Calcium channel blocker diltizem transiently inhibits migration and up-regulates metadherin expression in hepatocellular carcinoma cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of calcium channel blocker diltizem in reversing multi-drug resistance (MDR) and on metadherin expression in hepatocellular carcinoma cells and explore the molecular mechanism.@*METHODS@#Hepatocellular carcinoma MHCC97H and 7402 cells were treated with diltiazem hydrochloride, a calcium channel blocker (0, 25, 50, 100, 200, and 400 μmol/L), for 12, 24, or 48 h. Wound healing assay was employed to assess the changes in the mobility and migration of the cells following the treatments, and the changes in the expression levels of metadherin mRNA and protein and P-gp protein were determined using RT-PCR and immunocytochemistry.@*RESULTS@#Diltiazem hydrochloride could transiently inhibit the migration and movement of MHCC97H and 7402 cells in a time-and concentration-dependent manner ( < 0.05). Diltiazem hydrochloride at different concentrations also transiently up-regulated the expressions of metadherin mRNA and protein but did not inhibit the expression of P-gp protein in MHCC97H and 7402 cells.@*CONCLUSIONS@#Calcium channel blocker can transiently inhibit the migration of hepatocellular carcinoma cells and up-regulate the expression of metadherin mRNA and protein through a feedback mechanism, suggesting the potential risk of calcium channel blockers for promoting tumor progression during the treatment of malignant tumors.

Generation of a novel HBeAg transgenic mice using CRISPR/Cas9 technique / 南方医科大学学报

OBJECTIVE@#To generate a new strain of HBeAg transgenic mice using CRISPR/Cas9 technique.@*METHODS@#Hepatitis B virus (HBV) HBeAg gene was cloned and inserted in the pliver-HBeAg expression frame at the site of Rosa26 gene using CRISPR/Cas9 and homologous recombination techniques to construct the pliver-HBeAg expression vector containing HBeAg gene. The linear DNA fragment containing HBeAg gene was obtained by enzyme digestion. Cas9 mRNA, gRNA and the donor vector were microinjected into fertilized eggs of C57BL/6J mice, which were then transplanted into the uterus of C57BL/6J female surrogate mice to obtain F0 generation mice. The F0 generation mice were identified by long fragment PCR to obtain F0 transgenic mice with HBeAg gene. The positive F0 generation mice were bred with wild-type C57BL/6J mice to produce the F1 mice, which were identified by PCR and sequencing. The positive F1 transgenic mice carrying HBeAg gene were backcrossed until the homozygous offspring transgenic mice were obtained. The genotypes of the offspring mice were identified. The expressions of HBeAg and HBeAb in the heterozygous and homozygous HBeAg transgenic mice were detected by automatic chemiluminescence immunoassay, immune colloidal gold technique and immunohistochemistry method.@*RESULTS@#A total of 56 F0 mice were obtained, and 2 of them carried homologous recombined HBeAg gene. Six positive F1 mice were obtained, from which 22 homozygous and 29 heterozygous F2 generation HBeAg transgenic mice were obtained. High concentration of HBeAg protein was detected in the peripheral blood of all the positive HBeAg transgenic mice without HBeAb expression. HBeAg expression was detected in the hepatocytes of HBeAg transgenic mice.@*CONCLUSIONS@#We obtained a new strain of HBeAg transgenic mice with stable expression of HBeAg in the hepatocytes and immune tolerance to HBeAg using CRISPR/Cas9 technique, which provide a new animal model for studying HBV.

Reasons for clinical failure after endovascular treatment of chronic iliac vein occlusions / 中华普通外科杂志

Objective To discuss the risk factors of iliaofemoral vein restenosis after percutaneous recanalization of post-thrombotic occlusion.Method From March 2006 to December 2014,44 limbs in 41 patients with chronic post-thrombotic iliaofemoral vein occlusion underwent endovascular treatment in our hospital.Post-operative patency was evaluated by ultrasonography and anterograde veinography.Results Initial technical success was 90.2％ (37/41).During the follow-up,5 cases of reobstruction were determined,2 were caused by insufficient stent extension into the inferior vena cave.Failure in two cases of intra-stent thrombosis were found due to arbitrary discontinuity of oral anticoagulative therapy.Moreover,2 of 9 cases with venous ulcers in lower extremities were exacerbated after failure of endovascular treatment,we therefore performed surgical ligation and endovenous laser treatment of circumambient perforating veins.Conclusion Insufficient lesion cover,not enough extension to interior vena cave and unreasonable post-operative anticoagulation discontinuity are major causes of intra-stent reobstruction in iliaofemoral venous occlusion.

Clinical effect analysis in the treatment of acute limb arterial critical ischemia / 国际外科学杂志

Objective To discuss the therapeutic effect in the treatment of the acute limb arterial critical ischemia.Methods Collect thirty-nine cases of acute limb arterial critical ischemia in Renji Hospital from Janary 2014 to July 2016.According to the patients' manifestation,these operations were porfermed including thrombectomy,cathetery-directed thrombolysis,mechanical suction bolt,percutaneous angioplasty and stenting.The effect and complications were observed.Results The eighteen patients in 39 cases (46.2％) were dead,including 5 cases without operation,13 operation.The eight cases were amputated during 34 cases of operations.In the 21 out-patients safely,2 cases were not followed up.The time of follow-up was from 3 to 27 months,on average 14.3 months.During the 21 patients,5 cases died from heart cerebrovascular or tumor diseases,3 cases with footdrop,2 cases with toe amputations,3 cases with distal leg and foot anesthesias.Conclusions The patiens with acute limb arteries critical ischemia must be treated as early as,and reinforced the management of multiple organ function,which may improve the diseases' therapeutic effect.

Percutaneous endovascular treatment of below-the-knee arterial bifurcation lesions with coronary bifurcation stenting techniques / 中华普通外科杂志

Objective To report the preliminary results of drug eluting stents (DES) in infrapopliteal bifurcation lesions after failed angioplasty.Methods From April 2011 to December 2013, 21 patients with critical limb ischemia and infrapopliteal atherosclerotic disease underwent DES placement in tibial bifurcation using 4 techniques: balloon and stent (single stent), T-shape double-stent, Crush double-stent and Culotte double-stent.Clinical and angiographic end points included: Amputation-free survival, target lesion revascularization (TLR)-free survival, target vessel revascularization (TVR)-free survival.Angiographic primary and secondary patency of DES, 2-vessel primary patency (2VPP), and 1-vessel primary patency (1VPP).Results Technical success was achieved in all patients (100％) without major intraoperative complications.ABI increased from 0.23 ± 0.07 to 0.67 ± 0.24 (P ＜ 0.05) after 1 week.The mean follow-up intervals were 23 ± 7 months;amputation-free survival, TLR-free survival and TVR-free survival were 100％, 90.5％ and 79.1％, respectively, at 1 year.8 patients underwent angiography reevaluation.The angiographic primary and secondary patency of DES were 62.5％, 87.5％, respectively.The primary patency of 2VVP and 1VVP were 28.6％, 57.1％, respectively at 1 year.Conclusions Bifurcation stenting techniques utilized in coronary arteries can be used in the infrapopliteal arteries.Although provisional stenting in the origin of tibial bifurcation cannot inhibit restenosis in distal lesions treated with balloon angioplasty, clinical outcomes and limb salvage rates are acceptable for these techniques.

A novel histological typing and grading-scale system of colorectal cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To formulate a novel histological typing and grading-rated system for colorectal cancer (CRC) for evaluating the biological behavior of CRC and prognosis.</p><p><b>METHODS</b>According to the highly heterogeneous histological features, WHO classification and histological differentiation criteria, and other biological behavior parameters of CRC, a novel histological typing and grading-scale system for CRC was designed. The histological typing and corresponding grading-scale of CRC was defined as the following: (1) No mucin-producing adenocarcinoma, including tubular adenocarcinoma, sieve-like acne adenocarcinoma, medullary carcinoma, serrated adenocarcinoma and micropapillary carcinoma, etc. (1-3 points); (2) Mucin-producing adenocarcinoma, including mucinous adenocarcinoma and signet ring cell carcinoma (3-4 points); (3) Squamous cell carcinoma (1-3 points); (4) Neuroendocrine tumors, including neuroendocrine tumors, neuroendocrine carcinoma (1-4 points); (5) The special type of CRC, including clear cell carcinoma, spindle cell carcinoma, etc. (4-points); (6) Undifferentiated carcinoma (5 points). The pathology report form was formatted based on the major histological type with the secondary histological type. The final total score of CRC was defined as the sum of the corresponding grading scores for different histological types. The total score of a single-structure CRC was defined as the corresponding grading score multiplied by 2. A total of 666 patients with advanced CRC were pathologically reviewed and analyzed to assess the correlation of the histological typing and grading scores with TNM staging and lymph node metastasis.</p><p><b>RESULTS</b>The results showed a significant correlation of the histological grading-scale and TNM staging and lymph node metastasis (P<0.05). The scores of CRC histological grading-scale increased synchronously with the TNM staging and lymph node metastasis rate.</p><p><b>CONCLUSION</b>The novel histological grading system allows objective evaluation of the biological behaviors and prognosis of CRC for determining individualized postoperative treatment. This system still needs further revision and updates based on evidence from prospective, multi-centered, large-scale trials.</p>

Antibacterial mechanism and safety of zinc oxide / 中国组织工程研究

BACKGROUND:Zinc oxide has shown extraordinary antibacterial effect. But the antibacterial mechanisms of zinc oxide have not been wel developed, and the safety of zinc oxide has not yet been confirmed. OBJECTIVE:To summarize the antibacterial mechanism and safety of zinc oxide based on the current research situation of zinc oxide. METHODS:The CNKI database and PubMed database (2005-01/2013-08) were used to search the related articles about zinc oxide antibacterial mechanism and safety. The retrieval keywords were“zinc oxide or ZnO, antibacterial”,“zinc oxide or ZnO, safety”both in English and Chinese. RESULTS AND CONCLUSTION:The antibacterial mechanism of zinc oxide is complex. Zinc oxide reveals its antibacterial power by photo catalysis, zinc ion dissolve out, and active oxygen. But the relationship between photo catalysis and active oxygen is different based on different studies. The safety of zinc oxide is not confirmed, and different studies have different results. Especial y for the nano-zinc oxide, some studies have showed that the zinc oxide is toxic. Al of these mean much more researches are needed to verify the safety of zinc oxide.

Endovascular treatment of chronic total occlusions of the iliac vein / 中华普通外科杂志

Objective To present clinical results of endovascular treatment of total iliac vein occlusions and to discuss the technique details of this treatment. Methods From Feb 2006 to Aug 2010,15 patients with chronic total occlusive lesions of the iliac vein and adjacent vein segments underwent endovascular treatment. Average age was (62 ±7) years (range 35 to 81 years), the male/female ratio was 12: 3. Clinical score of CEAP was grade 3 in 33. 3%, grade 4 in 40%, grade 5 in 13. 3%, and grade 6 in 13.3%. Venography showed left iliac vein was occluded in all 15 patients, common femoral vein was occluded in 14 patients, and superior femoral vein was occluded in 9 patients, however profounder femoral vein was patent in these patients with superior femoral vein being occluded. Results No postoperative major morbidity or mortality was seen. The technique success rate was 93. 3%. Treatment failure was caused in one case for a wrong selection of the femoral vein approach site. The average length of stent was 18. 4 cm.In 12(80. 0% ) stents crossed the inguinal ligament. The average follow-up time was 11.6 ± 2. 4 months.The primary patency rates of the stents at 6 months were 92. 9%. 10 (66. 7% ) patients were asympotomatic, 3(20% )were improved, 1 (6. 7% ) was unchanged, and 1 (6.7%) was worse, compared to before intervention. Conclusions Endovscular recanalization and stent placement is a safe and effective treatment for chronic total occluded iliac veins, with good patency, significant symptom resolution, and minimal morbidity in the short term follow-up.

Transcatheter thrombolysis via the small saphenous vein for acute lower limb deep venous thrombosis / 中华普通外科杂志

Objective To evaluate transcatheter thrombolysis via the small saphenous vein in the treatment for lower limb acute mix-deep venous thrombosis. Method From Jan 2005 to Mar 2007.37 patients with lower limb acute mix-DVT underwent catheter-directed thrombolysis via the small saphenous vein with urokinase(149 ±71)×104 IU continuous infusion.The venous patency score and the rate of patency improvement were observed by venograms before and after therapy.Twenty-two patients were followed up for(12 ±4)months. Results Venous patency score were significantly improved(Z=-5.330,P＜0.01).The mean rate of venous patency was 50.17%±15%,and there was no complication.Venogram on 6～12 month follow up showed a venous patency of 58%±13%(Z=-3.545,P＜0.01),and that on 13～18 months was 68%±20%(Z=-2.201,P＜0.05). Conclusion This preliminary experience suggests that catheter-directed thrombolysis via the small saphenous vein with urokinase for acute lower limb mix-DVT iS safe and effective.

The measurement and clinic significance of tissue Inhibitors of metalloproteinase-1 on children with myocardltis / 中国基层医药

Objective To evaluate the relationship between serum tissue inhibitors of metaUoproteinase-1 and children with myocarditis,and provide evidences to support the measurement of serum tissue inhibitors of metallopro- teinase-1. Methods Tissue inhibitors of metalloproteinaee-1 and bioehem indexes of the harvested specimens and e -chocardiogram were measured from children with myocarditis(age: 1～14 years old, n = 9) and healthy young chil- dren(age: 1～14 years old, n = 9). Results Children with myocarditis showed reduced ejection fraction and dilated left ventricular compared with healthy children. Tissue inhibitors of metalloproteinaee-1 in children with myocarditis was significantly reduced compared with healthy children. Aspartie acid transanainase, lactate dehydrogenase,creatine kinase, hydroxybutyrie acid dehydrogenase, MB isoenzyrne of creatine kinase, cardiac troponin Ⅰ , C reactive protein showed no difference between two groups. Conclusion The reduced serum level of tissue inhibitors of metallopro- teinase-1 may show the severity of children with myocarditis and maybe one of the mechanisms of children with di- lated cardiomyopathy following myocarditis.

Intraluminal venous catheter-directed thrombolysis for lower limb acute deep venous thrombosis / 介入放射学杂志

Objective To evaluate the efficacy of intraluminal catheter-directed thrombolysis in treatment of lower limb acute deep venous thrombosis(DVT).Methods Thirty-six consecutive patients with lower limb acute DVT underwent intraluminal cathter-directed thrombolysis with urokinase continuous infusion immediately.The circumferences of the normal and affected limbs were measured before and after lysis,the venous patency scores and the rates of patency improvement were observed by venograms,together with follow up record after 6 months.Results After lysis,the circumferencial differences in thigh and calf showed significant difference(P

Anti-tumor immunity induced by mouse lung cancer B7 vaccine / 中国肺癌杂志

<p><b>BACKGROUND</b>To study the cell immunity induced by lung cancer B7 vaccine, FLB2C cells.</p><p><b>METHODS</b>Compared withparental lung cancer cell LA795 line, proliferation of mouse T lymphocytes stimulated by FLB2C was observed through mixed lymphocyte culture. CTLL cell MTT test was used to detect whether FLB2C stimulated T lymphocyte to secrete IL-2. After immunized with the FLB2C and LA795 cells, the CTL activity of mouse was observed in vivo.</p><p><b>RESULTS</b>The spleen lymphocyte proliferation stimulated by FLB2C cells was remarkably stronger than that by LA795 . FLB2C might stimulated the T lymphocyte to secrete IL-2 in vitro, but LA795 didn't. FLB2C cell could induce CTL activity in vivo and the induced CTL killed FLB2C cells and LA795 in vitro, with the killing rate of 34% and 25.3% respectively; while LA795 induced CTL killing rate being 10.5% and 12.25% respectively (P < 0.01).</p><p><b>CONCLUSIONS</b>FLB2C can stimulate T cell immunity in vivo or in vitro. The effect of FLB2C is significantly stronger than that of LA795 . The results suggest that FLB2C may be used to treat lung cancer through improving immunity. This provides immunological basis for applying FLB2C as a vaccine to clinical use.</p>

Establishment and characteristics of multidrug-resistant Lewis lung cancer cell line / 中国肺癌杂志

<p><b>BACKGROUND</b>To establish an ADM resistant Lewis lung cancer cell line and to investigate its biological characteristics.</p><p><b>METHODS</b>The multidrug-resistant cell line was gradually induced by ADM from Lewis lung cancer cell line (L3-8), its growth characteristics and cancinogenicity were observed. The fluorescent density of ADM and Rh-B in the cells were analyzed by IPP image analysis system. The ADM concentration in cells was assayed with fluorescent meter. The IC50 was evaluated by MTT assay and the drug resistant index was counted.</p><p><b>RESULTS</b>The drug resistant Lewis lung cancer cell line, L3-8/ADM, which can grow in the medium containing 0.4mg/l ADM was acquired after 14 months selective culture. Its tumor generatic rate was 10/10. When the L3-8/ADM was cultured in 0.15mg/l ADM and general 1640 medium, the doubling time was 22.0h and 21.8h separately. After culturing in 4mg/l ADM and Rh-B medium, the fluorescent density ratio of L3-8/ADM and L3-8 were 2.82:1 and 2.65:1 separately. The ADM concentration of L3-8/ADM was only 64.7% of its parental, but there was no significant difference in their ADM release rate. Except for ADM, L3-8/ADM was resistant to DNR, VCR, MIT and CDDP in different degrees.</p><p><b>CONCLUSIONS</b>L3-8/ADM cell is a typical MDR cell line. The cell membrane obstruction of drug infiltration might be the cause of drug resistance. This study will provide a basis for the establishment of lung cancer MDR animal model.</p>

Treatment of lower extremty deep venous thrombosis in 165 patients / 中国普通外科杂志

Objective To evaluate the therapeutic results of embolectomy and thrombolysis for lower extremity deep vein thrombosis in 165 patients. Methods Embolectomy was undertaken with Fogarty catheter in 64 patients while 101 patients were treated by thrombolytic agents. Results Ninety-six cases (58.2%) were followed-up, ranging from 7 months to 218 months with an averge of 71 months including 44 cases undergoing embolectomy and 52 cases receiving thrombolytic agents. Symptoms disappeared in 20.5% of patients in embolectomy group and in 13.5% in thrombolytic group.In embolectomy group,the superficial varicoses and derma pigmentation in lower extremity appeared in 31.8% of patients,while in thrombolytic group the signs appeared in 50.5% of patients, respectively.Conclusions Embolectomy protecting the valve of deep trunk vein in some patients helps to reduce later complications of lower extremity deep venous thrombosis.

Catheter-directed thrombolysis for lower limb deep venous thrombosis / 介入放射学杂志

Objective To evaluate the efficacy of catheter-directed thrombolysis in the treatment of lower limb acute deep venous thrombosis(DVT). Methods One hundred and four patients with lower limb acute DVT underwent cathter-directed thrombolysis with continuous infusion of urokinase(154.27 ? 76.31 ? 104 IU). Fourteen pationts were implanted with stents for the residual stenoses. The circumferences between normal and affected limbs were measured before and after the thrombolysis. The venous patency score, the rate of patency improvement were eveluated by venography and the patients were followed up for six months. Results After thrombolysis, the venography revealed that venous patency improved in 92 patients(mean 52.42% ? 16.37%, P