Objective To investigate the genetic association of the PIP3-E locus in the long arm of chromosome 6 with schizophrenia among 89 Chinese Han family trios, consisting of fathers, mothers, and affected offsprings with schizophrenia. Methods Genomic DNA was abstracted from the whole blood samples. The PCR-based restriction fragment length polymorphism (RFLP) technique was used to detect the association between single nucleotide polymorphism（SNPs）of rs2236257 and schizophrenia. Results ①The distribution of genotype frequencies of rs2236257 was not deviated from the Hardy-Weinberg equilibrium; ②The haplotype relative risk test (HRR) did not show a genetic association between rs2236257 and schizophrenia (P>0.05); ③The transmission disquilibrium test (TDT) did not show significantly biased transmission from parents to affected offsprings（P>0.05）; ④There were correlations between rs2236257 allele frequencies and some clinical symptoms of schizophrenia,such as incoherence of thinking,poverty of thought and premorbid personality (P<0.05). Conclusion The polymorphism of rs2236257 in the PIP3-E locus may not be associated with schizophrenia although the association of the other SNPs at the PIP3-E locus with the illness can not be ruled out.

Objective To clone the sequence of the cDNA of mouse interferon-gamma coding area and construct radiation-inducible expression plasmid pIRESEgr-IFNγ containing Egr-1 promoter. Methods With the technique of RT-PCR, mouse splenocyte mRNA was used as template to obtain full length mouse IFNγ. The pGEMT-IFNγ was sequenced automatically and radiation-inducible expression plasmid pIRESEgr-IFNγ containing Egr-1 promoter was constructed with gene recombinant technique. Results The sequencing proved the cloned mouse IFNγ cDNA to be completely identical with that reported in the literature and the radiation-inducible expression plasmid pIRESEgr-IFNγ containing Egr-1 promoter was constructed successfully. Conclusion Mouse IFNγ cDNA was cloned and the radiation-inducible expression plasmid pIRESEgr-IFNγ containing Egr-1 promoter was constructed successfully.

Objective To study the proliferation of hematopoietic cell in patients with myelodysplastic syndrome (MDS). Methods Liquid single layer culture and semisolid culture in vitro were used and flow cytometry was used to detect the cell cycle, apoptosis and the expressions of PCNA, P53,and BCL-2. Results ①The Clony Forming Unit-Granulocyte-Macroplage(CFU-GM) and the Erythroid-Clony Forming Unit(CFU-E) of myelodysplasyic syndromes (MDS/RA) decreased significantly (P<0.001); while CFU-GM,CFU-E of MDS/RAEB could not be detectable; ②The percentage of G1 phase increased and the percentage of S, G₂/M phase decreased significantly (P<0.01), compared with control group; ③The increase of apoptotic bodies of MDS/RA was much more markedly (P<0.01), while the changes of those in MDS/RAEB could not be found significantly; the expression of PCNA of MDS increased in both MDS/RA and MDS/RAEB (P<0.01), especially in MDS/RAEB group;④the protein expression of P53 in MDS increased, while BCL-2 decreased significantly in MDS, compared with normal control (P<0.01). Conclusion There are severe abnormalities in hematopoietic cell of patients with MDS, higher proliferation and excessive apoptosis occur in bone marrow cells and the changes of p53,bcl-2 gene suggest their possible participation in the regulation of apoptosis.

Objective To detect the association between metallothionein (MT) gene polymorphism and type 2 diabetes mellitus (T2DM) in Chinese Han people. Methods The techniques of PCR and restriction fragment length polymorphism (RFLP) were used to examine MT4 gene (rs666636) G→A mutation in 41 patients with T2DM and 53 unrelated healthy individuals, and then the mutation frequency was statistically computed. Results The alleic frequency (A) of MT4 gene mutation was significantly different between the diabetic and control group (P<0.05,OR=2.335).The distribution rates of AA, GA and GG had also remarkable differences (P<0.05).The relative risk of suffering from T2DM of GA genotype was 3.462 times (OR=3.462，95% OR CI=1.389~8.629) GG genotype. Conclusion There′s an association between the alleic gene of MT4 gene mutation and T2DM, and the mutation genotype has increased the risk of suffering from T2DM.

Objective To isolate and culture the rabbit bone marrow mesenchymal stem cells (MSCs) and then to induce MSCs into chondrocytes in vitro. Methods The MSCs was isolated and expanded from cultured and nucleated cell fraction of autologous bone marrow. Dexamethasone(10_-7 mol•L^-1), ascorbic acid(50 mg•L^-1) and basic fibroblast growth factor (bFGF 10 μg•L^-1) were added into MSCs medium for the first week. Then the bFGF was replaced by transforming growth factor-β₁ (TGF-β₁ 10 μg•L^-1) for next three weeks. Results Some spindle MSCs turned into round or elliptic shape and secreted metachromatic matrix with Toluidine blue. The alkaline phosphates(ALP) activity of MSCs increased to twenty fold . The RT-PCR indicated that the MSCs mainly expressed type-Ⅰ collagen mRNA before induction. After induction the partial differentiated cells begun to express type-Ⅱ collagen mRNA.Conclusion In this condition the rabbit MSCs may be induced into chondrocytes in vitro.

Objective To investigate the inhibitory effects of scorpion venom (SV) and its separated components (SV-Ⅰ、SV-Ⅱ and SV-Ⅲ) on the growth of B16 melanoma cells. Methods MTT method was used to detect the survival of tumor cells treated with SV and its components in vitro and colony forming assay was applied to measure the proliferation of tumor cells. Moreover, flow cytometry was further adopted to observe the changes of cell cycle of tumor cells. Results SV-Ⅰ, SV-Ⅱ and SV-Ⅲ could inhibit the growth of B16 melanoma cells significantly at the concentration of 200, 400 and 800 mg•L^-1 (P<0.05~P<0.01) and SV could inhibit the growth of B16 melanoma cells at 400 and 800 mg•L^-1 (P<0.01~P<0.05). Furthermore, SV and its components could inhibit colony forming of B16 melanoma cells (P<0.01) at 400 and 800 mg•L^-1. Cells in G₀/G₁ stage were decreased and cells in G₂+M stage were increased obviously (P<0.05~P<0.01) after　SV-Ⅰ, SV-Ⅱ and SV-Ⅲ were given at 400 and 800 mg•L^-1, respectively. Conclusion SV and its components can inhibit the proliferation and survival of B16 melanoma cells. Moreover, the inhibitory effects of components are better than those of SV on B16 melanoma cells. Changes of cell cycle will be an important mechanism of inhibitory effects of SV.

Objective To study the effects of serum on p21^WAF1 expression during U937 cell differentiation. Methods U937 cell differentiation induced by PMA (Phorbol Ester) was used and divided into four experiment groups according to whether or not PMA and serum exsited in culture system: serum group (FCS), serum plus PMA group(FCS+PMA), serum free group(NOFCS), serum free plus PMA group (NOFCS+PMA). FCS group was used as control group. Cell adherence was identified as differentiation characteristics,p21^WAF1 protein expression was determined by Western Blot. Results After PMA was used for 24 hours, cell stick growth ,U937 cell adherence and p21^WAF1 protein expression in FCS+PMA group increased significantly compared with control group. Cell suspend growth and U937 cell adherence in NOFCS and NOFCS+PMA groups were no significant difference compared with control group. p21^WAF1 protein expression in NOFCS+PMA group increased significantly, but that in NOFCS had no significant difference compared with control group. Conclusion Serum can′t affect p21^WAF1 expression during U937 cell differentiation induced by PMA,but it can affect cell differentiaiton maturation.

Objective Whether amniotic epithelial cells could promote the survival and differentiation of neural stem cells was explored in vitro. Methods Neural stem cells were dissociated and cloned from the cortical tissue of E12-14 d rats. Amniotic epithelial cells were also dissociated and purified simultaneously from the amnion. The neural stem cells and amniotic epithelial cells were co-cultured. The expression of marker of neural stem cells and differentiated cells (neuron and glial cell) in the neural stem cells and amniotic epithelial cells were detected by immunohistochemistry method. The morphological changes of neural stem cells were examined with light microscope. Results Compared with control group,the total differentiated cells and percentage of MAP-2 positive cells was significantly increased in co-cultured groups. The primary neurite of neuron-like cells in co-cultured groups were obviously larger than that in control group. Conclusion Amniotic epithelial cells can provide a supportive environment for the survival and neuronal differentiation of neural stem cells and suggest that their addition may be useful as a sustained source of trophic support to improve outcome of neural stem cells transplantation.

Objective To study the relationship of donor-derived NK cells as pretreatment condition and immune tolerance after MHC haplotype-mismatched BMT in mice. Methods The murine model of MHC haplotype-mismatched BMT was established by using (BALB/c^H-2d×C57BL／6^H-2b) CB6F₁^H-2d/b mouse as the recipient, and C57BL／6^H-2b mouse as the donor. Seventy recipient mice were divided into seven groups after irradiation (TBI 60Co 9.0　Gy). Control groups: simple-irradiation group, MHC haplotype-mismatched GVHD-control group, MHC haplotype-mismatched BMT group, syngeneic BMT group. Experimental groups: the mice were distributed into three groups of different doses of NK cells which were 1×10⁶/one，5×10⁵/one，2×10⁵/one. CB6F1^H-2d/b mice were conditioned with 9.0 Gy, and the donor-derived NK cells were injected into CB6F₁^H-2d/b mice, after a interval of four hours followed by infusion of C57BL／6^H-2b mice bone marrow cells. The effect was assessed by hemotopoietic reconstruction, survival time, body weight, histopathology in the recipients.　Results ①Life span: in the case of control group, survival time was (5.15±0.66) days for the simple-irradiation group, survival time was (15.80±1.93)days for MHC haplotype-mismatched GVHD-control group, beyond 30 days for other groups. ②Pathological results: GVHD pathological manifestations were found in MHC haplotype-mismatched GVHD-control group. The effects in other cases assessed by histopathology indicated no GVHD. Conclusion The results have shown that donor-derived NK cells as pretreatment condition can induce immune tolerance after MHC haplotype-mismatched BMT in mice.

Objective To determine the effect of morphine on RNA synthesis of C6 glioma cells. Methods Gene expression level of TFⅡF was detected by RT-PCR. Results Exposure of morphine to C6 glioma cells caused a decrease of TFⅡF gene expression as compared with corresponding control cells, and the effect could be reversed by naloxone. Conclusion The down-regulation of morphine on gene expression of TFⅡF may be mediated by μreceptor.

Objective To observe the effects of high concentration glucose on bFGF expression and calcium channels of rabbit retinal Müller cells cultured in vitro. Methods Transmission electron microscopy and immunocytochemical staining method were used to identify Müller cells. The changes of bFGF expressed by Müller cells were studied with immunocytochemical method in a qualitative way. Patch-clamp technique was used to observe the changes of calcium channels caused by high concentration glucose. Results bFGF expression was enhanced obviously by high concentration glucose. Obvious changes of calcium channels were not found. Conclusion bFGF expression secreted by cultured Müller cells in vitro can be enhanced at high concentration glucose. Müller cells play a potential role in neovascularization of diabetic retinopathy. Obvious changes of calcium channels are not found during the process.

Objective To study the effect of diosgenin on the cell diversion and colony forming of human poorly differentiated gastric adenocarcinoma cell line MGC-803,and investigate the mechanism of cell death induced by diosgenin. Methods MTT assay,colony forming experiment and ［³H］-TdR incorporation into DNA were used. Results After incubated with diosgenin for 24, 48, and 72 hours, the IC₅₀ were 56.290, 27.837, and 17.723 mg•L^-1,respectively, detected by MTT. The concentration which inhibited half colony forming was 5.486 mg•L^-1. It directly inhibited the synthesis of DNA of MGC-803 in lower concentrations (3.750 and 7.500 mg•L^-1). Conclusion Diosgenin has an obvious inhibitory effect on the cell diversion and colony forming of MGC-803.

Objective To investigate the analgesic effect of Gutongling ointment (GTLO) and its effect on viscidity of blood. Methods Analgesic effect of GTLO and its effect on viscidity of blood were carried through ache models induced by wring body and hot board methods in mice, by irradiation method of radiant heat in rats and test of viscidity of blood in rats with acute stasis of blood. Results GTLO had significant inhibitory action on ache models induced by wring body and hot board methods in mice and by irradiation method of radiant heat in rats, moreover it could reduce significantly viscidity of plasma in rats with acute stasis of blood. Conclusion GTLO has significant inhibitory effect on many experimental ache models and can improve significantly viscidity of plasma in rats with acute stasis of blood.

Objective To discuss the effects rhidosin in expression of Bcl-2 and Bax genes related with apoptosis in olfactory bulbs of senile rats and its significance. Methods Twenty healthy rats were randomly diveded into rhidosin injected group (n=10) and physiological saline control group. After decaptation the olfactory bulbs were immediately injected with neutral formalin, followed with paraffin-embedding, serial sectioning, immunohistochemical staining and light microscopic observation. Results Bcl-2 and Bax mainly expressed on mitral cells of olfactory bulbs in senile rats. The positive expression rate of Bcl-2 gene in the rhidosin group was significantly higher than that in the physiological saline control group (P<0.01); the positive expression rate of Bax gene in the rhidosin group was slightly lower than that in the physiological saline control group (P>0.05). Conclusion Rhidosin can increase the expression of Bcl-2 protein in olfactory bulbs of senile rats, inhibit the apoptosis of mitral cells and has the effect of antiapoptosis in olfactory bulbs of senile rats.

Objective To construct recombinant Murine IL-18 (mIL-18) plasmid which could be expressed in eukaryotic systems and be used as immunoadjuvant. Methods mIL-18 was amplified from BALB/c and Swiss mouse liver mRNA by RT-PCR, and ligated orientatively into the eukaryotic expressing plasmid pcDNA3. The recombinant plasmids, pcIL-18,were evaluated with endorebonuclease digestion and sequencing. Results Two recombinant plasmids were constructed. The sequence of mIL-18 from BALB/c mouse was consistent with that of Genbank, while the one from Swiss mouse had two point mutations. Conclusion Two recombinant plasmids pcIL-18 are constructed successfully. They will play a key role in reinforcing DNA vaccine. And gene polymophism in mIL-18 has been found and received by Genbank (AY362457).

Objective To investigate the changes of purine nucleotides catabolism on morphine-dependent and withdrawal condition and its mechanism. Methods Rats were administered with morphine by intraperitoneal injection with increasing doses to establish addiction models.The level of uric acid in plasma and the activities of xanthine oxidase (XO) in the plasma and the small intestine were determined. RT-PCR was used to examine the gene transcripts of key enzyme of purine nucleotides catabolism ,XO mRNA. β-actin was used as control gene in RT-PCR study.Results The level of uric acid in plasma of morphine-administered group was significantly higher than that of control group (P<0.05).The activity of XO in plasma of morphine-administered group was higher than that of control group (P<0.05),as well as natural withdrawal group and naloxone control group (P<0.01). The activities of XO in the small intestine of morphine-administered group and naloxone withdrawal group were extremely higher than that of control group (P<0.05). The levels of transcript of XO mRNA of morphine-administered group, natural withdrawal group and naloxone withdrawal group were higher than that of control group. Conclusion Morphine can promote the purine nucleotides catabolism.Morphine may increase the activity of the key enzyme of purine nucleotides catabolism in the small intestine, which is related to the increasing of gene expression. Morphine can induce XO synthesis in mRNA level, naloxone can′t obstruct it in the short time.

Objective To explore the antagonism of ginsenoside to the toxicity of embryo development induced by acetic lead in rat. Methods The embryo teratogenic model in rat was established by whole embryo culture in vitro, antagonistic experiment was made with ginsenoside. Results Acetic lead had stronger developmental toxicity on rat embryo and the relationship of dose-effect was demonstrated, the developmental toxicity on rat embryo could be antagonized by ginsenoside. Ginsenoside could ensure normal growth of rat embryo. Conclusion Ginsenoside has better antagonism on embryo teratism in rat induced by acetic lead and can proportionately promote the differentiation and development of rat embryo.

Objective To induce the prolongation of survival period of heart allografts in recipient rats by oral administration of donor spleen cells,and to study the mechanism of oral tolerization. Methods The inbred SD rats were chosen as heart donors and inbred Wistar rats as recipients,heterotopic heart transplantations were performed. Sixteen recipients were divided into group A (control group,only performing heterotopic heart transplantation) and group B(feeding group,feeding on 5×10^7 donor splenocytes by means of gastric intubation each day). After 7 days, the mixed lymphocyte culture (MLC) in vitro and delayed-type hypersensitivity response (DTH) in vivo were examined, and the allogenic heart transplantation was performed, then the survival time of heart allograft was observed.　 Results MLC in vitro and DTH in vivo existed antigen specific reduction after rats were fed on alloantigen orally. The survival period of heart allografts in group A and group B were (7.0±0.8)d and (12.1±1.9)d ,respectively. Conclusion Oral administration of alloantigen can down-regulate the immune response to histocompatibility antigens, prolong the survival times of heart allografts.

Objective To study the effect of hemimastication on the growth of mandible in rat. Methods Rat models with hemimastication were built up, and sacrificed 8 weeks later. Mandibula were resected for soft X ray radiographing and pointing, then the distances between the points were meatured with graph analysis system. Results As for the lengthes of mandibula and mandibular bodies in experimental group increased compared with control group, and the non-mastication side increased compared with mastication side in experimental group; the condyloid process heights of mastication and non-mastication side in experimental group were lower than those in control group; the coracoid height of mastication side in experimental group was obviously higher than that in control group. While between mastication side and non-mastication side, non-mastication side and control group, no differences could be found. The results of the height of mandibular angle measuring showed that there were some differences between the mastication and non-mastication side in experimental group, the height of mandibular angle of mastication side was lower than that of control group, and the height of mandibular angle of non-mastication side was higher than that of control group. Conclusion Hemimastication may contribute to the asymmetry growth of mandibula and mandibular angle, deficient growth of the condyloid process, which suggesting that hemimastication can influence the growth of mandibula.

Objective To investigate the suitable method to cultivate the adult human endothelium from the great saphenous vein. Methods Endothelial cells (EC) from small discarded segments of human great saphenous vein of patients undergoing coronary artery bypass surgery were digested with 2% collagenase Ⅱ in medium 199,seeded into culture flask with high density about (1.5×10⁴/cm²),grew in medium containing the 10%~20% human serum collected from the same patient. After reaching confluence, the EC were detached with a 0.25% trypsin-ethylenediamine tetraacetic acid (EDTA) solution (1 mmol•L^-1),and seeded into culture flask with low density about (0.8×10⁴/cm²),　subcultivated as described for primary cultures　(split ratio 1∶3). Results Primary endothelial cells were confluenced about (12.0±2.3)d and 7.0×10⁴/cm² EC or more could be achieved,the growth of passage cells were more exuberant and confluenced about (7.0 ±1.1) d. The EC retained spindle-shaped or cobblestone appearance,Weibel-Palade (WP) could be found by the transmission electron microscopy, and CD31 antigens were positive in the EC by the immunofluorescent histochemistry. Conclusion The results suggest that it is a useful method for study on cultured endothelial cells and tissue engineering.

Objective To establish solid intracerebral human glioma model in Wistar rat with xenograft methods. Methods The SHG-44 cells were injected into brain right caudate nucleus of previous immuno-inhibitory Wistar rats with stereotactic technique. The MRI scans were performed at 1 week and 2 weeks later after implantation. After 2 weeks the rats were killed and pathological examination and immunohistologic stain for human GFAP were used. Results The MRI scan after 1 week of implantation showed the glioma was growing,pathological histochemical examination demonstrated the tumor was glioma. Human GFAP stain was positive. The growth rate of glioma model was about 60%. Conclusion Solid intracrebral human glioma model in previous immuno-inhibitory Wistar rat is successfully established.

Objective To study the method to isolate and identify Actinobacillus actinomycetemcomitans (A.a) and insure its resistance to antibiotics. Methods A.a was isolated from specimens of human periodontal disease, and identification of A.a was carried out by anaerobic cultivation, observation of growth properties on TSBV and systematically biochemical reaction; and the spectrum of A.a′s resistance to antibiotics was identified. Results Twenty-four strains of A.a were acquired and identified; They were sensitive to Re，MTZ，Sm，Em，and Cef. Conclusion Re，MTZ，Sm，Em, and Cef can play a important role in treatment of peridontitis.

Objective To establish the animal model of periodontitis. Methods Fifty-one two-month-old female white rats were selected and divided into four groups randomly: control group, hormone group, ligation group, ovariectomized and ligation group. The control group was treated with intramuscular injection of 0.4 mL 0.9% NaCl each day. The hormone group was treated with intramuscular injection of 1 mg dexamthasone and 0.4 mL 0.9% NaCl each day. For the ligation group, the white rat′s maxillary first molar was ligated by orthodontics wire around the cervical. In the ovariectomized and ligation group, the same tooth was ligated by orthodontics wire after being bilateral overiectomized. The formation of periodontitis in each group was observed with histopathological method and determination of alveolar bone density. Results The results demonstrated that after bilateral ovariectomized, the rat′s alveolar bone density decreased significantly. The hormone group showed the clinical syndrome-so called “kidney deficiency” in traditional Chinese medicine. In ligation group the local bacterial plaque accumulated around the periodontal tissues and resulted in the mechanical and inflammatory stimulate changes. In the ovariectomized and ligation group, the pathological changes resembling the human periodontitis were observed in the rat models, such as the decreased alveolar bone density, the absorbed and destructured alveolar bone, the loose tooth and the formatted pocket. Conclusion Periodontitis models were established by means of the ovariectomized and ligation.

Objective To investigate the method to establish fatty liver model of rats. Methods The rats were fed on a diet deficient in choline and amino acid (methionine in chief) and fatty liver models were established successfully. The changes of rat portal vein pressure, ratio of liver weigh/body weigh, and pathological structure of liver were observed. Results Fourteen days later, much more fatty droplets were accumulated in the livers of rats, more than two thirds of liver cells had fatty droplets accumulation after twenty-one and twenty-eight days. The form of fatty liver cells were irregular, liver sinus became narrow and irregular; portal vein pressure of the fatty liver was higher than that of normal one, weights of the liver and body were both lighter than that of normal one. Conclusion This method is effective and economical to establish fatty liver model of rat in short time.

Objective To establish a new culture system for retinal ganglion cells (RGCs). Methods Four male Wistar rats aged 10 weeks, retina was isolated from the choroid, the pigment epithelium and adherent vitreous. The retinal explants from mid-peripheral retinas were dissected into 16 pieces (approximately 0.5 mm×0.5 mm square). Four pieces of retinal tissues were embedded in the type I collagen gel with the nerve fiber layer downward on plastic dish. Each dish was pre-coated with 10 mg•L^-1 poly-L-lysine. The explants were cultured in serum-free MEM and maintained at 37℃ in an incubator under a 5% CO₂/95% air atmosphere. The number of outgrowing neural protuberances was counted under phase contrast optics using inverted microscope. Immunohistochemical markers, monoclonal antibodies against neurofilament and Thy 1.1 were used to identify the nature of the outgrowing processes. Results Using the three-dimensional culture, the retinal explants were stably embedded just below the surface of the gel, and they could be cultured for 2 weeks or more. The outgrowing neural protuberances could elongate more freely into gel in the three-dimensional culture than in the two-dimensional one. Conclusion RGCs can be cultured sucecessfully in the three-dimensional culture.

Objective To discuss reversal of multidrug resistance (MDR) of scorpion venom of buthus martensii kirsch (BMK) in MCF-7/ADM cell line. Methods MTT colorimetric method was used to detect the IC₅₀ of MCF-7/ADM cell line when BMK was used in combination with adriamycin (ADM). Fluorescence spectrophotometer method was applied to detect the effect of BMK scorpion venom on intracellular ADM concentration of MCF-7/ADM cell line. Results BMK in different groups (100~350　mg•L^-1) enhanced the sensitivities of MCF-7/ADM cells to ADM (Ｐ<0.05;Ｐ<0.01). BMK in different groups (100~200 mg•L^-1) could markedly increase the intracellular ADM concentration of MCF-7/ADM cell line. Conclusion The results suggest that BMK can partly reverse the MDR of MCF-7/ADM cell line to ADM.

Objective To study the applied anatomy of the pudendal nerve and provide theoretical basis for clinical work. Methods After dissected in buttocks, the course, diameters and body surface projections of pudendal nerve were observed in 12 lateral adult cadavers. Results The pudendal nerve′s location was about 12 mm medialis to the middle and lower 1/3 point of the line between posterior superior iliac spine and ischial tuberosity. The maximal free length of the pudendal nerve was (38.74±1.39)mm from the start point to the superior border of sacrotuberous ligament. Conclusion The defecation,miction and sexual function may recover by suturing the pudendal nerve with the peripheral nerve above the spinal injury plane in paraplegia patients.

Objective To observe the role of platelet-derived growth factor-B (PDGF-B) in the regeneration of peripheral nerve by studying its expression on injured rat sciatic nerves. Methods The rat sciatic nerves cut and crush-injured models were established, the expressions of PDGF-B on both injured nerves were detected by immunohistochemistry technique. Results The expressions of PDGF-B were increased in proximal nerve segments and regenerating axons in both models. Soon after both types of injury, considerable amounts of PDGF-B accumulated in Schwann cells in distal segments of both models. With restoration of the axon-Schwann cells relationship in the crush model, levels of PDGF-B tended to decrease, eventually returned to normal. In the cut model the relationship can not be restored, the PDGF-B was decreased to a very low level. Conclusion The increased expression of PDGF-B after sciatic nerve injury may contribute to peripheral nerve regeneration.

Objective To investigate the expression of the protein p27^kip in the tissue of oral squamous cell carcinoma (OSCC) and study the relationship between it and the generation and development of OSCC. Methods The expression of the protein p27^kip was detected by immunohistochemistry S-P in 37 cases of OSCC, 30 cases of the tissue beside OSCC and 10 cases of normal oral mucous membrane. Results The positive expression rates of protein p27^kipin 37 cases of OSCC, 30 cases of the tissue beside the OSCC and 10 cases of normal oral mucous membrane were 67.6%, 93.5%, and 100%, respectively. There was a remarkable difference between them (P<0.01). The expression of the protein p27^kip was decreased with the increase of the pathological classification and clinical stage of OSCC. Conclusion The expression of the protein p27^kip is related with the generafion, development and the degree of OSCC. It is considered a valuable marker in evaluating the prognosis of the patients with OSCC.

Objective To study thrombopoietin (TPO) in the expansion of cord blood hematopoietic stem/projenitor cells in vitro. Methods In the colony culture system and the liquid expansion in vitro, different cytokines combination groups with or without TPO were used to observe the result. Results The proliferation folds of colony BFU-E and colony CFU-GM, the total cells and CD34⁺ cells in the groups with TPO were in obviously higher than those in the groups without TPO (P<0.05). Conclusion TPO can improve the expansion effect of the early hematopoietic stem/projenitor cells obviously with SCF,IL-3 and other traditional cytokines.

Objective To study the killing effects of induced LAK-cells on B₇ gene transfected cells of liver carcinoma. Methods A new vector pRCCMV-B_7-1 containing B_7-1 gene was constructed to establish the new cell line B₇⁺SMMC7721 through transfection B₇ gene into the hepatocarcinoma cell SMMC7721 by liposome-mediated transduction. RT-PCR and flow cytometry were used to identify the expression of the transfected gene. MTT assay was used to evaluate the killing effects of LAK cells activated by IL-2 on the transfected line and the wild type of the cells.　Results B₇ gene was steadily expressed in B₇⁺SMMC7721 and LAK cells had effectively killing effects on the B₇⁺SMMC7721 cells. Conclusion B₇ gene can be transfected into hepatocarcinoma cells and can be expressed steadily in vitro, which can increase the efficiency of LAK cells activated by IL-2 on them.

Objective To find the relationships between endothelin （ET） and insulin resistance　(IR) and insulin receptor　(INSR) in patients with essential hypertension. Methods Forty patients including 20 cases of essential hypertension disease (EHD) and 20 health persons were divided into experimental group and control group. Blood glucose,serum insulin,ET and the number of erythrocyte INSR in all patients during fasting condition were detected by radioimmunoassay and radiometric analysis. Results Both insulin sensitivity index（ISI） and the number of INSR in EHD group were much less than that of control group, on the contrary, ET level of EHD group was significantly higher than that of control group (P<0.05). Statistical analysis demonstrated a negative correlation between ET and ISI and INSR number existed in EHD group. Conclusion IR is a common phenomenon in patient with EHD and possiblely due to decrease of INSR number. The ET levels are higher in patients with EHD than that in health people and correlate with INSR, and the change of INSR number is the possible mediator for their relationship.

Objective To investigate the association of beta 2 adrenergic receptor (β₂AR) gene with asthma in children. Methods The 5′ 234 bp fragment of β₂AR gene was analyzed by PCR method. Results The fragment deletion of β₂AR existed in 22 cases of 65 asthmatic patients and in 6 cases of 20 healthy clildren in control group, the deletion rates were 33.8% and 30%，respectively . There was no significant difference between the two groups. The deletion rate of β₂AR gene fragment in severe asthma group was higher than that in mild-moderate asthma group. Conclusion The deletion of this fragment of β₂AR gene isn′t related to onset of asthma, but it is related to onset of severe asthma.

Objective To study the role of vascular cell adhesion molecules (VCAM-1) and tumor necrosis factor （TNF-α）on the pathogenesis of pregnancy-induced hypertension (PIH). Methods ELISA and radioimmunoassay were used to measure the levels of VCAM-1 and TNF-α in maternal serum and cord blood of 20 cases of normal pregnant women and 60 cases of PIH women. Results The serum levels of VCAM-1 and TNF-α in the moderate and severe PIH group were significantly higher than that in control group (P<0.05). There was no difference of the levels of VCAM-1 and TNF-α in cord blood between PIH and control group. There was significant relativity between VCAM-1 and TNF-α in serum of PIH patients (P＜0.05). Conclusion VCAM-1 and TNF-α may be important factors in the pathogenesis of PIH.

Objective To study the relationships between plasma adrenomedullin (ADM) and normal pregnancy and pathogenesis of pregnancy-induced hypertension(PIH) . Methods ADM concentrations in the plasma from 10 normal non-pregnant women,36 normal pregnant women (12 first,12 second,12 third trimester, respectively) and 30 cases of PIH (10 mild,10 moderate,10 severe, respectively) were determined by radioimmunoassay, and data were analyzed statistically. Results ADM concentrations in the first, second and third trimester of normal pregnancy increased significantly than that of normal non-pregnant women (P<0.05). ADM concentration in the plasma of patierts with PIH was higher than that of third trimester pregnancy (P<0.01). There were significant differences between mild, moderate, and severe PIH groups (P<0.05). In the PIH groups, significant positive correlation was found between plasma ADM concentration and mean arterial pressure(r=0.822,P<0.05). Incidence of low birth weight infants was related to serious degree of PIH. Conclusion ADM may involve in maintaining normal human pregnancy. ADM may increase compensatorily in the pathogenesis of PIH.

Objective To establish the purified cell cultures of stromal cell of endometrium in vitro. Methods Collagenase digestion,trypsase digestion,light microscope,and immunocytochemical staining were used. Endometrium and endometriotic tissues were isolated, purified,and cultured. Results The purity of stromal cells was 95% identified by cytokeratin and vimentin staining. Conclusion The purified stromal cell cultured from endometrial and endometriotic tissues are obtained by centrifugation. The cellular model will be useful to investigate the pathogenesis and therapy of endometriosis.

Objective To study the relationship between nitric oxide (NO) and apoptosis in the focal cerebral ischemia. Methods Using light microscope and electric microscope,the pathological changes of cerebral in the focal cerebral ischemia were observed. Results When the focal cerebral ischemia in rats was influenced by high dose N-nitro-L-arginine ( NNLA ), the operation side brain appeared edema around nucleus, dissolutuin and distortion of neuronal cell nucleus in a part of hippocampus CA4 region and cortex region. These changes under hight microscope were more obvious than that in operation control group. Under electric microscope, the changes of the organelles of neuronal cell nucleus and chromatin dense stain of gliocyte nucleus in ischemic penumbra were found. Conclusion Excessive NO has two forms to induce neuron injury. First,excessive NO is a reactive oxygen species (ROS). It may attack straightly to nerves and vascular endothelial cell and is a direct toxicity molecule. Second, the lack of NO can not keep the cerebrovasurlar basic tension. It may induce the decrease of cerebral blood flow and anoxia and neuron injury.