Objective:To discuss the effect of D-limonene on the proliferation and apoptosis of the glioblastoma(GBM)cells,and to clarify its possible mechanism.Methods:The GBM cells were divided into control group(0 mmol·L-1 D-limonene)and 0.2,0.4,0.6,0.8,and 1.0 mmol·L-1 D-limonene groups.CCK-8 method was used to detect the inhibitory rates of proliferation of the cells in various groups;clone formation assay was used to detect the clone formation rates of the cells in various groups;Annexin Ⅴ-FITC/PI method was used to detect the apoptotic rates of the cells in various groups;Western blotting method was used to detect the expression levels of protein kinase B(AKT),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and poly adenosine diphosphate(ADP)-ribose polymerase(PARP)proteins in the cells in various groups;imunofluorescence method was used to detect the expression levels of cleaved Caspase-3 protein in the cells in various groups.Fifteen model mice with subcutaneous tumor xenografts were randomly divided into blank group(0 mg·kg-1·d-1 D-limonene),low dose of D-limonene group(200 mg·kg-1·d-1 D-limonene),and high dose of D-limonene group(400 mg·kg-1·d-1 D-limonene),and there were 5 mice in each group.The inhibitory rates of the tumor in vitro in various groups were calculated;HE staining and immunohistochemical staining were used to observe the morphology of subcutaneous tumor tissue of the mice in various groups and the growth curves of the tumor were drawn;immunohistochemical assay was used to detect the positive expression rates of Ki67 protein in subcutaneous tumor tissue of the mice in various groups;TUNEL staining was used to detect the apoptosis of the tumor cells in various groups.Results:In control group,the cells were spindle-shaped,in good condition,growing closely and adherently,with normal organelles and cytoplasm.After treated for 48 h,the cells in 0.6 mmol·L-1 D-limonene group showed reduced volume,intact but more permeable cell membranes,shrunken cytoplasm,internal vacuole structures,and some fragments floating in the solution.The cells in 0.8 and 1.0 mmol·L-1 D-limonene groups exhibited significant apoptotic bodies and were in an apoptotic state.The CCK-8 results showed that compared with control group,the inhibitory rates of proliferation of the U87,LN229,and GL261 cells in 0.6,0.8,and 1.0 mmol·L-1 D-limonene groups were significantly increased(P<0.01),the inhibitory rates of proliferation of the U87 and GL261 cells were significantly increased(P<0.01).The clone formation assay results showed that compared with control group,the clone formation rates of the U87,LN229,and GL261 cells in 0.4,0.6,and 0.8 mmol·L-1 D-limonene groups were significantly decreased(P<0.05 or P<0.01).The AnnexinⅤ-FITC/PI results showed that compared with control group,after treated with D-limonene for 48 h,the apoptotic rates of the LN229 cells in 0.6,0.8,and 1.0 mmol·L-1 D-limonene groups were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of Bax proteins in the LN229 cells in 0.6,0.8,and 1.0 mmol·L-1 D-limonene groups were significantly increased(P<0.01),while the expression levels of AKT and Bcl-2 proteins were significantly decreased(P<0.01),the expression level of PARP protein in the LN229 cells in 0.8 and 1.0 mmol·L-1 D-limonene group was significanthy increased(P<0.01).The immunofluorescence results showed that compared with control group,the expression levels of cleaved Caspase-3 protein in the LN229 cells in 0.6,0.8,and 1.0 mmol·L-1 D-limonene groups were significantly increased(P<0.01).Compared with blank group,the tumor volumes of the mice in low and high doses of D-limonene groups were significantly decreased(P<0.01).Compared with blank group,the tumor weights of the mice in low and high doses of D-limonene groups were significantly decreased(P<0.05),and the inhitory rates of tumor were significantly increased(P<0.05).The tumor cells in blank group were diffusely distributed,with deepened nuclear staining and increased nucleocytoplasmic ratio;a large number of degenerated and necrotic tumor cells were observed in tumor tissue of the mice in low and high doses of D-limonene groups.Compared with blank group,the positive expression rates of Ki67 protein in tumor tissue of the mice in low and high doses of D-limonene groups were significantly decreased(P<0.01).Compared with blank group,the apoptotic rates of tumor cells of the mice in low and high doses of D-limonene groups were significantly increased(P<0.01).Conclusion:D-limonene has the inhibitory effect on the proliferation of the GBM cells;its mechanism may be related to the regulation of AKT protein expression and the activation of the Caspase-3 pathway to induce the apoptosis.

Objective:To establish the common carotid artery aneurysm models of Wallstent double stent overlapping implantation in miniature pigs, and evaluate the safety and effectiveness of this procedure by observing the imaging and pathological changes.Methods:Sidewall aneurysm and fusiform aneurysm models in Bama miniature pigs were established surgically and 2 Wallstent stents were overlapped and implanted in situ. Aneurysm healing immediately after surgery and during 8 weeks of follow-up were evaluated according to 2D-DSA by O'Kelly-Marotta (OKM) grading scale and Kamran scale; degrees of stent adhesion immediately after surgery and status of stent endothelialization and aneurysm healing at 2, 4, and 8 weeks after surgery were observed by high resolution C-arm CT(HR-CBCT) and optical coherence tomography (OCT); and the changes of stent endothelialization were evaluated by comparing the HR-CBCT and OCT results with histopathology at 8 weeks after surgery. Perioperative adverse events were recorded.Results:After successful establishment of common carotid artery aneurysm models (including 4 sidewall aneurysms and 4 fusiform aneurysms with average diameter of [11.0±2.8] mm) in 8 miniature pigs, a total of 16 Wallstent stents (2 in each aneurysm) were implanted across the aneurysmal neck, with a technical success rate of 100%. No serious complications such as acute stent thrombosis, or aneurysm rupture and bleeding were observed in the perioperative period. The 2D-DSA immediately after surgery showed obvious intracranial contrast agent retention in 6 patients (1 patient in grading 1, 3 in grading 2, and 2 in grading 3) and aneurysm occlusion in 2 patients (grading 4). Eight weeks after follow-up, all 8 aneurysms had complete occlusions (grading 4); and 2 experimental pigs had in-stent restenosis, with stenosis rates of 52% and 67%, respectively. HR-CBCT and OCT immediately after surgery and during follow-up indicated that the stent metal braid was gradually covered by proliferating intima, with disappeared aneurysm. The cause of in-stent restenosis in 2 experimental pigs was local intima hyperplasia resulted from poor stent adhesion, and pathological findings indicated that the intima hyperplasia was mainly composed of smooth muscle cells and fibrous connective tissues.Conclusion:In animal models, Wallstent stent overlapping implantation is safe and effective in common carotid aneurysms, but intraoperative adverse adhesion of overlapping stent should be avoided.

Objective    To summarize the clinical experience of surgical resection of synchronous multiple ground-glass nodules (SMGN), and explore the individualized diagnosis and treatment strategy of SMGN. Methods    Clinical data of 84 patients with SMGN who underwent thoracic surgery in Anhui Chest Hospital from July 2016 to August 2018 were analyzed retrospectively, including 18 males (21.4%) and 66 females (78.6%), aged 32-80 (55.6±10.3) years. The results of operation and the information of GGNs were analyzed. Results    Except for 1 patient who was converted to thoracotomy due to extensive dense adhesion of thoracic, other patients underwent video-assisted thoracoscopic surgery successfully. All patients recuperated successfully after operation, without severe perioperative complications or death. Finally, 79 patients were diagnosed as malignant tumors (94.0%), and 5 patients of benign lesions (6.0%). A total of 240 GGNs were removed, among which there were 168 pGGNs, including 68 benign lesions (40.5%) and 100 malignant tumors (59.5%), and 72 mGGNs, including 2 benign nodules (2.8%) and 70 malignant tumors (97.2%). Nodules diameter (P<0.001), consolidation/maximum diameter of nodule ratio (P<0.001), vacuole sign (P<0.001), air bronchograms sign (P=0.001), spine-like process (P=0.001), pleural indentation sign (P<0.001), lobulation sign (P<0.001), and vascular convergence (P=0.002) were correlated with malignant tumor. Conclusion    Analysis of the imaging features of GGNs by thin-section CT scan and three-dimensional reconstruction is of great value in predicting the benign and malignant nodules, which can guide the surgical decision-making and preoperative planning. Through reasonable preoperative planning and following certain principles, simultaneous surgical treatment for SMGN is safe and feasible.

Objective:To investigate pouch function, quality of life, and their influencing factors after ileal pouch-anal anastomosis (IPAA) in patients with ulcerative colitis (UC) .Methods:Clinical data of 111 patients undergoing IPAA in the UC database at Department of General Surgery, Jinling Hospital from 2014 to 2020 were retrospectively analyzed. Through the questionnaire, pouch functional score (PFS) and Cleveland global quality of life (CGQL) were obtained. These patients were diveided into two groups according to the PFS score. Univariate and multivariate analysis were used to compare the clinical features and reveal the risk factors of PFS.Results:A total of 111 patients were enrolled in this study, including 65 males and 38 females. 63 patients were in the good pouch function group, and 48 in the poor pouch function group. The median score of PFS was 7 (3.0-10.0) . Univariate analysis showed that compared with the good pouch function group, the poor pouch function group had a longer duration of disease before operation[44.5 (21.0-86.0) months vs. 14.0 (5.0-70.0) months, P = 0.005], a shorter follow-up time[16.0 (7.0-28.8) months vs. 26.0 (15.0-39.0) months, P = 0.020], and a higher proportion of pouchitis (29.1% vs. 12.9%, P = 0. 031) . The multivariate logistic regression analysis showed duration of disease before operation ( OR = 0.974, 95% CI: 0.951-0.997, P = 0.030) and pouchitis ( OR = 3.251, 95% CI: 1.168-9.045, P = 0.024) were the independent risk factor of PFS in UC patients.The median CGQL was 0.77 (0.67-0.87) . The median surgical satisfaction score was 9.0 (8.0-10.0) . There was a correlation between CGQL and PFS ( rs = -0.376, P<0.001) . Conclusions:Most patients recover well after IPAA, and are highly satisfied with the operation. A good pouch function has a positive influence on the quality of life.

Objective:To investigate pouch function, quality of life, and their influencing factors after ileal pouch-anal anastomosis (IPAA) in patients with ulcerative colitis (UC) .Methods:Clinical data of 111 patients undergoing IPAA in the UC database at Department of General Surgery, Jinling Hospital from 2014 to 2020 were retrospectively analyzed. Through the questionnaire, pouch functional score (PFS) and Cleveland global quality of life (CGQL) were obtained. These patients were diveided into two groups according to the PFS score. Univariate and multivariate analysis were used to compare the clinical features and reveal the risk factors of PFS.Results:A total of 111 patients were enrolled in this study, including 65 males and 38 females. 63 patients were in the good pouch function group, and 48 in the poor pouch function group. The median score of PFS was 7 (3.0-10.0) . Univariate analysis showed that compared with the good pouch function group, the poor pouch function group had a longer duration of disease before operation[44.5 (21.0-86.0) months vs. 14.0 (5.0-70.0) months, P = 0.005], a shorter follow-up time[16.0 (7.0-28.8) months vs. 26.0 (15.0-39.0) months, P = 0.020], and a higher proportion of pouchitis (29.1% vs. 12.9%, P = 0. 031) . The multivariate logistic regression analysis showed duration of disease before operation ( OR = 0.974, 95% CI: 0.951-0.997, P = 0.030) and pouchitis ( OR = 3.251, 95% CI: 1.168-9.045, P = 0.024) were the independent risk factor of PFS in UC patients.The median CGQL was 0.77 (0.67-0.87) . The median surgical satisfaction score was 9.0 (8.0-10.0) . There was a correlation between CGQL and PFS ( rs = -0.376, P<0.001) . Conclusions:Most patients recover well after IPAA, and are highly satisfied with the operation. A good pouch function has a positive influence on the quality of life.

Through the effort in nearly 40 years of Tanzania, the total number of reported cases, the prevalence of HIV/AIDS and the number of death have declined, while the number of patients receiving antiretroviral therapy has increased significantly in Tanzania. At the same time, however, there are new challenges that require more attention, such as the HIV spread to rural areas, middle and lower social classes, and female teenagers. Although the overall performance of the HIV/AIDS prevention and control response is great, there is still a need to survey the new trends of this disease at the micro-level in Tanzania. In addition, there is a necessity of interdisciplinary concern due to the prevalence of heterosexual anal sex in Tanzania.

Perfluorinated compounds (PFCs), a group of persistent organic pollutants, have been widely detected in environmental media and posed great threat to human health. The researches on environmental pollution and health concern of PFCs are the hotspot areas. Because PFCs contain lots of homologs and isomers which are detected at trace levels (ng/g or μg/L) in environment, advanced and reliable analytical methods for determination of PFCs in environment are urgently needed. At present, studies on analytical methods of trace PFCs in environmental samples have been widely carried out in China and abroad. However, systematic review on the sample pretreatment, analytical method, and matrix effect of PFCs determination in complex environmental matrixes is relatively scarce. Therefore, this paper reviews the pretreatment methods, martix effects, and detection techniques (especailly isomers) of PFCs in environment samples (water, sediment/sluge, soil and plant). We hope that this review may provide valuable reference for the enviromental researches on PFCs.

Objective To investigate the effect of HBP-A on meniscal injuries and the expressions of genes associated with pathological hypertrophy and calcification of the meniscusinduced by abnormal loading. Methods Bovine meniscus explants were subjected to 25%strain at 0.3 Hz for 3 h and treated with 0.6 mg/mL of HBP-A. The cell viability in the meniscus explants after 72 hin culture was determined using live/dead staining and the expression levels of genes associated with pathological hypertrophy and calcification of the meniscus (ANKH, ENPP1, ALP, MMP13, and IL-1) were measured using real-time PCR and Western blotting. The conditioned medium was collected for testing sulfated glycosaminoglycan (GAG) release. Results The number of dead cells, loss of proteoglycan content, and the expressions of ANKH, ENPP1, ALP and MMP13, and IL-1 at both the mRNA and protein levels were all significantly lower in the meniscus explants treated with 0.6 mg/mL HBP-A than in the explants with only 25%abnormal pressure stimulation (n=3, P<0.05). Conclusion HBP-A can effectively alleviate meniscal injuries induced by abnormal loading and suppress the expressions of genes related with pathological hypertrophy and calcification of the meniscus, and can serve as a potential drug for treatment of knee osteoarthritis.

Objective To investigate the effect of HBP-A on meniscal injuries and the expressions of genes associated with pathological hypertrophy and calcification of the meniscusinduced by abnormal loading. Methods Bovine meniscus explants were subjected to 25%strain at 0.3 Hz for 3 h and treated with 0.6 mg/mL of HBP-A. The cell viability in the meniscus explants after 72 hin culture was determined using live/dead staining and the expression levels of genes associated with pathological hypertrophy and calcification of the meniscus (ANKH, ENPP1, ALP, MMP13, and IL-1) were measured using real-time PCR and Western blotting. The conditioned medium was collected for testing sulfated glycosaminoglycan (GAG) release. Results The number of dead cells, loss of proteoglycan content, and the expressions of ANKH, ENPP1, ALP and MMP13, and IL-1 at both the mRNA and protein levels were all significantly lower in the meniscus explants treated with 0.6 mg/mL HBP-A than in the explants with only 25%abnormal pressure stimulation (n=3, P<0.05). Conclusion HBP-A can effectively alleviate meniscal injuries induced by abnormal loading and suppress the expressions of genes related with pathological hypertrophy and calcification of the meniscus, and can serve as a potential drug for treatment of knee osteoarthritis.

Objective To explore the efficacy and safety of percutaneous transhepatic portal vein or transjugular intrahepatic portosystemie shunt (TIPS)to implant the portal vein metallic stent in treatment of cavernous transformation of portal vein (CTPV).Methods Clinical and imaging data of 8 patients with CTPV were retrospectively analyzed who were treated in our hospital.All patients were treated with metallic stent implantation in portal vein including 3 patients by TIPS and 5 by percutaneous transhepatic portal vein.Results All patients were successful in the stent implantation without any occurrence of serious complications such as intra-abdominal hemorrhage and so on.Intraoperative angiography showed blood circulated freely in these stents.1 day-2 weeks later,the patients symptoms of abdominal pain and gastrointestinal bleeding were obviously relieved or disappeared.Follow up 1 month-3 years,1 patient with stent occlusion after one year of operation,the blood flow recovery after stent reimplantation,and the remaining patients,color doppler ultrasound reflected patency of blood flows in their stents.No one suffered from gastrointestinal bleeding or abdominal pain again.Conclusion Implantation of portal vein metallic stent via percutaneous transhepatic portal vein or via TIPS in treatment of cavernous transformation of portal vein is safe and effective.