Objective: To construct subtracted cDNA libraries of stomach tumors and normal stomach tissue using suppression subtractive hybridization(SSH).Methods: cDNA Library subtraction was performed using the protocol described in the Clontech PCR-Select cDNA Subtraction Kit. cDNA was synthesized from 2 μg of poly A+RNA from the tumor and normal tissues using AMV reverse transcriptase. The tester and driver cDNAs were digested with RsaⅠ, a four-base-cutting restriction enzyme that yields blunt ends. The tester cDNA was then subdivided into two portions, and each was ligated with different cDNA adaptor. Two hybridizations were performed. In the first, an excess of driver was added to each sample of tester. Hybridization kinetics led to equalization and enrichment of differentially expressed sequences. During the second hybridization, the two primary hybridization samples were mixed together without denaturing and thus the templates were generated from differentially expressed sequences for PCR amplification. Using suppression PCR, only differentially expressed sequences were amplified exponentially and after second PCR amplification the background was reduced and differentially expressed sequences were further enriched. The cDNAs were then directly inserted into a T/A cloning vector to generate a stomach tumor-specific subtracted cDNA library. Results: The amplified library contained 800 positive clones. Plasmid inserts were PCR amplified and showed 250－700 bp inserts. Conclusions: The successfully constructed subtracted cDNA library of gastrocarcinoma and normal tissue enables us to compare two populations of mRNA and obtain clones of genes that expressed in one population but not in the other.The characterization of these genes will allow them to be exploited for their diagnostic and therapeutic potentials.

Objective To compare the results of the three grades pictures affiliated With the Synotophore and to choose the opti- mal pictures for the clinician.Design Retrospective case series.Participants 41 normal persons and 44 cases of intermittent exotropia. Methods All participants were separately examined the three grades function with different kinds and degrees pictures affiliated with the Synotophore.The average radian measured with four sorts of pictures of the first grade function and the average measured with the three sorts of pictures of the second grade function were analysed,the stereoacuity of third grade function was measured with qualitative picture and random dot quantitative picture.Comparison was made between two groups of the people who possessed with stereoacuity. Main Outcome Measures Radians and seconds gained from the pictures affiliated with the Synotophone,Results Synotophore of the superposition of the first grade function:the outcome of using four sorts of pictures to examine the normal persons were 1.47+1.68, 1.21?1.21,1.68?1.60,1.32?1.27,F=0.595,P=0.619;the intermittent exotropias were 8.41?4.30,8.73?4.26,8.45?3.86,8.73?4.28,F= 0.070,P=0.987;the difference was not prominent.The fusion range of the second grade of Synotophore,using three different pictures to examine the normal persons were 26.03?6.17,21.11?6.87,17.21?5.40,F=17.260,P=0.000,the result of the fusion range has direct proportion with the degrees of the pictures,the bigger the picture was,the larger the fusion range,the difference was statistically signif- icant.And there was no significant differences for the intermittent exotropias when examined with the same pictures(21.74?7.43,21.31?7.82,20.32?7.53),F=0.590,P=0.556.The stereoaeuity of the third grade of Synotophore,in the normal person group 85.4% and 100%, X~2=4.877,P=0.027,the results were statistically significant between the qualitative picture and the random dot quantitative picture. About the intermittent exotropia group,no statistically significant differences were detected(72.73% and 59.10%),X~2=1.821,P=0.177. Condusion Choose pictures randomly when examine the first grade function of Synotophore;for the second grade function of Syno- tophore in both groups may choose intergraded degree pictures;the stereoacuity of the third grade of Synotophore,normal group should choose quantitative pictures,while strabismic patients may choose both of them.(Ophthalmol CHN,2008,17:120-122)

We aimed to study the effect of allitridum (All) on the transient outward potassium current (Ito) of ventricular myocytes of spontaneously hypertensive rats (SHR). Totally 30 male SHRs were randomly divided into three groups: low-dose All group (7.5 mg·kg(-1)), high-dose All group (15.0 mg·kg(-1)) and normal saline group. The other 10 sex and age matched Wistar-kyoto rats (WKY) were also taken as control group (WKY group). All animals received i.p. administration for 8 weeks. The dual enzymatic method was used to separate single ventricular myocyte from animals. Patch-clamp technique was used to record Ito and analyze the effect of All on the current. It was shown that the left ventricular hypertrophy of SHR was reversed significantly by All. Furthermore, the density of Ito was recovered in both high and low dose All groups. The peak current densities of Ito were enhanced from 18.23±3.64 to 25.17±2.86 pA/pF (P<0.01) and 36.47±5.42 pA/pF (P<0.01) at +50 mV by All 7.5 mg·kg(-1) and 15.0 mg·kg(-1), respectively, which was not significantly different with WKY group. The effect was associated with positive shift of the steady-state, close-state inactivation, and shortened recovery from inactivation of Ito. It is concluded that All decreases the remodeling of Ito of ventricular hypertrophic myocytes of SHR.
Allyl Compounds ; pharmacology ; Animals ; Hypertrophy, Left Ventricular ; drug therapy ; Male ; Myocytes, Cardiac ; cytology ; drug effects ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Sulfides ; pharmacology

The present study was carried out to determine the role of histamine H(1) and H(2) receptors in the generation of basic respiratory rhythm. Neonatal (aged 0-3 d) Sprague-Dawley rats of either sex were used. The medulla oblongata slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O(2) and 5% CO(2)), and ended in 3 min. Respiratory rhythmical discharge activity (RRDA) of the rootlets of hypoglossal nerve was recorded by suction electrode. Thirty medulla oblongata slice preparations were divided into 5 groups. In groups I, II and III, histamine (5 μmol/L), H(1) receptor specific antagonist pyrilamine (10 μmol/L) and H(2) receptor specific antagonist cimetidine (5 μmol/L) was added into the perfusion solution for 15 min separately. In group IV, after application of histamine for 15 min, additional pyrilamine was added into the perfusion for another 15 min. In group V, after application of histamine for 15 min, additional cimetidine was added into the perfusion for another 15 min. The discharges of the roots of hypoglossal nerve were recorded. Signals were amplified and band-pass filtered (100-3.3 kHz). Data were sampled (1-10 kHz) and stored in the computer via BL-420 biological signal processing system. Our results showed that histamine significantly decreased the respiratory cycle (RC) and expiratory time (TE), but changes of integral amplitude (IA) and inspiratory time (TI) were not statistically significant. Pyrilamine induced significant increases in RC and TE, but changes of TI and IA were not statistically significant. Cimetidine had no effects on RC, TE, TI and IA of RRDA. The effect of histamine on the respiratory rhythm was reversed by additional application of pyrilamine but not cimetidine. Taken together, with the results mentioned above, histamine H(1) receptors but not H(2) receptors may play an important role in the modulation of RRDA in the medulla oblongata slice preparation of neonatal rats.
Animals ; Animals, Newborn ; Cimetidine ; pharmacology ; Female ; Histamine ; pharmacology ; Histamine H1 Antagonists ; pharmacology ; Histamine H2 Antagonists ; pharmacology ; Hypoglossal Nerve ; physiology ; In Vitro Techniques ; Male ; Medulla Oblongata ; physiology ; Pyrilamine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Histamine H1 ; physiology ; Receptors, Histamine H2 ; physiology ; Respiration

Aged ; CD5 Antigens ; metabolism ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 14 ; Cyclin D1 ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Lymphoma, B-Cell ; metabolism ; pathology ; Lymphoma, Follicular ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; genetics ; metabolism ; pathology ; surgery ; Pseudolymphoma ; metabolism ; pathology ; Translocation, Genetic

Objective:To assess the feasibility and accuracy of real-time three dimensional echocardiography(RT-3DE) in quantitative evaluation of left ventricular(LV) volume and ejection fraction(EF) in patients with ventricular aneurysm and myocardial infarction.Methods: Twenty-three patients with left ventricular aneurysm combined with myocardial infarction were examined by RT-3DE,two dimensional echocardiography Simpson's method,and M-mode Teichholz method separately.The following parameters: left ventricular end diastolic volume(LVEDV),end systolic volume(ESV),stroke volume(SV) and EF were obtained by each method and the results were compared with those obtained by left ventriculography(LVG).Results: The values of LVEDV,LVESV,SV,and EF determined by RT-3DE showed good correlations with those determined by LVG(r=0.92,0.90,0.88,and 0.91,respectively;P(0.05).) The values of LVEDV,LVESV,SV,and EF determined by Simpson's method also showed good correlations with those determined by LVG(r=0.85,0.87,0.86,0.91,respectively;P

Objective To explore the modulatory effect of 5-HT2A receptors on the discharge activities of inspiratory neurons in medial region of nucleus retrofacialis of neonatal rats. Methods Experiments were performed in vitro brainstem slice preparations from neonatal rats. These preparations included the medial region of nucleus retrofacialis with the hypoglossal nerve rootlets retained. The rhythmic discharges of the inspiratory neurons and activities of the hypoglossal nerve rootlets were simultaneously recorded by using microelectrodes and suction electrodes, respectively. Roles of 5-HT2A receptors in modulation of the discharge activities of inspiratory neurons were investigated by administration of the 5-HT2A receptor agonist 1-(2,5-dimethoxy-4-iodopbenyl)-2-aminopropane (DOI), and its specific antagonist ketanserine dissolved in modified Kreb's solution for perfused slices. Results In DOI group, the inspiratory time (TI) was (0.864±0.07)s, expiratory time (TE) was (10.78±1.06)s, respiratory cycle (RC) was (11.79±1.64)s, integral amplitude (IA) was (357.98±37.21)(μV·s) and the peak discharge frequency (PF) was (37.83±3.66)Hz. In control group, they were (0.68±0.06)s, (13.89±2.14)s, (14.77±1.92)s, (273.57±24.39)(μV·s), and (29.92±4.50)Hz, there were significant differences between the 2 groups (Pa<0.01). In ketanserine group, TI was (0.55±0.07)s, TE and RC were (18.43±3.28)s and (20.17+2.91)s respectively, IA and PF were (214.37±33.52)(μV·s) and (22.17±3.92)Hz, there were significant differences between ketanserine group and DOI, control group (Pa<0.01). Conclusion 5-HT2A receptors take part in modulate the discharge activities of inspiratory neurons in neonatal rat brainstem slices.

Objective To explore the modulation roles of gamma-aminobutyric acid A(GABAA) receptor on respiratory rhythmical discharge activity in brainstem slice of neonatal rats.Methods Experiments were performed on in vitro neonatal rats brainstem slices.The slices containing the medial region of the nucleus retrofacialis(mNRF)with the hypoglossal nerve rootlets retained was prepared in which the respira-tory-related rhythmic discharge activity(RRDA)were recorded from hypoglossal nerve rootlets by suction electrode.The possible role of GABAA receptor on RRDA was investigated by administration of GABA and its antagonist bicuculline in modified Kreb′s solution(MKS),for perfusion of the brainstem slices.Eighteen slices positive of RRDA were randomly divided into three equal groups:the first group of slices was perfused with different concentrations(10,20,40,60 ?mol/L) GABA and the most effective concentration was selected;the second group of slices was perfused with 10 ?mol/L bicuculline;the third group of slices was perfused with 10 ?mol/L bicuculline plus 40 ?mol/L GABA.The RRDA in 3 groups were observed.Results GABA decreased RRDA,40 ?mol/L was the most effective concentration.It shortened inspiratory time(TI),decreased integral amplitude(IA),prolonged respiratory cycle(RC) and expiratory time(TE) respectively.By contrast,10 ?mol/L bicuculline produced opposite effects on RRDA,it prolonged TI,increased IA,shortened RC and TE respectively.40 ?mol/L GABA plus 10 ?mol/L bicuculline had no significant effects on RRDA.Conclusion GABA plays an important role in modulate RRDA of neonatal rats via GABAA receptor.

ObjectiveTo analyze the relationship between clinical characteristic pathology and prognosis in infant with intra-abdominal solid tumor.MethodsFifty-two infants(less than 1 year old) with abdominal solid tumor from Apr.1998 to Feb.2007 in Shanghai Children's Medical Center and from Jan.2001 to Feb.2007 in Shanghai Xinhua Hospital were reviewed.The history of these children were reviewed.Features and clinical pathology of these children with their prognosis were analyzed and followed up by telephones and children return visit records from 5 months to 8 years.ResultsLess than 1 month,7 cases;1 month to 1 year old,45 cases.Teratoma 23 cases(44.23%),neuroblastoma 9 cases(17.31%),nephroblastoma 6 cases(11.54%),hepatoblastoma 5 cases(9.62%),epithelioid hemangioendothelioma of the liver 3 cases(5.77%),congenital mesoblastic nephroma 3 cases(5.77%),fusiform cell epithelioid hemangioendothelioma of pancreas 1 case(1.92%),hamartoma of the liver 1 case(1.92%),retroperitoneal small cell malignant tumor 1 case(1.92%).Benign:malignant=1:1.Among the benign tumor,male:female=1:1.Among the malignant tumor,male:female=2.33:1.0.All children were treated with tumor resection,and combined with chemotherapy for those whose tumors were malignant.ConclusionsAmong infant abdominal solid tumors,teratoma and neuroblastoma are much more than other tumors.The cases of benign tumors are almost as much as the malignant tumors.The benign tumors did not have sex differences,and had good prognosis after surgical resection.However,in malignant tumors,the incidence rate of male is obviously higher than female.Completely resection of those malignant tumors with chemotherapy would get little incidence of recrudescence and low case fatality rate.Early diagnosis and early treatment play an important role in prognosis.

Objective To analyze plague epidemic tendency in the Three-River Region of Qinghai.Methods Using retrospective study,the Three-River Region during 1954-2006 year pestis epidemic focus were investigated and analyzed.Result Pestis prevailed mainly in Yushu,Chindu,Qumalai,Nangqian,Zhiduo and the Geermu.Tanghla Township.It was first found that the nature plague focus of miefitus existed in Chengduo County.There are 1 5 kinds of 12 branches in 8 trees infected plague animals were founded,336 Yersinia pestis were separated from the driven objects.Among them there were 291 Himalayas marmot body,account for 86.60%of the total,13 of Tibet sheep,accounts for 3.87%.10 of Qinghai field-mouse,accounts for 2.98%,Also there were 114 Yersinia pestis which were separated from each kind of vector insect in vivo.And,46 pestis strains came from the axe shape of flea in vivo account for 40.35%(46/114),38 pestis strains separated from Xie mountain flea,account for 33.33% (38/114).During 1960-2006 years there were 85 human plague cases were founded,238 occurred,134 died,the case fatality rate wero 56.30%(134/238),the popular seasons were started from May to November,the peak season happened in Aug and Sep.After Oct mainly due to Tibet sheep pestis which will cause as the origin of infection.The majority of sickness was pulmonary plague,account for 49.58%(117/238),whereas the first round case caused by the gland bubonic plague,account for 77.12%(91/118).Conclusions There are two pestis strains natural epidemic focus places in Three-River Source Region of Qinghai including the Himalayas marmot pestis strain and the Qinghai field-mouse pestis strain.The case of human pestis strain causes by the marmot strain,the fiehl-mouse mold mushroom spawn causes human pestis strain has not yet discovered,Three-River Source Region of Qinghai is a pestis strain key popular area in Qinghai Province.