Spry2 was detected by Western blotting. RESULTS: The expression level of miR-21 was remarkably higher in SWOZ2-BCNU cells than that in SWOZ2 cells. The half-maximal inhibitory concentration (IC50) of BCNU was obviously higher for SWOZ2-BCNU cells than that for SWOZ2 cells. Besides, we found that aberrant expression of miR-21 in SWOZ2-BCNU cells is responsible for glioma BCNU-resistance. Consistently, Spry2 protein levels were significantly reduced in SWOZ2-BCNU as well as in miR-21 agomir-transfected cells, inversely correlated to miR-21 expression. The results of si-Spry2 co-transfection suggested that the effect of miR-21 on glioma BCNU-resistance is mediated through Spry2. CONCLUSIONS: miR-21 enhances the resistance of human glioma cells to BCNU by decreasing the expression of Spry2 protein. Thus, Spry2 may be a novel therapeutic target for treating glioma BCNU-resistance.

Mechanism of As2O3 induces apoptosis of glioma U87 cells.

OBJECTIVE: To investigate the mechanism of arsenic trioxide (As2O3)-induced apoptosis of glioma cells. MATERIALS AND METHODS: U87 cells were treated by different concentrations of As2O3 (8 µmol/L, 6 µmol/L, 4 µmol/L, 2 µmol/L, 1 µmol/L and 0.5 µmol/L) for 24 h, 48 h and 72 h, respectively. Cell viability was detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the appropriate dosage and time were screened. Transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) was used to stain cells, followed by an investigation on the apoptosis of cells. In the study of molecular mechanism, the expression of p53 in the cells was determined by immunofluorescence, and then apoptosis-related factors, Fas, FasL and Bax, were tested using Real-time polymerase chain reaction (RT-PCR). Finally, the effect of As2O3 on apoptosis-related proteins, caspase-3 and caspase-9, was investigated by Western blotting. RESULTS: As2O3 could significantly inhibit proliferation of U87 cells, and the result of TUNEL staining displayed As2O3 had the function of inducing apoptosis. Immunofluorescence results demonstrated that p53 was highly expressed in glioma cells, which was reduced after drug administration. The results of detection of apoptosis factors using RT-PCR revealed that mRNA expressions of Fas, FasL and Bax in the glioma cells were distinctly higher than those in the As2O3 group. The result of Western blotting indicated that caspase-3 and caspase-9 proteins were highly expressed in glioma cells. Analysis of variance showed that the difference between the control group and the As2O3 group was statistically significant (p<0.01). CONCLUSIONS: As2O3 can inhibit proliferation of glioma cells and induce its apoptosis, which may be correlated with down-regulation of expressions of apoptosis-related factors, Fas, FasL and Bax, and apoptosis-related proteins, p53, caspase-3 and caspase-9.

[Role of ash2 (absent, small, or homeotic)-like and Jumonji domain-containing protein 3 on histone methylation of interferon-gamma gene and their associations with vascular damage of Kawasaki disease].

Objective: To investigate the impacts of ash2 (absent, small, or homeotic)-like (Ash2L) and Jumonji domain-containing protein 3 (Jmjd3) on histone methylation of interferon-gamma(IFN-γ) gene and association with vascular damage of Kawasaki disease (KD) in acute phase. Methods: This study was performed among 36 children with KD in acute phase (KD group) and 28 age-matched health children (control group), who were treated or underwent physical examination in our hospital between February 2015 and June 2016. Patients were further divided into KD groups with or without coronary artery lesions (KD-CAL(+) , 16 cases; KD-CAL(-), 20 cases). All KD patients were treated with intravenous immunoglobulin. The proportion of type 1 helper T(Th1) cells and protein levels of IFN-γ, T-box expressed in T cells(T-bet), phosphorylated signal transducer and activator of transcription 1(pSTAT1) and phosphorylated signal transducer and activator of transcription 4(pSTAT4) were analyzed by flow cytometry.Chromatin immunoprecipitation was performed to determine histone methylation (histone H3 tri-methyl K4(H3K4me3), histone H3 tri-methyl K27(H3K27me3)) and binding levels of Ash2L, Jmjd3 and Ezh2 associated with IFN-γ in CD4(+) T cells. Quantitative real-time PCR was used to determine mRNA levels of IFN-γ, interferon γ receptor 1(IFN-γR1), interferon γ receptor 2(IFN-γR2), interleukin 12 receptor subunit beta 1(IL-12Rß1), interleukin 12 receptor subunit beta 2(IL-12Rß2), interleukin 18 receptor subunit beta α(IL-18Rα), interleukin 18 receptor subunit beta ß(IL-18Rß), tumor necrosis factor receptor 1(TNFR1), toll-like receptor 4(TLR4), receptor interacting serine/threonine kinase 1(RIP-1) and myeloid differentiation primary response gene 88(MyD88) in CD4(+) T cells. Plasma concentrations of IFN-γ, interleukin 12(IL-12), interleukin 18(IL-18) and tumor necrosis factor α(TNF-α) were measured by enzyme-linked Immunosorbent assay. Results: (1)The proportion of Th1 and its protein level of IFN-γ were significantly higher in KD group than those in control group and higher in KD-CAL(+) group than in KD-CAL(-) group (all P<0.05), and lower after treatment than before treatment (all P<0.05). (2)Compared with control group, mRNA level of IFN-γ and IFN-γ-associating H3K4me3 was increased, while level of IFN-γ associating H3K27me3 in CD4(+) T cells was reduced in KD group (all P<0.05), which resulted in a higher rate of H3K4me3/H3K27me3 (P<0.05) in KD group, which was positively correlated with IFN-γ mRNA in KD group(r=0.55, P<0.05). Similar results were found between KD-CAL(+) group and KD-CAL(-) group (all P<0.05). Level of IFN-γ associating H3K27me3 was increased, and mRNA level of IFN-γ and IFN-γ associating H3K4me3 was decreased after treatment than before treatment (all P<0.05). (3)Expression of T-bet protein and binding levels of Ash2L and Jmjd3 with IFN-γ gene were significantly higher in KD group than those in control group(all P<0.05), higher in KD-CAL(+) group than those in KD-CAL(-) group (all P<0.05). These parameters were significantly lower after treatment than before treatment (all P<0.05). Binding level of Ezh2 with IFN-γ gene was similar among various groups (all P>0.05). (4)In comparison with control or after treatment, surface receptors(IFN-γR1/2, IL-12Rß1/2, IL-18Rα/ß, TNFR1 and TLR4) and its downstream molecules(pSTAT1, pSTAT4, RIP(1) and MyD88) in CD4(+) T cells, and plasma concentrations of inflammatory cytokines(IFN-γ, IL-12, IL-18 and TNF-α) were found to be higher in KD group(all P<0.05). These parameters were also higher in KD-CAL(+) group than in KD-CAL(-) group (all P<0.05). Conclusion: Aberrant histone methylation of IFN-γ associating H3K4me3 and H3K27me3 caused by over-binding of Ash2L and Jmjd3 might be involved in immune dysfunction and vascular damage in KD in the acute phase.

Expression and clinical significance of rhubarb on serum amylase and TNF-alpha of rat model of acute pancreatitis.

The aim of this study was to evaluate the therapeutic effect of rhubarb extract on acute pancreatitis. Ninety-six healthy Sprague Dawley rats, weighing 301±5.12 g were randomly divided into 4 groups: sham surgery (group A), acute pancreatitis model (group B), acute pancreatitis with normal saline (group C), and acute pancreatitis model with rhubarb (group D). The levels of serum amylase (AMY) and TNF-α were measured at 1st, 6th, 12th and 24th hour after modeling, and the pancreatic tissue were used to observe the pathologic changes. Compared to the sham group, the serum AMY and serum tumor necrosis factor (TNF-α) levels were significantly increased in the other groups (p <0.05). Compared to the model group and the saline group, the serum AMY, serum TNF-α level and pathological changes of rats in the rhubarb group were significantly lower (p <0.05). The serum AMY and TNF-α levels increased in acute pancreatitis. The rhubarb reduced the serum AMY and TNF-α level in rats with acute pancreatitis and reduced the pathological changes of pancreas and other tissues.

Molecular Detection of Theileria species in Cattle from Jilin Province, China.

Bovine theileriosis is a tick-borne disease that is hampering the development of the domestic cattle industry in northern China. This study involved a molecular survey of bovine Theileria species in 137 blood samples from cattle in the Jilin province of China. The DNA samples were screened by species-specific 18S rRNA PCR. Results revealed that 19.7% (27/137), 17.5% (24/137) and 10.9% (15/137) were found to be infected with Theileria sinensis, Theileria orientalis, respectively. Mixed infection was found in 8.8% (12/137). The overall detection rates of Baishan, Yanji, Jilin and Liaoyuan districts was 60.0%, 17.5%, 5.3% and 0%, respectively. There is little information on the detection and distribution of bovine Theileria species in northern China. Therefore, this study provides important data for understanding the epidemiology of Theileria species and designing appropriate approaches for the diagnosis and control of bovine theileriosis in northern China.