Polyamidoamine-polyethylene glycol (PAMAM-PEG) copolymers were synthesized using IPDI as coupling reagent by two-step method. The copolymers were characterized by IR spectrum and 1H NMR spectrum, and the PEG conjugating ratios of the copolymers were calculated equal to 10% and 30% separately. MTT assay indicated that after PEGylation a lower cytotoxicity of the copolymers could be found, and with increasing PEG conjugating ratio the cytotoxicity decreased obviously. Agarose gel retardation assay demonstrated that PAMAM-PEG copolymers could be combined with DNA and PAMAM-PEG/DNA complexes were prepared by self-assembly. DLS measurement showed that when N/P > or = 50, the particle size of copolymer/ gene complexes was in a range of 150-200 nm, and the zeta potential was in a range of 10-25 mV. In vitro gene transfection illustrated that when N/P < or = 50, the gene transfection efficiency of PAMAM-PEG copolymers was a little less than that of PAMAM-G5, but the transfection efficiency can be raised by increasing N/P ratio or transfection time. Considering both cytotoxicity and transfection efficiency aspects PAMAM-PEG-13 was more effect than PAMAM-PEG-39 in PEGylation.
Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Survival ; drug effects ; DNA ; chemistry ; pharmacology ; Dendrimers ; chemical synthesis ; pharmacology ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Isocyanates ; chemistry ; Liver Neoplasms ; pathology ; Particle Size ; Polyamines ; chemistry ; Polyethylene Glycols ; chemical synthesis ; chemistry ; pharmacology ; Transfection

OBJECTIVETo investigate the therapeutical effect and side-effect of docetaxel combined with cisplatin (DDP) on the treatment of local advanced esophageal cancer with concomitant radiation therapy.

METHODSNinety patients with LOCAL advanced esophageal squamous cell carcinoma were divided into two groups: (DDP + 5-Fu) group and (docetaxel + DDP) group. Chemotherapy was carried out every 4 weeks for a total of 4 courses. The radiation dose was 50.4 Gy/28FX.

RESULTSThe median survival time of patients in the (DDP + 5-Fu) group was 16 months and that in (docetaxel + DDP) group was 21 months (P = 0.0278). The 3-year survival rate in the (docetaxel + DDP) group was obviously higher than that in the (DDP + 5-Fu) group (23.9% vs. 12.1%). The ORR in (docetaxel + DDP) group (84.5%) was significantly higher than that in the (DDP + 5-Fu) group (71.1%) (P = 0.025). No significant differences were observed in the incidence of side-effects in the two groups.

CONCLUSIONSThe conventional dose chemotherapy of docetaxel + DDP with concomitant radiation therapy showed a better partial remission rate and long-term survival rate for the treatment of local advanced esophageal cancer than the traditional chemotherapy (DDP + 5-Fu) with concomitant radiation therapy and the side-effects are not increased.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; drug therapy ; pathology ; radiotherapy ; Cisplatin ; administration & dosage ; adverse effects ; Combined Modality Therapy ; Cystic Fibrosis ; etiology ; Dose Fractionation ; Esophageal Neoplasms ; drug therapy ; pathology ; radiotherapy ; Female ; Fluorouracil ; administration & dosage ; adverse effects ; Follow-Up Studies ; Humans ; Leukopenia ; chemically induced ; Male ; Middle Aged ; Neoplasm Staging ; Remission Induction ; Survival Rate ; Taxoids ; administration & dosage ; adverse effects ; Young Adult

Objective To evaluate the safety and efficacy of transplantation of purified CD34 + cells (PCCs) in treatment of critical limb ischemia (CLI) caused by thromboangiitis obliterans (TAO).Methods From May 2009 to June 2015,34 TAO-induced-CLI cases underwent PCCs transplantation.None of these patients were eligible for surgical or endovascular revascularization.G-CSF was subcutaneously injected for 5 days before peripheral CD34 + cells were isolated,purified and intramuscularly injected in the limbs.Patients were regularly follow-up.Results Technical success was achieved in all cases.The mean number of transplanted cells was (7.5 ± 2.4) × 105/kg.The follow-up was accomplished in 32 cases,ranging from 6 to 79 months (mean 45 ±24 months),and two patients were lost.Wong-Baker FACES pain rating scale score significantly decreased from 8.0 ±2.0(4-10)to 2.2 ±3.1 (P ＜0.05) at 1 month.The Peak pain-free walking time improved from (4.0 ± 2.0) min to (13.5 ± 5.3) min (P ＜ 0.05) at 3 months and (19.0 ± 3.1) min (P ＜ 0.05) at 6 months.The ankle-brachial index increased from 0.42 ± 0.20 to 0.50 ± 0.10 (P ＜ 0.05) at 3 months and 0.52 ± 0.11 (P ＜ 0.05) at 6 months,respectively.Transcutaneous partial oxygen pressure rose from (25 ± 11) mmHg to (48 ± 11) mmHg(P ＜ 0.05) at 3 months and (58 ± 10) mmHg (P ＜ 0.001) at 6 months,respectively.Ulcers healed in 21 out of 22 patients at (5 ± 4) months.The overall amputation-free survival rate was 94.1％ at 6 months and 91.2％ at 48 months.No major adverse events were observed perioperatively or postoperatively.Conclusions Transplantation of PCCs could yield safe,satisfactory and durable treatment outcomes in patients with TAO-induced-CLI.

Objective To explore the application of rotation map of insulin injection site in patients with type 2 diabetes in-sulin intensive therapy.Methods 240 patients with type 2 diabetes hospitalized in the endocrinology department for insulin intensive therapy were enrolled in the study by random sampling method from February to September 2016.The patients were divided into ob-servation group and control group by random digit table, 120 cases in each group. The routine insulin injection method was used to guide the patients in the control group. The patients in the observation group were guided by the rotation map of insulin-injection sites during the hospital stay.In the three months and six months after discharge,the two groups were compared in terms of cutaneous reaction. Result The cutaneous reaction of the insulin injection site in the two groups was compared,showing that the incidences of cutaneous reaction in the observation group were significantly lower than the control group 3 months and 6 months after discharge(P<0.05). Conclusion The rotation map of insulin injection sites can prevent patients from repeatedly injecting insulin and thus reduce the incidence of cutaneous reaction.It is worthy of clinical application.

Objective To understand the characteristics of sexual network of 321 HIV positive MSM and HIV infection status among sexual partners in Zhejiang Province. Methods A cross-sectional study selected HIV positive MSM diagnosed between September, 2015 and September, 2016 were conducted in Hangzhou, Ningbo, Wenzhou in Zhejiang Province using self-administrated questionnaire. Information related to socio-demographic characteristics, risky behavior, size of sexual network were collected. Results A total of 321 HIV positive MSM were recruited in this study. 71.76％ aged 20 to 40 years old and 51.71％ were native. The rate of self-reported sexual behavior with stable homosexual partners, casual homosexual partners, spouse and other heterosexual partners at investigation were 37.78％, 45.00％, 36.56％ and 66.67％. The rate of never using condom with stable homosexual partners, casual homosexual partners, spouse were 27.91％, 16.36％ and 44.00％. HIV positive rate among stable homosexual partners, casual homosexual partners, spouse and other heterosexual partners were 28.26％ , 3.77％, 11.43％ and 0.00％. Conclusion The size of sexual network was large, especially in casual sexual network. Unsafe sexual behavior, like consistent condom use was low and multiple sexual partners . HIV infection was high among stable homosexual partners and spouse.

OBJECTIVETo determine the incidence of TEL-AML1 fusion gene in childhood acute lymphoblastic leukemia (ALL) and to compare the clinical features between TEL-AML1 positive and negative patients.

METHODSSamples of bone marrow or peripheral blood were collected from 95 newly diagnosed ALL children and the TEL-AML1 fusion gene was detected using nested reverse transcription-polymerase chain reaction (RT-PCR). The ALL patients were stratified into TEL-AML1 positive and negative groups and the clinical features were compared.

RESULTSAmong 95 patients, 20 (21.05%) were TEL-AML1 positive. The median age of TEL-AML1 positive patients was 5.9 years old and M/F ratio was 1.22:1. There were significant differences between TEL-AML1 positive and negative patients in hepatomegaly (2.75 cm vs. 4 cm below costal arch, P=0.006), splenomegaly (0 cm vs. 3 cm below costal arch, P < 0.001), initial white blood cell count (median 7.40 x 10(9)/L vs.18.70 x 10(9)/L, P=0.011), initial peripheral blood blast (median 2.45 x 10(9)/L vs.11.66 x 10(9)/L, P=0.013), hemoglobin level [(61.45 +/- 13.46) g/L vs. (75.89 +/- 23.11) g/L, P=0.003] and serum lactate dehydrogenase [(621.47 +/- 335.85) U/L vs.(1566.64 +/- 1720.45) U/L, P=0.020], while no differences were found between two groups in age, gender ratio, initial platelet count, percentage of blast in bone marrow, immunophenotypes and the expression of myeloid antigen CD13, CD33 and CD34. The prednisone sensitivity test showed that all 12 TEL-AML1 positive patients were good responders, while there were 11 prednisone poor responders among 40 negative patients (27.50%, P < 0.05). Bone marrow examination on day 15 showed no difference in the rate of complete remission between TEL-AML1 positive and negative patients.

CONCLUSIONThe incidence of TEL-AML1 fusion gene in cases of ALL is 21.05%. The load of leukemia cells in TEL-AML1 positive patients is significantly smaller than its counterparts, and the blast cells in TEL-AML1 positive patients are more sensitive to prednisone, indicating childhood ALL with TEL-AML1 fusion gene has a favorable prognosis.

Adolescent ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Gene Fusion ; Humans ; Infant ; Infant, Newborn ; Male ; Phenotype ; Platelet Count ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; genetics ; immunology ; pathology ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-ets ; genetics ; RNA ; isolation & purification ; Repressor Proteins ; genetics

Ginsenoside Rb3 (GRb3) is one of the main components in plasma of Panax quinquefolius Saponin of stem and leaf (PQS), which can be into human plasma. Previous studies have found PQS has estrogen-like vascular protective effects. In the present study, we investigated the estrogen-like protective effect of GRb3 on oxidative stress and dysfunction of endothelial cells induced by oxidized low-density lipoprotein. The activities of SOD, NOS and the contents of MDA in the cell lysate were examined by enzyme method or spectrophotometry. The NO and ET-1 concentrations in the cell culture supernatant were measured by ELISA method. The iNOS and eNOS mRNA expression were measured by real time RT-PCR, while the phosphorylation levels of Akt was measured by Western blotting. The results showed that GRb3 could enhance the activity of SOD, reduce the content of MDA, increase the level of NOS, NO, ET-1 and iNOS mRNA expression while decrease the eNOS mRNA expression and the phosphorylation level of Akt. These effects were blocked by estrogen receptor antagonist ICI182780. GRb3 can play a role in protecting vascular endothelial cells by estrogen receptors, the protective mechanism is similar to 17-β estrodiol.
Cells, Cultured ; Endothelial Cells ; drug effects ; Endothelin-1 ; metabolism ; Estradiol ; analogs & derivatives ; Estrogens ; pharmacology ; Ginsenosides ; pharmacology ; Humans ; Lipoproteins, LDL ; adverse effects ; Nitric Oxide Synthase Type II ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxidative Stress ; Panax ; chemistry ; Phosphorylation ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the role of tissue factor (TF) expression in the invasive and metastatic ability of colorectal carcinoma and explore the influence of TF on the invasive ability of HT-29 cells.

METHODSTF expression of specimens from 85 colorectal carcinomas and 6 colorectal adenomas was observed by immunohistochemistry. The role of TF expression in prognosis and tumor invasion and metastasis was analyzed. The plasmids pcDNA3.1/Zeo bearing either sense or antisense-TFcDNA were transfected into HT-29 cells by the way of Lipofectamine 2000. TF proteins in transfected and untransfected HT-29cells were detected by Western blot. In vitro Matrigel invasion assays were performed to show the invasive ability of those cells.

RESULTSTF expression was positive in 40 (47.1%) of 85 colorectal carcinoma specimens, but negative in normal mucosa and adenoma specimens. TF expression showed significant correlation with tumor invasive depth (r = 0.895, P < 0.01). TF expression showed significant correlation with synchronous and metachronous hepatic metastasis (r = 0.974, P < 0.01 and r = 0.963, P < 0.01 respectively). TF expression was a significant risk factor for hepatic metastasis (P < 0.01) and prognosis (P < 0.01). TF expression in HT-29 cells with sense/antisense-TFcDNA transfection was more/less than that of the cells without transfection. The invasive ability of HT-29 cells with sense-TFcDNA transfection was increased in vitro compared with the untransfected cells, but HT-29 cells with antisense-TFcDNA transfection got the contrary change.

CONCLUSIONSTF may take part in the invasive and metastatic process of primary colorectal carcinoma, and TF expression may be an indicator of hepatic metastasis and prognosis for colorectal carcinoma patients. TF expression may increase the invasive ability of HT-29 cell in vitro.

Blotting, Western ; Cell Movement ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; HT29 Cells ; Humans ; Immunohistochemistry ; Logistic Models ; Multivariate Analysis ; Thromboplastin ; analysis ; genetics

OBJECTIVETo evaluate the effect of combined resection for the treatment of T(4) gastric cancer and to refine the indication for en bloc dissection.

METHODSClinical data of 69 cases receiving combined resection were analyzed retrospectively, and compared with those of 45 cases undergoing palliative gastrectomy.

RESULTSOf 69 cases, 54 patients underwent curative combined resection, 15 patients underwent palliative combined resection, including 24 with transverse colectomy, 22 with pancreatico-splenectomy, 8 with left lateral lobectomy of liver, 6 with pancreatico-splenectomy and transverse colectomy, 5 with pancreaticoduodenectomy, 2 with cholecystectomy, 1 with splenectomy, 1 with phrenectomy. The total rate of lymph node metastasis was 88.4%; the operative mortality rate was 4.3%; the morbidity rate was 14.5%. The postoperatively 1-, 3-, 5-year survival rates of CR group and NCR group were 66.9%, 39.1%, 26.8% and 33.4%, 7.4%, 0% respectively (P < 0.01). The five year survival rate of curative resection group was 34.1%.

CONCLUSIONSAn en bloc combined resection can cure some T(4) patients, and improve the five-year survival rate.

Adult ; Aged ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Pancreatectomy ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery ; Survival Analysis ; Treatment Outcome

OBJECTIVETo investigate the clinical diagnosis, treatment and immunohistochemical characteristics of rectal stromal tumors.

METHODSImmunohistochemical expression of CD117 was retrieved in 20 cases of mesenchymal tumors of the rectum. And we analyzed the immunohistochemical characteristics, clinical data of rectal stromal tumors, and the results retrospectively.

RESULTSSixteen cases of rectal stromal tumors, 3 cases of leiomyosarcomas and 1 case of schwannoma were diagnosed. Histologically, 2 cases of GISTs were classified as benign, 1 as borderline and 13 as malignant. All of rectal stromal tumors (100%) were strongly positive for CD117, and 14 cases (88%) positive for CD34. The demographic profile of rectal stromal tumors showed a male predominance with average age of 60 years old. The main symptoms were urinary retention, constipation and abdominal pain. 14 cases were positive in digital rectal examination. The recurrence rate of local and radical resection in malignant stromal tumors was 4/4 and 3/6. 1, 3, 5-year survival rates were 89%, 64%, 48%, respectively. After operation mean survival time of was 47 months.

CONCLUSIONSThe specific GIST constituted the majority of mesenchymal tumors in rectum. It usually showed malignant biological behavior. Invasion and recurrence were common. Earlier diagnosis and radical resection had better prognosis. Periodically following up can help to detect the recurrence timely.

Adult ; Aged ; Female ; Gastrointestinal Stromal Tumors ; diagnosis ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Rectal Neoplasms ; diagnosis ; pathology ; surgery ; Retrospective Studies