OBJECTIVETo investigate the pro-apoptotic effect of Her-2 targeted recombinant caspase-6 fusion protein on osteosarcoma SOSP-9607 cells.

METHODSRecombinant immunocasp-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody (e23sFv), a PEA translocation domain (PEA aa253-364) and an active caspase-6. The immunocasp-6 gene was cloned into pCMV plasmid to construct a kind of eukaryotic expression vector, i.e. pCMV-e23sfv-PE II-caspase-6 (abbr. pCMV-6) and transfected into SOSP-9607 cells. Murine xenograft models were randomly divided into two groups that received i.m. injections of liposome encapsulated pCMV-6 or pCMV alone. The tumor volume and weight of the nude mice and the tumor weight of the cured mice were observed and statistically analyzed. The morphological changes of the tumors were examined with HE staining, apoptotic morphology of the tumor was observed by TUNEL staining and the gene expression was analyzed by immunohistochemical staining.

RESULTSThe tumor growth of the mice in the treatment group was significantly slower than that of the control group (P = 0.001). The weight of the nude mice in the treatment group was significantly higher than that of the control group (P = 0.0002). The tumor weight of the mice in the treatment group was significantly lower than that of the control group (P = 0.0006). HE and TUNEL staining of the tumor of nude mice in the treatment groups showed typical characteristics of apoptosis, while normal structure was found in the control group. Furthermore, caspase-6 was not found in the tumor and muscle tissues in the control group, but only in the treatment group by immunohistochemistry.

CONCLUSIONImmunocasp-6 can selectively recognize and bind to and kill HER-2 positive osteosarcoma cells, therefore, to offer some foundation for the clinical treatment of osteosarcoma.

ADP Ribose Transferases ; genetics ; Animals ; Apoptosis ; Bacterial Toxins ; genetics ; Bone Neoplasms ; metabolism ; pathology ; Caspase 6 ; genetics ; metabolism ; Cell Line, Tumor ; Exotoxins ; genetics ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Random Allocation ; Receptor, ErbB-2 ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden ; Virulence Factors ; genetics

OBJECTIVETo analyze the outcomes of pregnancies in women with systemic lupus erythematosus (SLE) and the risk factors affecting the outcomes.

METHODSThe data of SLE patients with pregnancy admitted from October, 2006 and September, 2015 were analyzed for assessing the maternal and fetal outcomes and complications. Their risk factors affecting the outcomes of the pregnancies were analyzed.

RESULTSThe 66 SLE patients (69 pregnancies) had a mean age at SLE diagnosis of 22.9 ∓ 5.1 years with a mean duration of SLE of 4.1∓3.6 years before pregnancy. Forty-five (65.2%) of the patients received oral medication for SLE treatment during pregnancy, and 44 (63.8%) were treated with prednisone and 19 (27.5%) were treated with hydroxychloroquine. The highest SLEDAI score was 6.8∓7.4 during pregnancy. The patients with moderate-to-severe disease activity had a higher rate of fetal loss (12 [54.5%] vs 12 [25.5%]) with a significantly lower birth weight of the newborns than those with remittent or mild disease (2073.0∓ 778.7 vs 2817.8∓533.7 g, P<0.05). The patients with moderate-to-severe disease activity also had higher rates of new-onset SLE (9 [40.9%] vs 6 [12.8%]), hypertension (12 [54.5%] vs 3 [6.4%]), active lupus nephritis (22 [100%] vs 4 [8.5%]), pneumonia (5 [22.7%] vs 2 [4.3%]), and renal insufficiency (8 [36.4%] vs 2 [4.3%]) compared with patients with remittent and mild disease (P<0.05). Active lupus nephritis (OR=6.10,95%CI: 1.43-25.96) was a significant predictor of adverse outcomes of the pregnancies.

CONCLUSIONFetal loss and maternal complications are common in patients with SLE in relation with the disease activity. Active lupus nephritis is a predictor for poor outcomes of pregnancies in SLE patients.

Marfan syndrome is a systemic disorder of connective tissue, caused by mutations in the FBN1, TGFBR1 or TGFBR2 genes. This syndrome is characterized by involvement of three major systems, skeletal, ocular, and cardiovascular. The continuing improvements in molecular biology and increasing availability of molecular diagnosis in clinical practice allow recognition of Marfan syndrome in patients with incomplete phenotypes. Additionally, molecular analyses could also be used for preimplantation genetic diagnosis. The identification of a mutation allows for early diagnosis, prognosis, genetic counseling, preventive management of carriers and reassurance for unaffected relatives. The importance of knowing in advance the location of the putative family mutation is highlighted by its straightforward application to prenatal and postnatal screening.
Fibrillin-1 ; Fibrillins ; Humans ; Marfan Syndrome ; diagnosis ; genetics ; pathology ; Microfilament Proteins ; genetics ; Mutation ; Prenatal Diagnosis ; ethics ; methods ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, Transforming Growth Factor beta ; genetics

Animals ; Chronic Disease ; Febuxostat ; Heart Failure ; drug therapy ; metabolism ; Humans ; Reactive Oxygen Species ; Thiazoles ; pharmacology ; therapeutic use ; Xanthine Oxidase ; antagonists & inhibitors

OBJECTIVETo evaluate the outcome of in vitro fertilization embryo transfer (IVF-ET) in treatment of polycystic ovarian syndrome (PCOS) with infertility.

METHODSA retrospective analysis was performed from 52 patients with PCOS (PCOS group) and 408 cases with tubal infertility (control group). Both groups underwent IVF-ET treatment from 2001 to 2004. The duration of stimulation, amps of gammaFSH, the level of serum E2, P on the day of HCG injection, the number of oocytes retrieved, the rates of fertilization, cleavage, implantation and pregnancy, the incidence of ovarian hyperstimulation syndrome (OHSS) and cancelled rate of ET were compared between the two groups.

RESULTThe duration of stimulation and amps of gammaFSH were not significantly different between the two groups. The concentration of serum E2, P on the day of HCG injection, the numbers of oocytes retrieved and cleavaged embryos were significantly higher in PCOS group (P <0.01, <0.05). Fertilization rate was significantly lower in PCOS group (P <0.01). The implantation, pregnancy and miscarriage rates per ET were not statistically significant. The OHSS rates and cancelled rates of ET were higher in PCOS group (P <0.01).

CONCLUSIONWomen with PCOS have a lower fertilization rate compared with those with tubal-factor fertility during IVF-ET. However, more oocytes are recovered and the preimplanted embryo has a normal chance of implantation leading to similar pregnancy rates. The OHSS rates and cancelled rates of ET are higher in PCOS because of a greater number of oocytes developed and a higher level of E2.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; etiology ; Ovarian Hyperstimulation Syndrome ; Polycystic Ovary Syndrome ; complications ; Retrospective Studies ; Treatment Outcome

BACKGROUNDTissue engineering techniques combined with gene therapy have been recently used to improve osteogenesis. NEL-like molecule-1 (Nell-1), a novel growth factor, has been reported to have specificity for osteochondral lineage. The study assessed the osteogenic differentiation of rat bone marrow stromal cells (bMSCs) after Nell-1 gene modification and examined its ectopic bone formation ability in a nude mice model with tissue engineering technique.

METHODSbMSCs obtained from Fischer 344 rats were transduced with either AdNell-1 (Nell-1 group) or Ad-beta-galactosidase (AdLacZ, LacZ group) or left untransduced (untransduced group). The expression of Nell-1 protein was determined by Western blotting and transfer efficiency was assessed. mRNA expressions of osteopontin (OP), bone sialoprotein (BSP) and osteocalcin (OC) were assessed by real-time PCR 0, 3, 7, 14, and 21 days after gene transfer. Alkaline phosphatase (ALP) activity was measured and von Kossa test was also conducted. Finally, with a tissue engineering technique, gene transduced bMSCs, combining with beta-tricalcium phosphate (beta-TCP) at a concentration of 2 x 10(7) cells/ml, were implanted at subcutaneous sites on the back of nude mice. Four weeks after surgery, the implants were evaluated with histological staining and computerized analysis of new bone formation.

RESULTSUnder current transduction conditions, gene transfer efficiency reached (57.9 +/- 6.8)%. Nell-1 protein was detected in Nell-1 group but not in untransduced group and LacZ group. Induced by Nell-1, BSP and OP expression were increased at intermediate stage and OC expression was increased at later stage. ALP activity and the number of calcium nodules were highest in Nell-1 group. Four weeks after implanted into nude mice subcutaneously, the percentage of new bone area in Nell-1 group was (18.1 +/- 5.0)%, significantly higher than those of untransduced group (11.3 +/- 3.2)% and LacZ group (12.3 +/- 3.1)% (P < 0.05).

CONCLUSIONSThis study has demonstrated the ability of Nell-1 to induce osteogenic differentiation of rat bMSCs in vitro and to enhance bone formation with a tissue engineering technique. The results suggest that Nell-1 may be a potential osteogenic gene to be used in bone tissue engineering.

Alkaline Phosphatase ; metabolism ; Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; metabolism ; ultrastructure ; Integrin-Binding Sialoprotein ; Male ; Mice ; Mice, Nude ; Microscopy, Electron, Scanning ; Nerve Tissue Proteins ; metabolism ; Osteocalcin ; genetics ; Osteogenesis ; Osteopontin ; genetics ; Rats ; Rats, Inbred F344 ; Reverse Transcriptase Polymerase Chain Reaction ; Sialoglycoproteins ; genetics ; Stromal Cells ; cytology ; metabolism ; ultrastructure ; Tissue Engineering

OBJECTIVETo investigate the effects of activated AKT on murine myeloid precursor cells (32D cells), and the effects of IFN-γ on 32D cells and its mechanisms.

METHODSPlasmid transduction was used to enhance the expression of AKT on 32D cells. After the transfected cells treated with IFN-γ for 24 hours, proliferation rate was tested by WST-1, apoptosis by flow cytometry, expression of phosphorylated Erk1/2, Stat3 and phosphorylated Stat3 was determined by Western blot.

RESULTS(1) IFN-γ at low concentration (100 U/ml) enhanced the growth and proliferation of 32D cells, while at high concentration (1000 U/ml) suppressed them. (2) Compared with control groups, low concentration IFN-γ increased (1124 ± 13) Stat3 phosphorylation in 32D-cell, while it high concentration IFN-γ decreased (601 ± 13). 32D cells transfected with activated Akt grew rapidly (0.287 ± 0.010) and had a low apoptotic rate [(9.57 ± 0.17)% (P < 0.05)]. (3) The expression of p-Erk1/2 in transfected 32D-cell was significantly reduced (P < 0.05). (4) Apoptosis rate of IFN-γ treated group was significantly decreased in transfected 32D cells (P < 0.05).

CONCLUSIONSIFN-γ has dual effects on 32D cells, namely, at low concentration enhanced the growth and proliferation of 32D cells, while at high concentration suppressed them. Its mechanisims is possibly through Stat3 pathway. Activated Akt can significantly promote the growth and proliferation of 32D cell and significantly inhibit apoptosis and IFN-γ can regulate cell proliferation and apoptosis through AKT. AKT activation can inhibit the Erk signal pathway, which may be affected by inhibition the modificaton of Raf1.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Phosphorylation ; drug effects ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; drug effects

BACKGROUNDIntraoperative magnetic resonance imaging (iMRI) dates back to the 1990s and has been successfully applied in neurosurgery but they were low-field iMRI (< 1.0T). This paper reports the clinical experience with a 3T iMRI-integrated neurosurgical suite in Huashan Hospital, Shanghai, China.

METHODSFrom September 2010 through March 2012, 373 consecutive patients underwent neurological surgery under guidance with 3T iMRI. A retrospective analysis was conducted regarding clinical efficiency.

RESULTSAll surgery in the 373 patients was safe. The ratio of gross total resection for cerebral gliomas (n = 161) was increased from 55.90% to 87.58%. The ratio of benefit in extent of resection was 39.13%. One hundred and fifty eight of the 161 glioma patients accomplished follow-up at 3 months postoperatively. Twenty of 161 patients (12.42%) suffered from early motor deficit after surgery. Late motor deficit was however observed in five of 158 patients (3.16%). Twenty-one of 161 patients (13.04%) had early speech deficit and late speech deficit was only observed in six of 158 patients (3.80%). The ratio of gross total resection for pituitary adenomas (n = 49) was increased from 77.55% to 85.71%. The ratio of benefit in extent of resection was 10.2%. There were no iMRI-related adverse events even for patients who underwent awake craniotomy.

CONCLUSIONThe 3T iMRI integrated neurosurgical suite provides high-quality intraoperative structural and functional imaging for real-time tumor resection control and accurate functional preservation, resulting in an improvement in maximal safe brain surgery.

Adolescent ; Adult ; Aged ; Brain Neoplasms ; surgery ; Child ; Child, Preschool ; China ; Female ; Glioma ; surgery ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Young Adult

Objective:Our study was aimed to analyze the therapeutic effect of early sequential enteral nutrition on postoperative rehabilitation in patients with gastric cancer.Methods:Patients with gastric cancer receiving surgery at our hospital from 2016 to 2017 included and the clinical information was prospective collected and analyzed.Patients were randomly divided into two groups using random number table.Patients in group A were sequentially given amino acid type,short peptide type and then whole protein type,while those in group B received whole protein formulation only.The recovery of gastrointestinal function,postoperative systemic inflammatory response,six-minutes walking test,and enteral nutrition-related complications were compared between the two groups.Results:A total of 71 patients were included in this study (Group A 36 cases,Group B 35 cases).There was no significant difference in terms of the restart anal exhaust between the two groups (P ＞ 0.05).Patients in group A had a significantly shorter postoperative hospitalization (t =4.070;P ＜ 0.01) and the earlier restoration of oral intake than that of Group B (t =3.400;P =0.001).One week after surgery,the levels of CRP (t =2.547;P =0.013) and IL-6 (t =3.172;P =0.002) were significant lower in group A when compared with group B.In addition,patients in group A had a significant higher six minutes walk steps than those in Group B [(416.1 + 36.7) m vs (358.9 ± 32.7) m;t =6.927,P ＜ 0.01].However,no significant difference in enteral nutrition-related complications was found between the two groups (P ＞ 0.05).Conclusion:In patients with gastric cancer,early sequential enteral nutrition can effectively accelerate the postoperative rehabilitation.

OBJECTIVE: To analyze the influence of preoperative serum nutritional indexes and postoperative nutritional guidance on 1-year mortality in elderly patients with hip fracture. METHODS: From January 2015 to December 2017, 396 elderly patients with hip fracture were included in the study, including 267 females and 129 males, aged 68 to 80(75.48±2.62) years; the course of disease was 2 to 10 (6.12±1.35) days;all patients were followed up for 1-year, and were divided into death group and survival group according to whether the patients died or not. Multivariate logistic regression model was used to analyze the influencing factors of 1 year mortality. RESULTS: Duringthe follow-up, 4 patients lost contact and were treated as shedding, among which 67 patients died and 325 patients survived. The age, male patients, patients with more than three basic diseases, American Society of Anesthesiologists grade Ⅲ-Ⅳ and patients with postoperative complications in the death group were significantly higher than those in the survival group (all CONCLUSION Advanced age, male and multiple underlying diseases were independent risk factors for 1-year mortality in elderly patients with hip fracture, while higher preoperative nutritional level and routine nutritional guidance were protective factors.
Aged ; Female ; Hip Fractures/surgery* ; Humans ; Logistic Models ; Male ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Risk Factors