Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Camptothecin ; administration & dosage ; analogs & derivatives ; Chemotherapy, Adjuvant ; methods ; trends ; Colorectal Neoplasms ; pathology ; Fluorouracil ; administration & dosage ; Humans ; Liver Neoplasms ; drug therapy ; secondary ; surgery ; Organoplatinum Compounds ; administration & dosage ; Preoperative Care ; Survival Analysis

The assisted reproductive techniques in China have witnessed rapid development in the past two decades.However,it also brings up issues of ethics and laws,especially the urgent regulation and reinforcement of the procedure of informed consent.This article begins with an explanation of the procedure of informed consent.Then it discusses the decision of the content,major and difficult points of informed consent in assisted reproductive technology services based on the practice in this field.Finally,it discusses the challenges to informed consent.To well enforce an informed consent process,we should first ensure the contents are reasonable and just;second,clinicians should have a scientific understanding of the importance of informed consent and establish normative procedure of informed consent;third,we must work according to relative ethic demands.Last but not least,respecting patients is the key to the principle of ethics.

Objective To investigate the clinical, electrophysiological and pathological features of critical illness polyneuropathy and myopathy (CIPNM). Methods The clinical outcomes, electromyogram Results as well as pathological features in nerves and muscles of 3 patients with CIPNM were investigated and analyzed. Results 3 patients were all provided with assisted respiration after tracheal intubation. 7～10 d after intubation, all the patients emerged muscle strength and tendon reflexes of extremities weakening; while 14 days after that, 2 patients appeared amyotrophy of extremities. Electromyogram showed that the conduction of many motor and sensory nerves for extremities decreased, while the amplitude of compound muscle action potential (CMAP) of part of motor nerves decreased. Biopsy for nerves revealed decreased medullated nerve fibers and regeneration phenomenon of auxiliary fibers; while that for muscles showed neuralgic damage and myopathy-like changes. Conclusion CIPNM can complicate after tracheal intubation. The electrophysiological and pathological examinations for nerves and muscles can be helpful for the diagnosis.

Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

Human herpesvirus 7 (HHV-7) infection is dependent on the functions of structural glycoproteins at multiple stages of the viral life cycle. These proteins mediate the initial attachment and fusion events that occur between the viral envelope and a host cell membrane, as well as cell to cell spread of the virus. To characterize the HHV-7 glycoproteins that can mediate cell fusion, a cell-based fusion assay was used. 293T cells expressing the HHV-7 glycoproteins of interest along with a luciferase reporter gene under the control of the T7 promoter were cocultivated with SupT1 cells transfected with T7 RNA polymerase. HHV-7 glycoproteins gB, gH, gL and gO can mediate the fusion of 293T cells with SupT1 cells, and the fusion can be inhibited by anti-CD4 mAbs. Thus, the coexpression of HHV-7 gB, gO, gH and gL is sufficient and necessary for HHV-7 induced membrane fusion, and one of these glycoproteins or protein complex formed by these glycoproteins might be the ligand(s) of CD4 molecule.

OBJECTIVETo explore the clinical outcomes of percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP) in treating osteoporotic vertebral compression fracture (OVCF).

METHODSFrom January 2007 to February 2010, the data of 40 patients with osteoporotic vertebral compression fracture underwent treatment were retrospectively analyzed. Of them,20 patients were treated with PVP (PVP group), there were 8 males and 12 females with an average age of (66.37 +/- 2.34) years old (54 to 81); 20 patients were treated with PKP (PKP group), there were 11 males and 9 females with an average of (65.12 +/- 3.21) years old (56 to 79). Postoperative at 1 week, 12 weeks, 1 year, pain and daily life function were respectively assessed by visual analogue scale (VAS) and Barthel index (BI); and anterior height of responsibility vertebra, Cobb angle were measured by X-rays.

RESULTSIn PVP group, 1 case complicated with bone cement leakage without clinical symptoms and no operation to treat. No postoperative infection and deep vein thrombosis were found between two groups. All patients were followed up more than 1 year, pain and daily life function has obviously improved than preoperative (P < 0.01); and there was no significant difference on 1 week, 12 weeks, 1 year after operation (P > 0.05); there was no significant difference between two groups (P > 0.05). In PVP group, there was no significant difference in anterior height of responsibility vertebra, Cobb angle before and after operation;and in PKP group, postoperative data has obviously improved than preoperative (P < 0.01), but there was no significant difference postoperative at 1 week, 12 weeks, 1 year (P > 0.05); there was no significant difference between two groups at 1 week, 12 weeks, 1 year after operation.

CONCLUSIONBoth the methods can obviously relieve pain and completely or partly recover daily life function in treating OVCF. But PKP has advantages of recovery of anterior height of responsibility vertebra and correction of Cobb angle, especially for serious compression.

Aged ; Aged, 80 and over ; Female ; Fractures, Compression ; diagnostic imaging ; physiopathology ; surgery ; Humans ; Kyphoplasty ; Male ; Middle Aged ; Osteoporotic Fractures ; diagnostic imaging ; physiopathology ; surgery ; Radiography ; Recovery of Function ; Retrospective Studies ; Spinal Fractures ; diagnostic imaging ; physiopathology ; surgery ; Spine ; surgery ; Treatment Outcome

AIM: To clone EBV-LMP2A gene, construct and identify the recombinant retroviral vector and stable cell strains expressing EBV LMP2A. METHODS: The full-length EBV LMP2A gene was generated by RT-PCR amplification from B95.8 cells which contain complement nucleotide sequence of EBV LMP2A gene. The gene was ligated to T-vector and sequenced to construct retroviral vector consisting with LMP2A. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pGEZ-LMP2A and two auxiliary viral vectors pHIT456 and pHIT60 by lipofectAMINE2000. Viral titration was performed according to the instructions of the manufacturer. To establish L929 cell line stable expressing LMP2A, L929 cells were infected with recombinant retrovirus three times and selected by Zeocine. The Zeocine-resistant clones (L929/LMP2A) were screened for LMP2A expression by RT-PCR and Western blot. RESULTS: The recombinant retrovirus vector carrying LMP2A gene was constructed successfully. Transfection yield a titer of 5×10~8 infectious particles/L. The infected L929 cells were selected by Zeocine. Results of RT-PCR and Western blot indicated that L929 transgenetic cells could stably express EBV-LMP2A. CONCLUSION: The L929 cell line stably expressing LMP2A provides suitability for extraction of the LMP2A protein and preparations of the vaccine for the therapy of EBV-associated diseases.

ObjectiveTo study the effect of α7 ( α7 AChR) agonist nicotine on regulating sensitivity of regular chemotherapeutic agent in cholangiocarcinoma cells,and explore the possible target.MethodsThe effect of nicotine and α-BTX pretreatment on the survival ability of cholangiocarcinoma cells was investigated when applied with 5-FU by using MTT and Flat cloning formation experiment.ResultsApplied with 5-FU,in various con centrations nicotine stimulating group( 10-3 g/L,10-4 g/L,10-5 g/L ),the survive rate of QBC939 was 128％,124％,118％,while that in α-BTX stimulating group and combined stimulation group was 92％,94％,93％,92％,respectively.The cloning formation ability of nicotine- stimulating group (6.2 ± 0.40) was significantly higher than α- BTX stimulating group (3.2 ± 0.20 ),combined stimulation group ( 3.2 ± 0.20 ) and control group ( 3.4 ±0.33).ConclusionNicotine can prevent chemotherapy-induced apoptosis,and improve cholangiocarcinoma cell survival via α7 nicotine acetylcholine receptor in vitro.

Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

To investigate the antitumor activities of the immunoconjugates composed of anti-type IV collagenase monoclonal antibody Fab' fragment and lidamycin (LDM) prepared with different linkers. The immunoconjugates were prepared by linking Fab' to lysine-69 of LDM apoprotein by SPDP, LCSPDP, SMBS or SSMPB as the intermediate drug linkers. Immunoreactivities of the conjugates were determined by ELISA. The cytotoxicities of the conjugates were examined by clonogenic assay. In vivo antitumor effects of the conjugates were evaluated in nude mice bearing subcutaneously implanted HT-1080 tumor. ELISA assay showed that the conjugates retained part of the immunoreactivity of 3G11 against the antigen. The cytotoxicities of the Fab'-SMBS-LDM and Fab'-SSMPB-LDM to HT-1080 cells were significantly potent, compared with Fab'-SPDP-LDM, Fab'-LCSPDP-LDM and free LDM. In animal models at the same condition, free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM inhibited the growth of HT-1080 tumor by 70.9%, 74.8% and 72.3%, while Fab'-SMBS-LDM and Fab'-SSMPB-LDM reached 78.0% and 87.7%, respectively. The median survival time of the mice treated with free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM were prolonged by 71.9%, 82.2% and 107.5%, respectively, compared with that of untreated group. Whereas, the median survival time of Fab'-SMBS-LDM and Fab'-SSMPB-LDM were prolonged by 145.2% and 165.8%, respectively, indicating that Fab'-SSMPB-LDM was more effective than Fab'-SMBS-LDM in tumor suppression and life span prolongation. Fab'-SSMPB-LDM has more marked selective antitumor efficacy and lower toxicity, and might be a novel candidate for cancer therapy.