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Volume 0 ， Issue 2 ， 1998
- Abstract：The present study reports an improved PCR-based technique that allows quick and effective screening of cDNA libraries. First, the cDNA library was arrayed as follow: about 3 X 10’ cDNA clones were multiplied as individual plaques on solid medium in 24-well culture dishes at 1 200 plaque forming units per well. The phage suspension of each well was transferred to an individual microcentrifuge tube in 72-tube box. Then, box pool, row pools and column pools were set up that respectively represent a 72-tube box, rows and columns within the box. To screen a specific target cDNA,primers specific for novel ESTs ob- tained in our laboratory were employed to conduct PCR in a hierarchy mode. PCR began with the box pools, resulting in the identification of some Positive box pools. Then PCR went down to the row and col- umn pools of the positive box. The intersection of the positive row （s） and column （s） revealed the candi- date positive tubes. The specificity of PCR products were meanwhile checked by restriction enzyme diges- tion. Finally, hybridization was carried out to get single specific cDNA clomes from the positive tubes. This PCR-based technique features high specificity, high efficiency and less-cost in large-scale cDNA library screening. Our initial implementation of the technique resulted in the isolation of three longer different cD- NA clones from a human fetal brain cDNA library. Thus this improved technique can serve as an alterna-tive to the time-consuming and laborious conventional hybridization-based method for screening cDNA li-brary.Keywords：arrayed cDNA library;hierarchy PCR;hybridization4|5|6Updated：2024-04-10
- Abstract：Orphanin FQ（OFQ） or nociceptin is a novel neuropeptide consisting of 17 amino acids. This peptide has a primary structure reminiscent of that of opioid peptide but exhibits an opposite effect to make animals hyperreactive. The effect of this new peptide on cardiovascular function are not completely known. The present study was conducted to investigate the effect of intravenous bolus injection of orphanin FQ on mean arterial blood presure （MABP） in conscious stroke-prone spontaneously hypertensive rats (SHRsp). Adult male SHRsp and Wistar normotensive rats （250～300 g body weight, 2. 5～3 months old） were used in this study. The MABP was measured in the conscious state by a tail-cuff method. In SHRsp model, intravenous bolus injection of orphanin FQ or Tyr1-orphanin FQ （0. 5 mg/kg） induced a prolonged and marked reduc- tion in MABP. The maximum changes in MABP were -30. 2±4. 2 mmHg by orphanin FQ and -28. 2± 4. 7 mmHg by Tyr1-orphanin FQ at 10 min after administration,and this effect lasted over 30 min. The Phe1→Tyr substitution in orphanin FQ was found to retain almost fully hypotensive activity. Pretreatment of SHRsp with naloxone-HCI（60 μg/kg）, 5 min before the injection of orphanin FQ, did not block the hy- potensive effect of orphanin FQ. Therefore, opioid receptors could not account for the hypotensive effect of orphanin FQ in SHRsp. In Wistar rats, intravenous bolus injection of the same dose of orphanin FQ did not cause a change in MABP. These observations suggest that orphanin FQ is a novel hypotensive peptide and may have some role in the regulation of blood pressure in SHRsp, rather than in normotensive rats. The ex-act underlying mechanisms are waiting to be clarified.Keywords：orphanin FQ;naloxone 1 stroke-prone spontaneously hypertensive rats (SHRsp)4|15|2Updated：2024-04-10
- Abstract：Objective. To investigate whether the polymorphisms in the angiotensin converting enzyme （ACE） gene and angiotensinogen （AGT） gene are associated with essential hypertension. Methods. A case-control study was carried out using 103 hypertensive （HT） and 131 normotensive （NT） subjects. The insertion/deletion（I / D ） polymorphism of the ACE gene and the methionine→threo- nine variant at position 235 （M235T） of the AGT gene were determined by the polymerase chain reaction （PCR） technique and PCR/restriction fragment length polymorphism （PCR/RFLP） analysis, respective- ly. Results. The differences of D allele frequency and genotype distribution of the ACE gene between NT and HT groups were statistically significant （X2=18.12, P<0. 005 ）. The T235 allele frequency of the AGT gene was 69% in NT Chinese group （approximately 1. 38 to l. 64 fold that in Caucasians）, and was greater in female HT than in NT （0. 82 vs 0. 72, X2= 8. l, P<0. 025）. A correlation between M235T molecular variant of the AGT gene and I/D molecular variant of ACE gene to hypertension was found. Cbeclusions. The possession of D allele of the ACE gene might be a marker for predisposition to hyper- tension. The T235 allele of the AGT gene was more common in Chinese than in Caucasians, and might contribute to the risk for hypertension in female Chinese.Keywords：essential hypertension;renin angiotensin system;gene polymorphism4|30|7Updated：2024-04-10
- Abstract：Expression vector inserted with E2/NS1 gene derived from genotype Ⅲ Chinese isolates of HCV was transfected into mammalian cells to express E2 glycoprotein. Expressed protein was used as antigen for an- ti-E2 antibody detection in 19 cases of hepatitis C patients by Western blot. It was first to express E2 gly- coprotein of genotype Ⅲ Chinese hepatitis C virus isolates. For anti-E2 detection, 14 cases of patients were positive of antibodies against E2（73. 7 % ）. These results indicated that E2 glycoprotein expressed in mam-malian cells had good immunogenicity and cross reactivity to serum infected with genotype Ⅱ Chinese hep-atitis C virus isolates.Keywords：hepatitis C virus;protein expression;genotype4|5|0Updated：2024-04-10
- Abstract：Objective. Atopic asthma provides a useful model for evaluating the genetic factors that control human immune responsiveness. HLA class Ⅱ gene products are involved in the control of immune response. As HLA-DRB gene is the most polymorphic HLA class Ⅱ gene, we investigated whether susceptibility or resistance to the disease is associated with HLA-DRB. Methods. Blood samples were obtained from two groups of unrelated Chinese northern adults: （l） 50 atopic asthma （7 of them with familial aggregation） ;（2） 80 healthy controls without asthma or atopy and other HLA-associated diseases. Genomic DNA was extracted from peripheral venous blood leucocytes. The polymorphic second exon of HLA-DRB gene was amplified by sequence-specific primer polymerase chain reaction （SSP/PCR） methods. All patients had their serum IgE （total and specific） antibody levels by RAST, bronchial reactivity assessed by methacholine brocho-provocation test and/or bronchodilation test. Results. There was an increased gene frequency of DR6（13） and DR52 in asthmatic subjects compared with healthy subjects（17% vs 4. 3%, P<0.01; 50% vs 17. 5 %, P<0. 01 ）, and the decreased frequency of DR2（15） and DR51 in asthmatic patients（7% vs 18%, P<0. 05; 2 % vs 33 %, P<0. 01 ）. We found the positive association between DR6（13）-DR52 and sIgE antibody responsiveness to dl （from house dust mite al- lergen ）; negative association between HLA-DRB alleles and TIgE or BHR （bronchial hyperresponsiveness ）. Conclusion. The results suggested that HLA haplotype DR6（13）-DR52 was significantly implicated in sus- ceptibility to house dust mite induced-asthma, at least it would be more closely associated with atopic asth- ma. Conversely, alleles DR2（15） and DR51 might confer protection against the disease. HLA-DRB genes were particularly involved in regulating human atopic immune response in asthma.Keywords：asthma;human leukocyte antigen4|8|2Updated：2024-04-10
- Abstract：Objectival. To prepare a radioimmunoguided surgery （RIGS） system including radio-nuclides hand- held gamma detecting probe （GDP） with signal processing control unit, specific monoclonal antibody （MAb）, technique of labeling as well as detection of ovarian carcinoma in nude mice bearing human ovari-an carcinoma. Metals. MAb COCl83B2, produced in our laboratory was used to prepare the F （ah’）2, fragment by ficin digestion. 99mTc labeling of COC183 B2-F（ab’）2, and normal mice IgG were prepared according to im- proved pretining method. GDP and control unit were made by Chinese Atomic Energy Research Institute. Different groups of nude mice bearing human ovarian carcinoma with ascites were administered with differ- ent dosages of 99mTo COCl83 B2-F（ab’）2, and their tissues measured by GDP in vivo and compared with gamma well counts after dissection in vitro. Results. 99mTc labeled COCl83B2-F（ab’）2, yielded a labeling rate of over 90%, and a good immunoac- tivity. The detections of carcinoma tissue by GDP showed a T/NT ratio of 2: 1 as positive for cancer. The smallest carcinoma detected was 0. 2cm × 0.1 cm and the nearest distance between carcinoma and normal tissue was 0. 2cm. Conclusions. The above studies proved that ficin can be used to digest MAbs with high yields and good immunoactivity of F（ab’）2. Our RIGS system with 99mTo labeled COC183 B2-F（ab’）2 and GDP can hope-fully be used clinically for ovarian carcinoma detection.Keywords：ovarian carcinoma;monoclonal antibody F（ab’）2;radioimmunoguided surgery4|7|0Updated：2024-04-10
- Abstract：Objective. This study was to investigate the effects of transforming growth factor-β（TGFβ） and fi- broblast growth factor （FGF） in the subcapsular opacification formation of the lens. Methods. Lens epithelial explants from 10-day-old rats were cultured with TGFβ1 or TGFβ2 in the presence of FGF for 5 days, then were examined by light and electron microscopy, and by immunolocal- ization of smooth muscle（α-sm） actin and type I collagen. Results. In TGFβ/FGF-treated explants,extensive proliferation occured, with formation of spindle and star-shaped cells. These cells showed ultrastructure and biochemical features of fibroblast or myofibroblast. Prominent Golgi apparatus and rough endoplaic reticulum were observed in some cells. Intracellular micro- filaments with cytoplasmic dense babies and membrane associated dense bodies, features of smooth muscle cells, were also observed. Some cells showed reactivity to -sin actin antibody. TGFβ/FGF-treated ex- plants were strongly stained with type I collagen antibody. Condusion. In the presence of FGF, TGFβ1 and TGFβ2 induced lens epithelial cell （LEC ） proliferation and transformation into fibroblast or myofibroblast-like cells, with producing of abundant collagen matrix in the explants. The changes are similar to the metaplasia that occurrs in subcapsular opacification of the lens. The findings suggest that TGFβ and FGF plays a role in the pathogenesis of subcapsular opacification of the lens.Keywords：transforming growth factor-β;fibroblast growth factor;lens4|4|0Updated：2024-04-10
- Abstract：Breath-hold clue MRI was employed for the identification of severe coronary artery stenoses after 0. 56 mg/kg of dipyridamole was infused. Fourteen patients without myocardial infarction but with ≥70% di- ameter narrowing of 1 or 2 major coronary arteries were studied. Each patient underwent coronary angiog- raphy, MRI at rest and during stress. Segmental wall motion abnormalities were visually assessed in a clue loop, followed by quantitative analysis by calculation of percent systolic wall thickening（%SWth）. The re- suits showed that the sensitivities of 70% and 90% were present for the qualitative and quantitative analy- ses, respectively, in respect to the approximate specificities（82 % & 86% ）. Further quantitative analysis showed that sensitivities and specificities were 88% and 88% for l-vessel disease versus 92% and 83% for 2-vessel disease; 86 % and 100% for LAD, 100% and 70% for LCX, 89% and 100% for RCA. We con- cluded that quantitative analysis is significantly superior to qualitative analysis for the identification of se- vere coronary stenosis while d ipyridamole- induced wall motion abnormalities were assessed by breath - hold cine MRI.Keywords：dipyridamolel magnetic resonance imaging;ischemic heart disease4|9|0Updated：2024-04-10
- Abstract：The gonadal histopathology, and its correlation with the clinical features has been investigated in 9 true hermaphroditism patients, aged 5-21 yr. Seven of 9 patients had been raised as females, of which the chromosomal karyotype was 46XX in 5 cases, 46XX/46XY and 46XX/47XXY in 2 cases. Two of 9 pa- tients were raised as males, the chromosomal karyotypes being 46XX and 46XX/46XY. All 9 patients had testicular tissue excised, and biopsies of the conserved ovarian tissue were performed. Ovotestis was the most common form of the abnormal gonads; two of 9 patients had bilateral ovotestes, seven had unilateral ovotestes （5 in right side, 2 in left side）. In seven patients with a unilateral ovotestis, 6 had a contralateral ovary and one had a contralateral testis. Microscopically, the ovarian tissue of 11 ovotestes, including 6 biopsies from contealateral ovaris, were normal, with many primordal follicles and a few growing follicles. In two of the patients, aged over 15 years, evidence of ovulation was observed. In comparison, the testicu- lar tissue of the ovotestis and the one left inguinal testis was histologically abnormal, with immature semi- niferous tubules, most of which filled with Sertoli cells only. Three of 9 patients married after surgical treatment. Two of these subsequently conceived and delivered of normal infants by cesarean section.Keywords：true hermaphroditism;gonad;histopathology4|14|0Updated：2024-04-10
- Abstract：Objective. This study sought to investigate the effects of early and long-term intervention with an- giotensin-converting enzyme（ACE） inhibitor captopril on the elderly patients with acute myocardial in farc- tion（AMI）, and observe its in-hospital and post-hospital outcomes during serial follow-up of 54 months. Methods. 631 elderly patients（60～75 years old） with AMI and without cardiogenic shock were hospi- talized within 72 hours of symptoms and were randomly allocated to captopril （n = 361;treatment group） and conventional treatment （n = 270; control group）. The survival and cardiac events （congestive heart fail- ure, reinfarction, severe arrhythmias and cardiac death）of each group were determined during hospitaliza-tion and follow-up. Results. During hospitalization, the survival was higher in treatment group than in control group（p< 0. 0001 ）. On the other hand, in treatment group lower mortality was true for patients with anterior my- ocardial infarction（p < 0. 001 ） or with anterior+inferior myocardial infarction （P= 0. 026 ）, but not statis- tically significant in ones with inferior myocardial infarction （P= 0. 061 ）. During follow-up, the occurrence of cardiac death, heart failure, reinfarction and severe arrhythmias were lower in treatment group （P = 0. 0001, P = 0. 05, P = 0. 0004 and P = 0. 027）. So higher survival （P = 0. 005 ） and lower total cardiac events（p= 0. 0008） could be seen in treatment group over this period. Conclusions. Early and long-term treatment with captopril in the elderly patients with AMI has bene-ficial outcomes in both in-hospital and follow-up periods.Keywords：elderly;acute myocardial infarction;angiotensin-converting enzyme inhibitor4|7|0Updated：2024-04-10
- Abstract：After the rats were inflicted with 8 Gy total body gamma ray irradiation and 15 % total body surface area （TBSA） full thickness burn injury, they were treated with blood transfusion （BT） and bone marrow transplantation （BMT）. Then the survival of allografts grafted on the escharectomized burn wounds in the 24, 48 and 72 h postinjury was observed. It was found that when the burn wounds were closed with allo- grafts in the 24h postinjury, there were an early elevation of leucocytes, the appearance of the donor’s cells and a significantly higher survival rate of the rats on the 30 day postinjury. The allografts could survive longer and wounds showed no signs of infection and healed quicker. When the allografts were grafted in the 48 h or 72 h postinjury, only harmful effects to hasten the death of rats were observed.Keywords：combined radiation-burn injury;blood transfusion;allograft;BMT4|10|2Updated：2024-04-10
- Abstract：Sixteen patients with osteoarthritis （13 knees and 3 hips）, 3 patients with rheumatoid arthritis （RA） and 4 cadaver were studied for evidence of immune complex in the destroyed articular cartilage tissues. Frozen sections of the articular cartilage from arthroplasty were stained with fluoresceinated antibodies to human immunoglobulins IgG, IgA, IgM and complement C3. The results showed: 1. There were immune complexes linear deposited in the surface of the irregular articular cartilage tissues and on some chondro- cytes remained in most patients with osteoarthritis （14/16）. The patterns of immune complexes are IgA, complement C3, IgG and IgM, their percentage is 81. 25%, 75%, 75% and 50% respectively. 2. In all of 3 patients with RA, the surfaces of articular tissues were seen with patchy diffusely positive areas for IgA, IgG, IgM （excepting negative in I case） and complement C3. 3. There were no immune complexes deposited in the surfaces of 4 cases of normal articular tissues. The presence of immune complexes in the cartilages suggested that an autoimmune reaction participated in the pathological process of osteoarthritis and that the autoimmunity may be responsible for the continuous degeneration of the osteoarthritis.Keywords：autoimmune;pathology;osteoarthritis4|14|0Updated：2024-04-10
- Abstract：Five cases of arytenoid dislocation after intubation under general anesthesia are reported in this article. The anatomical and anesthetic causes of this complication were studied restrospectively, and the methods of treatment were discussed. The result showed that the incidence of arytenoid dislocation was about 0. 09% （5/5 826）. Hoarseness, dysphonia and difficulty in feeding were the prevalent symptoms, and poor mobili- ty of the vocal fold was the most common sign. The treatment included closed reduction of the displaced arytenoid under local anesthesia, steroid and physical therapy. The possible causes of arytenoid dislocation were summarized as follows: （1） trauma produced by the blade tip of laryngoscope, （2） the use of stylet, （3） overstretching of aryepiglottic fold and vocal cord during laryngoscopy and intubation, and （4） cough- ing and movement of the endotracheal tube on the larynx during the endotracheal tube in place. It was sug- gested that correct technique and gentle manipulation during intubation could reduce the incidence of the complication. Timely postoperative follow-up and early reduction of arytenoid cartilage should be empha-sized in the diagnosis and treatment of the complication.Keywords：arytenoid cartilage;dislocation;intubation4|11|0Updated：2024-04-10
- Abstract：We determined and analysed the ApoE polymorphism of 30 sporadic Alzheimer’s disease （AD） pa- tients, 27 patients with multi-infarct dementia （MID） and 46 aged healthy subjects as control. The results showed that the frequency of ApoE E4/3 genetype in AD group was significantly higher than that in con- trol （P<0. 05）. Among these three groups, ApoE 4 allele frequency in AD group was significantly higher than that in control （P<0. 01 ） and MID group （P<0. 05）. Among the three ApoE alleles, the risk ratio of ApoE E4 allele in AD group was 4. 114（p<0. 01 ）. There was statistically significant （P<0. 05） as the increasing of ApoE 4 gene dose in AD. It suggests that ApoE is related to AD of Chineses and it might be a genetics index of early diagnosis for AD.Keywords：apolipoprotein E;polymorphism;Alzheimer’s disease4|7|0Updated：2024-04-10
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