Abstract:Objective. To study thymus-dependent T cell development and T cell repertoire in human s ever com-bined immunodeficiencypatients after HLA-identical or haploid entical T cell-depleted allogeneic bone marrow transplantation.Methods .Blood samples were obtained from15SCID patients before transplantation and a t varying intervals thereafter.Quantitative competitive PCR assay and immunosco pe analysis of the T cell receptorVarepertoire were performed.Results. Before and within the first100days after transplantation,patients’ periphera l blood mononuclear cellpresented an oligoclonal or polyclonal skewed T cell repertoire,low T cell re-ceptor excision circlesvalues and pred ominance of CD45RO + T cell.In contrast,the presence of high numbers of CD45RA + T cells in bone marrowcirculation reconstituted SCID patients(>10 0days post-transplantation)correlated with active T cell production by the th ymus as revealed by high TREC values,and a polyclonal T cell repertoire demonst rated by a Gaussian distribution of Va-specific peaks.Conclusions.Within one year after BMT ,a normal T cell repertoire develops in SCID patients as a resu lt of thymic output.The T cell receptor diversity is highly and positively corr elated in these patients with TREC levels.
Abstract:Objective.To develop monoclonal antibodies again st the catalytic subunit of human telomerase reverse transcriptasefor i ts expression detection of human tumors.Methods.A dominant epitope in hTERT was automatically synthesized based on Fmoc method,and was used to immuni ze Balb/c mice.Hybridomas were generated and screened by ELISA for specific mo noclonal antibodies,and the characterization was performed by Western blotting and immunohistochemical staining.The heavy chain variable region of antibody wa s cloned by RT-PCR and sequenced.Results.Antigenic peptide hTERT 7 was synthesized and confirmed by MALDI-TOF-MS and HPLC analysis.One hybr idoma cell line secreting anti-hTERT 7 antibodies designated as M2was established after primary screening and cons equent 3rounds of limited dilution.M2was IgG1in isotyping.The competi-tive assay showed that the M2antibody was hTERT 7-specific,and the affinity constant was about 1×10 6 mol-1 .The antibody reacted with cell extracts from HeLa cancer cells but not wi th those from normal2BS cells in ELISA assay.For in situ staining of immunohis tochemistry,the positive staining presented in the nuclear compartment of HeLa ,while2BS was negative.The heavy chain variable region from M2re-vealed tha t the monoclonal antibody was mouse origin.Conclusions.The developed mouse mon oclonal antibody is hTERT-specific and able to recognize native cellular hTERT in ELISA and immunohistochemistry,which makes the immuno-detection of telom-e rase hTERT expression in cancer cells or tissues possible.
Abstract:Objective.To evaluate the effects of high-fructose diet on expression of orex in and its receptors,orexin1receptor and orexin2receptor in rat hypothalamus tissue,and to analysis the interaction of related fa ctors involved in regulating orexin and its receptors.Methods.Insulin resistan ce rat model induced by high fructose confirmed by the gold standard eug-lycaem ic clamping was employed and mRNA expression of orexin and its receptors OX 1 R and OX 2 R in hypothalamus,mRNA expression of leptin in adipose tissue were measured by reverse transcription poly-merase chain reaction.Serum insulin and triglyc eride levels were measured by chemiluminescence im-munoassay and biochemical en zyme techniques.Results.Expression of orexin mRNA decreased about 40%in hi gh fructose diet rats compared to control group,whereas expressi on of orexin1receptor and orexin2receptor mRNA increased up to4.4and5.1f old.Leptin mRNA expression in adipose tissue increased about 3 0%in comparison with control group.Blood glucose,serum insuli n and triglyceride have shown signi ?ficant higher levels than those in contro l group.Glucose infusion rate was much low-er in comparison with control group.Conclusi ons.High?fructose diet induces insulin resistance in rats with impact on orexi n and leptin regulations.Blood glucose,serum insulin,lipid metabolism and lep tin play an interactive role on orexin and its receptors regulation in rats.
Keywords:orexin;dietary fructose;insulin resistan ce
Abstract:Objective.To study expression and localiza tion of Smad4protein,the common-mediator Smad,which is one of intracellular signaling molecules of transforming growth factor-afamily,in rat t estis during postnatal development.Methods.In this study,whole testes were co llected from S.D rats aged3days,7days,14days and28days,and adult respe ctively.We examined the cellular localization and developmental change of Smad 4in rat testis by immunohistochemical ABC method with glucose oxidase-DAB-nic kel enhancement technique;the quantitative analysis of the immunostaining by t he image analysis system;the Smads pro-teins coexistence in the adult rat test is by the double immune staining for CD14-Smad4and Smad2-Smad4;the prote in expression of Smad during rat testicular development by means of Western blo ts.Results.The protein of Smad4was present in rats from3days of age to adul thood,and the im-munolocalization was exclusively localized to the cytoplasm o f Leydig cells with negative nuclei in the in-terstitial tissue at any time po int.No expression was detected in germ cells.The result of image and sta-tis tical analysis showed that generally,there was a tendency that the expression o f Smad4in the testes increased gradually with the rats developing maturation.C onclusion.Our data provide direct evidence for the molecular mechanism of TGF-aaction in rat testes during postnatal development and spermatogenesis of ra ts.
Keywords:tran sf orming growth factor;Smad4;testis
Abstract:Objective.To d etermine whether the non-expression of DPC4protein only occurs late in the dev el-opment of colorectal carcinoma.Methods.In this study,we examined the ex pression of DPC4protein in formalin-fixed archival specimens from102colorec tal neoplasm with immunohistochemical analysis.Those specimens were classi-fie d into5stages:stageⅠ;stageⅡ(intramucosal carcinoma ,8cases);stageⅢ(primary invasive carcinoma without infiltration of the l ymph nodes,11cases);stageⅣ(primary invasive carcinoma with infiltration of the lymph nodes,25cases);and stageⅤ(carcinoma metastasized to dis-ta nt tissue,22cases).Results.The frequency of non-expression of DPC4prote ins were5.5%in stage I,12.5%in stage II;9% in stage III;36%in stage IV;32%in stag e V.The frequency of negative expression of DPC4protein were analyzed by÷ 2 test for stage II and III versus stage IV and V and there was statistically significant difference.At same time ,there was statistically signifi-cant differencefor adenoma versus carcinoma .Conclusions.The frequency of non-expression of DPC4protein increases as the stage of colorectal carcinoma advances and the non-expressio n of DPC4protein is likely to be a late event in the sequential pathogenesis o f colorectal carcinoma.The non-expression DPC4protein in colorectal neoplasm may sug-gest its malignant characteristic,which will help us to increase the insight on colorectal carcinoma.
Abstract:Objective.To in vestigate clinicopathologic characteristics and surgical results of nonfunctiona l islet cell tumors.Methods.We performed retrospective analysis of50patient s with nonfunctional islet cell tumor treat-ed at Peking Union Medical College Hospital from July1968to July1999,and summarized clinical symptoms and signs,primary diagnosis before surgery,surgical treatments,pathologic and im munohisto-logical characteristics.Results.Of the50cases examined from July 1968to July1999,12were found during physical examinations. The most common symptoms were upper abdominal pain and upper abdominal dis-com fort,which appeared in20casesand17casesrespectively,an d the most common sign was abdominal masses.B-ultrasound and CT are the most commonly used means for preoperative examination,with posit ive rates of97.8%and100%respectively.Op-e rative mortality was2%.Five-year survival rate of radical surgery of maligna nt cases was75%.Immuno-histological examinations showed that tumors rich i n multi-peptide linked hormones,neuron-specific eno-lase and chromaffin were in65%cases,90.6%and73.3% respectively.The positive rates of insulin,glucagons,somatostatin,pancreat ic polypeptide,gastrin and vasoactive intestinal peptide were68.2% ,51.2%,42.9%,40% ,25%and15.8%respectively.Conclusion.Nonfuncti onal islet cell tumors lack diagnostic specificity.Ultrasound and CT are primar y examination methods.Immunohistological analysis indicates different hormones ,but lacks the presence of related clinical symptoms.Surgery is an effective treatment for nonfunctional islet cell tumors,and even for those with distal m etastasis,immediate surgical removal and treatment can improve prognosis.
Abstract:Objective.To describe the effects of surgical treatment a nd complications in46patients with carotid body tumor.Method.Retrospe ctive study on surgical treatment and complications was carried out in46case s of CBT which were surgically treated with different kind of procedures.Resul t.All procedures performed successfully except that the CBT was not excised in 4cases.No operative mortality was observed.There were2postoperative hemiple gia,4hypoglossal nerve impairment ,2glossopharyngeal nerve impairment ,1va gus nerve impairment ,and1accessory nerve impairment.One patient presented p ostoperative cranial nerve impairment in glossopharyngeal,vagus and hypoglossa l nerves.Two patients developed local recurrence during the long-term follow-up.Conclusion.Complete surgical excision was possible in each patient if th e diagnosis had been cor-rectly made through selective preoperative angiography ,vessel ultrasound Doppler and other examinations.Reasonable surgical procedur e and Matas training were necessary to the successful surgical treatment and t hus decrease the complicative incidence rate of carotid chemodectomas.
Keywords:carotid body tumor;surgical tr ea tment;complication
Abstract:Objective.T o assess the viability of surgical procedures on gynecologic malignant patients o f70years age and older.Methods.Between September1,1983to June30,1 999,57gynecologic malignant patients aged70years and older were treated by surgical procedures.A retrospective study was perf ormed.All patients were analyzed for preexisting medical conditions,length of hospital stay,morbidi-ty,and mortality.Results.Thirty-four patients had a n extensive surgical procedure,while a local surgical procedure was done in23 patients.Forty-one patientshad one or more preexisting medical i llnesses.Minor surgical morbidity occurred in24patientsand majo r surgical morbidity occurred in6patients.There were no differen ces in the types of surgical procedures,mean hospital stay,preexisting medical illness and postoperative complications between the two groups of patients70 to75year older and over age75.The major postoperative complications all occ urred in the extensive surgical procedure group that were higher as compared wit h local operation and postoperative mean stay was also significant-ly length in former group.Conclusions.The extensive surgical procedure can be performed fo r elderly patients with gynecologic malignances.Careful preoperative evaluatio n,monitoring,and meticulous postoperative care are vital to the success.
Abstract:Objective.To c ompare the analgesic efficacy and adverse effects of patient-controlled analges iawith tramadol and with morphine for postoperative middle or severe pain .Methods.Fifty-nine patients,scheduled for elective hysterectomy or hystero myomectomy ,were ran-domly divided into Group Tand Group M.The2drugs were administered intravenously v ia a patient-controlled analgesia device till24h postoperatively.Efficacy wa s assessed by comparing total pain reliefand the sum of pain intensity differencevalues over24h.Results.Statistically significant equival ence of tramadol and morphine was shown by TOTPAR values(15.9±4.4and1 6.4±3.5,respectively)and SPID values(9.2±4.7and9.0±2.0, respectively).Tramadol caused fewer adverse events than morphin e(16.7%and26.7%of patients,respectively).Conclusion.The analgesic efficacy of PCA with tramadol and with morphine were equivalent in the treatme nt of postoperative pain,and tramadol can cause slighter gastrointestinal adve rse effects.
Abstract:Objective.To test whether intramuscular,intranasal,intrarectal and intravagina l administration of HPV6b L 1 virus-like particlescould induce immune response in mice and to as sess whether intra-muscular and mucosal vaccination against HPV is feasible.Me thods.HPV6b L1proteins self-assembled into VLPs in Sf-9cell in vitro.Mic e were immunized on day0and21with50ìg HPV6b L1VLPs intramuscularly,int ranasally,intrarectally and intravagi-nally respectively.Sera were collected for testing IgG titer after a further7days and3months respec-tively.Results .After immunizations,all mice developed significant anti-HPV6b L1antibody titers in serum by7days after the second immunization.The titer of the serum I gG antibody against HPV6b L1VLPs in the intramuscularly immunized group was h igher than that in the intranasally,intrarectally and intravaginally immunized groups respectively,indicating that both muscular and mucosal administration of HPV6b L1VLPs can stimulate a systemic HPV-specific antibody response.Sera of the mice in the in-tramuscularly immunized group still maintained a high tit er of the serum IgG antibody against HPV6b L1VLPs 3months after the immunizat ion.Conclusion.The results demonstrated that the HPV6b L1VLPs maintain stro ng antigenicity.Immu-nization with HPV6b L1VLPs via intramuscular and mucos al routes,without adjuvant ,can elicit spe-cific antibody in sera.These fin dings suggest that the VLPs are able to induce protective antibodies.
Abstract:Objective.To investigate the effects of Ginkgo biloba extracton cer ebral vasospasmafter subarachnoid hemorrhageand its influence on n itric oxideand endothelin-l .Methods.Noncraniotomy models of SAH in Wistar rats were used and animals were divided into sham-operated group ,SAH group,saline-treated group and EGb-treated group.Diameter of basilar a rtery was measured.Regional cerebral blood flow,NO and ET-1levels in blood,and calcium content in brain tissue within24hours after SAH were dete cted.Pathological examination of hippocampus CA1sub-field was also performed .Results.Sham operation did not alter the above parameters.Induction of SAH l ed to a marked spasm of basilar artery.rCBF decreased obviously and consecutive ly within24hours after SAH.Meanwhile NO level in serum decreased,ET-1level in plasma and calcium content in brain tissue significantly in-creased.Pyra midal cells in hippocampus CA1subfield were severely damaged.EGb significantly antago-nized the pathological alterations of the above parameters.Conclusion .Alterations of NO,and ET-1play an important role in the development of CV S after SAH.EGb exerts its protective effects on CVS by inhibitng the above pat hological alterations.
Abstract:Objective.To improve the clinical knowledge of lymphangiomatosis.Methods.We pre sented a case of a30?year?old woman where generalized lymphangiomatosis pre-sented with an inguinal mass.Multiple imaging modalities were undertaken,inc luding plain film,CT,MRI ,US,and lymphangiography.Splenectomy was performe d.Results.The bones,lungs,spleen and retroperitoneal lymphatics were involve d.The diagnosis was made by integration of clinical,imaging and pathological f indings.Also a literature review regarding clin-ical presentation,diagnosis, management and outcome of this rare disease was made.Conclusion.we suggest the importance of clinical knowledge of the entity and imaging procedures are es sential for better diagnosis,thus avoiding more invasive procedures.