Objective To construct the secretory expression vector of recombinant human C-reactive protein(rhCRP) for its se-cretory expression in Pichia pastoris ,rhCRP was expressed as a secretory protein and purified ,and the immunity reactivity of the purified protein was identified .Methods The DNA fragment of rhCRP which was designed and synthesized was cloned into pPICZαA vector .Recombinant plasmid pPICZαA/rhCRP was linearized by SacⅠand transformed into Pichia pastoris X-33 by elec-trotransformation .The rhCRP was secreted into the medium under the methanol induction .RhCRP was purified by Histamine affin-ity chromatography .The purified rhCRP was identified by SDS-PAGE and Western blotting ,and its immunity reactivity and stabili-ty was identified by indirect ELISA .Results The pPICZαA/rhCRP expression vector was successfully constructed .The rhCRP of 23 × 103 was inducted and successfully expressed as a secretory protein by the recombinant Pichia pastoris strains .The rhCRP was purified by one step up to 90 .42% purity ,and it was showed good immunity and stability by indirect ELISA .Conclusion The rh-CRP with higher purity and immunoreactivity was successfully obtained by using the Pichia pastoris expression system ,which pro-vided an important experimental basis for producing anti-human CRP antibodies and developing testing CRP reagent .

Objective To analyse the clinical features of pelvic insufficiency fractures (PIF) in patients with cervical cancer after radiotherapy and explore the risk factors affecting its occurrence .Methods 55 cases of patients with cervical cancer that used radia-tion therapy as a single treatment were chosen in our hospital from February 2007 to May 2010 and the clinical features were ana-lysed;The fractures of spine ,sacrum ,sacroiliac joint ,ilium and pubis and femoral were observed and the density of fifth lumbar (L5 ) vertebrae ,left and right sacrum were detected via the CT or MRI (Hounsfield Unit)after 2-year follow-up ,the different factors on the incidence of PIF were explored .Results 12 cases (19 sites) of patients in 55 patients occured PIF after follow-up 2years ,the cumulative incidence was 21 .82% ;the distribution of PIF parts were followed :lumbar vertebrae had 5 cases ,sacral had 5 cases ,pu-bis had 4 cases ,sacroiliac joint had 2 cases ,iliac had 2 cases and femoral head had 1 case .The incidence rate of PIF that density of fifth lumbar vertebrae was ≤130 HU and >130 HU were 16 .36% and 5 .45% respectively ,the PIF′incidence rate that density of left side of sacrum was≤ -35 HU and > -35 HU were 18 .18% and 3 .64% respectively ,the incidence rate of PIF that density of right side of sacrum was ≤ -35 HU and > -35 HU were 20 .00% and 1 .82% respectively ,the density of the three parts all had statistical difference on the incidence of PIF (P<0 .05) .Univariate analysis showed that menopausal status and the average density of CT had statistical effects on PIF (P<0 .05);Multiple analysis showed that the average density of CT was the independent factors affecting PIF′incidence after radiotherapy .Conclusion The patients with cervical cancer may occur pelvic insufficiency fracture af-ter radiotherapy ,and its incidence is related to menopausal status and bone mineral density .

Obejctive To compare the differences between conventional radiation therapy (3DCRT) and intensity modulated radiation therapy( IMRT) for locally advanced laryngeal carcinoma patients by ECLIPSE treat -ment planning system.Methods Six locally advanced laryngeal carcinoma patients ′treatment plans were de-signed as IMRT and 3DCRT according to ECLIPSE treatment planning system ,respectively.The following values were measured,such as V95%、D5%、D95%、Dmin and D mean of the target volume,and the D50,D33,D5, Dmax and Dmean of normal tissues.Results The dose coverage rate of GTV and GTVnd in the IMRT plan was compared to ECLIPSE treatment planning system with no statistical differences.However, there were obvious difference in CTV and some normal tissues.Conclusion Both IMRT and 3DCRT have good coverage to primary tumor and metastatic lymph nodes.IMRT can spare the parotid from high dose irradiation.

OBJECTIVETo evaluate the applicability of immune-related response criteria (irRC) in treating non-small cell lung cancer (NSCLC) by Chinese medicine (CM).

METHODSTotally 97 stage III a-IV NSCLC patients were predominantly treated with comprehensive CM. Curative effects were evaluated by three methods such as Response Evaluation Criteria in Solid Tumors (RECIST), Oncologic Curative Effect Evaluation Criteria of Chinese Medicine in Solid Tumor (draft, abbreviated as CM criteria), and irRC. The correspondency and consistency between irRC, RECIST and CM criteria were analyzed and compared. The objectivity of irRC in evaluating curative effect of Chinese medical treatment for NSCLC was assessed.

RESULTSThe correspondency rate of irRC to RECIST was 59. 79% with Kappa value of 0. 379 (U test, P <0. 01). The two criteria had certain correspondence, but with an unsatisfactory consistency. The correspondency rate of irRC to CM criteria rate was 83. 51% with Kappa value of 0.751 (U test, P <0. 01). The two criteria had good correspondence and consistency.

CONCLUSIONSCM criteria had good consistency with CM criteria in evaluating curative effect for Chinese medical treatment of advanced NSCLC. Its results could objectively reflect features and advantages of CM for treating advanced NSCLC.

Asian Continental Ancestry Group ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; immunology ; Medicine, Chinese Traditional ; standards ; Treatment Outcome

Objective To investigate the serum amyloid A (SAA) and interleukin-18(IL-18)concentration in the pathogenesis of type 2 diabetes mellitus (T2DM) and its macrovascular complications, and study the relation between them. Methods ELISA was used to assay serum SAA and IL-18 levels in 65 T2DM patients (including 31 cases with macrovascular complications) and 30 healthy controls. Results Serum SAA and IL-18 levels [(3.09±0.96)mg/L, (98.8±36.4)ng/L]were significantly elevated in patients with T2DM as compared with those in control subjects [(1.06±0.45)mg/L, (58.9±15.6)ng/L](P<0.05). There was significant difference of SAA and IL-18 levels between T2DM patients with [(6.34±1.52) mg/L,(141.2±48.3)ng/L]and without macrovascukar complications [(2.65±0.39)mg/L, (80.2±20.1)ng/L](P < 0.05).Univariate linear regression analysis showed significant positive correlations between serum IL-18 with SAA (r =0.615, P<0.05), SAA, IL-18 and fasting blood glucose (FBG) had mutual positive correlations (r=0.312, 0.428, P< 0.05, respectively). Conclusions In patients with T2DM, serum SAA and IL-18 concentration is greater than in non-diabetic subjects. SAA and IL-18 play important roles in the initiation and development of T2DM. The study suggests that SAA and IL-18 might be an important independent risk factor.

Objective To investigate the effects and signal transduction pathway of vasonatrin peptide(VNP),a novel man-made natriuretic peptide,on hepatic fibrosis.Methods Mice were injec ted with carbon tetrachloride(CCl4)for 12 weeks,with or without VNP treatment in the last 6 weeks.HE staining and Sirius red staining were performed to evaluate the status of hepatic fibrosis.In vitro after treatment of VNP,and DNA and collagen synthesis of cultured HSC-T6 hepatic stellate cells were assessed by[3H]-thymidine and[3H]proline incorporation,respectively.The signaling pathway involved was identified by radioimmunoassay to detect the levels of intracellular cGMP,and by mimicking experiments using 8-br-cGMP(a membrane-permeable cGMP analog).Blocking experi ments were performed using HS-142-1,an antagonist of guanylyl cyclase-coupled natriuretic peptide receptor(NPR),or KT-5823,the cGMP-dependent protein kinase(PKG)inhibitor.Results VNP markedly alleviated CCl4-induced liver fibrosis in mice.In vitro,HSC-T6 cells demonstrated a dosedependent reduction of DNA and collagen synthesis in the presence of VNP.In addition,VNP significantly increased intracellular levels of cGMP.The effects of VNP were mimicked by 8-br-cGMP,but they were inhibited by HS-142-1,or KT-5823.Conclusion VNP ameliorated liver fibrosis by inhibiting collagen production from hepatic stellate cells via guanylyl cyclase-coupled NPR/cGMP/PKG signal pathway,indicating that VNP might be a new effective agent in the treatment of liver fibrosis.

ObjectiveTo examine the relationship between HIF-1αt and VEGF expression and the clinicopathological characteristics in hypopharyngeal squamous cell carcinoma. Methods The expression status of HIF-1α and VEGF were examined by immunohistochemistric method (IHC) in 62 tumor tissue and 19 paracancerous normal tissue.The relationships between the expression of HIF-1α and VEGF and clinicopathological characteristic were analyzed. Results HIF-1αt and VEGF expression were higher in hypopharyngeal carcinoma tissues than those in paracancerous normal tissues (66. 1％:26. 3％ ;x2 =18. 02,P ＜0. 05 ;67. 7％ : 31.6％ ; x2 =19.22, P ＜ 0. 05 ). The expression intensity of HIF-1 α was related to T stage, N stage and TNM stage ( x2 =4. 23,5.83,9.94,all P ＜0. 05). The expression intensity of VEGF was related to metastasis, T stage, N stage and TNM stage (x2 =5.62,7. 38,15.75,4. 29 ,all P ＜0. 05 ). There was minus relationship between overall survival and expression level of HIF-1 α and/or VEGF (x2 =29. 25, P＜0.01; x2 =24.88, P＜ 0.01 ).On multivariate analysis,HIF-1α expression and T stage were independent prognostic factors for overall survival (x2 =4.80,5.74, all P＜0. 05).ConclusionsHIF-1α and VEGF may be considered as a parameter in evaluation of progression, metastasis and prognosis of hypopharyngeal carcinoma and also may be a direction of molecular target therapy.

Objective To evaluate the efficacy and safety of levetiracetam (LEV) monotherapy on children with epilepsy.Methods Forty-one children (26 cases were male,15 cases were female) with epilepsy aged 7 months to 13 years were treated with LEV as monotherapy.These patients were selected from Department of Neurology ,Wuhan Children′s Hospital, from Aug.2007 to Aug.2009.The starting do-sage of LEV was (13.6?4.7) mg?kg-1?d-1,twice daily,and its objective dosage was (25.7?7.5) mg?kg-1?d-1,twice daily.LEV monotherapy was investigated by a self-controlled and open-label research,and the follow-up period ranged from 6 months to 2 years.Results The effective rate was 68.3% (28 cases),with 39.0% (16 cases) achieving seizure freedom in LEV monotherapy of children with epilepsy.Thirteen patients (31.7%) had poor efficacy in reduction of seizures,7 patients (17.1%) discontinued LEV monotherapy due to either an inadequate seizure control or aggravated seizures.Fifteen patients (36.6%) had the therapy-related adverse events in LEV monotherapy,including gastrointestinal dysfunction (5 cases),irritability (5 cases),dizziness (2 cases) and somnolence (2 cases).The adverse effects appeared in 2-4 weeks of early LEV therapy and were spontaneously disappeared in 1 week to 1 month of continuing therapy.Conclusions The LEV monotherapy is effective and safe for the control of partial and generalized seizures in children with epilepsy.LEV appears to be a broad-spectrum,first-line anti-epileptic drug in treatment of children with epilepsy.

Objective To investigate the clinical characteristics and diagnostic main points of poliomyelitis-like syndrome(PS).Met-hods The clinical data of 15 children with PS were analyzed retrospectively including nervous system manifestation,results of lab andauxiliary examination,diagnosis and reason of misdiagnosis,treatment and progress.Results PS in all 15 children was characterized by mean age of onset(3.3 years),and forerunner infection(in 11 cases,73%)with acute upper respiratory infection of diarrhea 1-2 weeks pre-onset or during onset.All cases were acute flaccid paralysis,12 cases(80%) of which were only one limb.The positive outcome of serologic examination in PS consisted of 4 cases in coxsackie virus-IgM,1 case in EB virus-IgM,1 case in herpes simplex virus-IgM and 1 case in mycoplasma-IgM.All children showed the electromyologram changes in nerve damage.The muscle force of 4 cases increased one grade.Conclusions Children with PS are characterized by the age of onset under 5 years old,acute flaccid paralysis(mostly affected one limb),and the most pathogen being enterovirus.The electromyologram examination can help establish a definite diagnosis in PS.

Objective To analyze negative lymph nodes of 34 non-small cell lung cancer(NCLC) patients with total correction by means of fluorescent quantitation PCR and immunohistcchemistry,and to form molecular bi- ology staging.Methods Clinical data and tissue samples of 193 lymph nodes were collected from 34 patients under- going resection for non-small cell lung cancer.Using fluorescent quantitation reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry method,lymph nodes were examined for CEA gene mRNA,P53 and CK to form molecular biology staging.All the patients were followed-up for an average of forty months.Results The CEAmRNA was identified in 21.7% (42/193) lymph nodes negative patients from 17 patients(17/34,50%); TMN staging was up-regulated in 8 patients;positive lymph nodes were increased in 9 patients.P53 and AE1/AE3 were identified 9.8%(19/193) from 11 patients,18.6 % (36/193)from 15 patients,separately;TMN staging was up-regulated in 2 patients of P53 examination and 5 patients of AE1/AE3 analysis;positive lymph nodes were in- creased in in 7 patients of P53 examination and 11 patients of AE1/AE3 analysis.There was obvious statistical sig- nificance in them,but the molecular biology staging based on the three markers was not an independent factor on re- currence and metasis of lung cancer.Conclusion CEAmRNA.P53 and AE1/AE3 analysis could find lung cancer micrometasis more sensitively to form molecular biology staging which was relative to the prognosis,but not an inde- pendent prognostic indicator.It might be good to the therapy strategy after operation.