Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690-Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 inE. coli was very low, but that of CH63 was very high. The results suggests that Asp1961 -Glu1978 in FN is a key sequence influencing the expression of triple-domain polypeptide inE. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography. Both of the cell-binding domains in the recombinant polypeptide were functional. The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function.

Oxidatively modified low density lipoprtein (LDL) plays an important role in atheroslerosis (AS) development. To investigate the role of neferine (Nef) in anti-LDL oxidation and foam cell formation, the lipoprotein was derived and subjected to three different treatments: N-LDL (normal LDL), Cu²⁺+LDL and Cu²⁺ + Nef+LDL. The LDLs were put at 25 C for 24 h and the thiobarbituric acid reactive substance (TBARS) values were determined. They were 0. 57 ±0. 02, 6. 01 ±0. 22 and 2. 26±0.13 nmol/mg protein, respectively. The difference was very significant (P<0. 01) for each two groups byt test. Mouse peritoneal macrophage (MΦ) were exposed to 50 μg protein/ml of Cu²⁺+LDL and Cu²⁺+Nef+LDL at 37 °C for 60 h. The tryglyceride (TG) and total cholesterol (TC content in MΦ were assayed. The results showed that Cu²⁺+ LDL was more efficient than Cu²⁺+Nef+ LDL in stimulating lipid accumulation in MΦ(P <0. 001). The study demonstrated that Nef could inhibit Cu²⁺-mediated LDL oxidation and thereby inhibiting macrophage-derived foam cell formation.

The whole-cell patch-clamp technique was employed to obtain information about the voltage-dependence and kinetics of interaction of 7-chlor-benzyltetrahydropalmatine (7-C1-BTHP) with cardiac sodium channels. 7-C1-BTHP (30 mol/L) significantly decreased the peak sodium current (from 7. 8±1. 8 nA to 5. 3±1. 4 nA,P<0. 01,n = 5), without producing a shift of the current-voltage curve. It shifted the inactivation curves of sodium current to hyperpolarized potentials, and the V_0.5 was shifted from - (82. 5±2. 5) mV to - (95±2. 4) mV (P <0. 05,n = 4). 7-C1-BTHP produced a significant use-dependent effect that was proportional to the duration of the voltage step. In addition, 7-C1-BTHP slowed the recovery of sodium channel from inactivation, which could explain its use-dependent effects on sodium current. The characteristics of 7-C1-BTHP blockage suggest that this agent binds preferentially to inactivated sodium channels.

Abnormality of ras gene family was studied in a total of 206 cases of gastric cancer and precancerous lesions by PCR-RFLP, PCR-SSCP and DNA sequencing. The results showed that mutation rate of H-ras 12 codon in metaplasia, atypical hyperplasia, early-stage cancer and advanced cancer was 16.7%, 31. 2 %, 50. 0 %, and 32. 2 %, respectively. In the groups of superficial gastritis and normal controls, no mutation were detected in codon 12 of ras. Mutations of H-ras 61 codon and N-ras 12 codon in various groups were the same as those in normal control. K-ras 12 codon mutation was detected in only 2 cases of gastric cancer by using PCR-SSCP, but it was not detected by DNA sequencing, which may be polymorphism. All H-ras 12 codon mutations were G→ T mutation. There were significant difference between the groups of metaplasia, dysplasia, gastric carcinoma and normal control group (P < 0.05,P < 0.01,P < 0.01, respectively). It was concluded that H-ras 12 codon mutation was an early event and may play an important role in gastric carcinogenesis. Although K-ras, N-ras mutation rates are high in colon cancer and leukemia, it seems to bear no relationship with gastric cancer.

HSP₆₀ HSP₇₀ in plasma of 11 cases of Kawasaki diseases (KD) and 23 healthy children were determined. The two groups were controlled for age. Determination of HSP₆₀, HSP₇₀ was conducted in lymphocytes of 14 cases of KD and 26 healthy children. The results were compared with those of 12 patients with febrile diseases and 10 patients with tuberculosis. Our results showed that except a significant difference in plasma HSP₇₀ found between acute phase and convalescent phase of KD (P<0.01), no significant difference was found in HSP₆₀, HSP₇₀ among all groups (P>0.05). The differences in HSP₆₀, HSP₇₀ in lymphocytes were relatively obvious among all groups. The levels of HSP₆₀, HSP₇₀ in acute phase of KD were significantly higher than those in convalescent phase or in healthy controls (P<0.01). The levels of HSP₆₀ in KD were significantly higher than those of patients with febrile diseases. HSP₆₀ of KD children was significantly lower than those of children with tuberculosis (P< 0.01). The findings showed that HSP₆₀, HSP₇₀ might contribute to the pathogenesis of KD. Determination of HSP₆₀ HSP₇₀ in lymphocytes is of help in the diagnosis of KD.

In order to determine the replication sites of hepatitis C virus, thein situ hybridization and immunohistochemical technique using digoxin-labeled 531bp plus-strand and minus-strand HCVRNA probes were employed to detect HCV-RNA in the liver tissues, bone marrow mononuclear cells and peripheral blood mononuclear cells (PBMCs) from the patients with chronic hepatitis C, and in HCV transfected COS cells. The results showed that both plus-strand and minusstrand HCVRNA were detected in 80 % of liver tissues (4/5). Plus-strand HCVRNA could be detected in 90 % of PBMCs and bone marrow mononuclear cells (18/20), minus-strand HCVRNA in 25 % of PBMCs. In HCV transfected COS cells, plus-strand HCVRNA distributed evenly in 20 % cellular nuclei and cytoplasms. No minus-strand HCVRNA was detected in the bone marrow mononuclear cells and HCV transfected COS cells. The positive signal appeared in more cells when the liver tissues, PBMCs and marrow mononuclear cells were hybridized by plus-strand probes than when hybridized by minus-strand probes. Our results suggested that the hepatocytic cytoplasms and PBMC cytoplasms were the replication sites of HCV, but the marrow mononuclear cells were not the replication sites of HCV although they were infected by HCV. HCV infection might be accounted for the pathogenesis of chronic hepatitis and relapse of hepatitis C after liver transplantation.

Immunoperoxidase histochemical assay with monoclonal antibody against human cytomaglovirus (HCMV) was used to detect immediate early antigen (IEA) and early antigen (EA) of HCMV infection in liver tissue of 72 pediatric cases (34 autopsies and 38 biopsies). The HCMV antigen was positive in 25 % (18/27). Among them, 12 cases were both HCMV-IEA and EA positive; 4 were HCMV-IEA positive and 2 HCMV-EA positive only. Liver HCMV infection rate in neonates, the infants with the age <1 year and >1 year was 8. 0 %, 60. 0 %, and 14. 8 %, respectively, indicating that liver HCMV infection occurred at various ages. The liver HCMV infection rate in different diseases was 50. 0 % in infantile hepatitis syndrome; 70. 0 % in extrabiliary malformation, and 12. 5 % in other hepatopathies, suggesting that infantile hepatitis syndrome and extrabiliary malformation were related with HCMV infection in liver tissues.

For rapid diagnosis of enteroviral infection in clinic practice, we developed a reverse transcription and polymerase chain reaction (RT-PCR) assay. Primers homologous to the conserved 5′ non-coding region were designed by analyzing enteroviral genomes, and then they were used to enzymatically amplify RNA from 31 prototype enteroviral strains and enteroviruses (EV) in cerebrospinal fluid (CSF) of 34 cases of aseptic meningitis and 11 cases of aseptic encephalitis. The RT-PCR products generated with these enteroviral primers were analyzed by agar gel electrophoresis and dot blot hybridization analysis. 31 EV strains showed an obvious monoclonal amplification band, and all dot blot hybridization results were positive. Four other viruses and cells cultured were all negative. The study of sensitivity of the RT-PCR showed that amplification production were positive to 10⁻²- 10⁻³ 50% tissue culture infective doses. With this assay, 21(61. 8 %) of 34 aseptic meningitis and 8 (72.7%) of 11 aseptic encephalitis contained EV RNA in CSF samples. Two cases of meningitis and one of encephalitis with EV infection were still positive during convalescence. Our results suggest that this RT-PCR method was a fast, sensitive and specific technique for detection of common EV infection.

Dopamine agonists effectively reduce the secretion of prolactin (PRL) in the great majority of prolactinomas and reduce the bulk of the adenomas, as well as have partial therapeutic effect on some patients with acromegaly. The inhibitory effect of bromocriptine (BC), a dopamine agonist, on growth hormone (GH) and PRL secretion of dispersed cells from the pituitary adenomas of 16 cases of acromegaly, which secret GH and PRL simultaneously, were evaluatedin vitro. The significant inhibitory effects of BC on PRL secretion were found in 12 cases. It was also found that PRL secretion was strongly inhibited when GH was suppressed; on the contrary, when GH secretion was not suppressed, the production of PRL was not or weakly inhibited. The exact mechanism of the effects is unclear so far. It is necessary to investigate, at molecular level, the etiology of GH-PRL adenomas and its response to therapeutic agents.

To study the relationship between constitutive nitric oxide synthase (cNOS) and pregnancy induced hypertension (PIH), cNOS expression and localization in placental villi of PIH patients (n=15) and normal pregnancy patients (n=15) were immunohistochemically studied. The positive immunostaining of cNOS was located in trophocytes and cytoplasm of vascular endothelial cells. The positive rate in PIH patients was much higher than that in cases of normal pregnancy. The positive rates were 34. 40 % in mild, 44. 74 % in middle and 50. 14% in severe PIH patients respectively. There was a significant difference in positive rate among the mild, middle and severe PIH patients (P<0.01). It is concluded that the increase of cNOS activity probably was the results of protective or compensatory mechanism of PIH course.

In order to explore the mechanism of hemodynamic changes caused by high biliary tract pressure, we established an animal model of high biliary tract pressure, in which the disturbance of hemodynamics developed. The cervical or abdominal vague nerve was then blocked. It was observed that when the biliary tract pressure was increased to 16 kPa and kept for 1 h, the arterial blood pressure and cardiac output decreased immediately and parallelly (P<0.05). When the cervical or abdominal vague nerve were blocked or the pressure of the biliary tract was decreased to zero, both indices returned to normal immediately (P> 0. 05). The change of cardiac output lags a little behind that of arterial blood pressure. It suggests that the signal of biliary tract pressure increase can be sent to the cardiovascular center through vague nerve, and the balance between sympathic and parasympathic nerve was broken, which led to the weakening of cardiac contraction and decrease of cardiac output. Due to the peripheral effects of vague nerve, hemodynamic resistance of vessels decreased, which brought about redistribution of peripheral blood flow. Both were the causes of hemodynamic disturbances. After the blood pressure decreased markedly, it showed a jump to normal state when cervical vague nerve was blocked. And the amplitude of diastolic blood pressure restored more than that of systolic blood pressure. This suggests that the cardiac output and peripheral blood resistance are important factors that cause the decrease of blood pressure.

The bacterial inhibitory ability of a new drug delivery system (DDS): Ciprofloxacine/tricalcium phosphate delivery capsule (CTDC), itsin vivo drug release pattern, and the influence of ultrasonic irradiation on its drug release were investigated. It was found that CTDC had a strong and sustained inhibitory ability to some common pathogens of bone and joint infections, such as staphylococcus aureus, escherichia coli and pseudomonas aeruginosa.In vivo drug-release study in animals demonstrated a high concentration of ciprofloxacine in the bone tissue surrounding CTDC which was placed in the greater trochanter of the rabbit and continued to release ciprofloxacine for at least 5 weeks and the blood level of ciprofloxacine was low.In vivo study also showed ultrasonic irradiation could increase the amount of ciprofloxacine released from CTDC, which may be an economical, effecient and safe new method to achieve the control of drug release from DDS.

Presented in this paper are 3 cases of hemorrhage of ascending aorta and left ventricle after open heart surgery treated by extracardial bypass in our hospital from Oct. 1994 to Dec. 1995. Remained aneurysmal wall enclosing conduit graft was used as a sac bypassed to right atrium to form a extracardial left-to-right shunt in order to control bleeding and the results turned out to be satisfactory. The bypass and hemodynamically ignorable shunt can close spontaneously without complications with recovery of coagulation system. The technique may find wide application in clinical practice.

Surgical repair of hypospadias was successfully performed by using free peritoneal graft in the model of rabbit hypospadias. The results showed that free peritoneal graft used as a substitute for urathra had a high survival rate, and the canal was formed well. Our study demonstrated that peritoneum could be used for the surgical repair of hypospadias and other urethral disorders such as urethral stricture.

To evaluate the safety and efficacy of intravaginal misoprostol for cervical ripening in the third trimester, a randomized, double-blind, placebo-controlled trial was conducted in 85 patients indicated for induction of labor and with unfavorable cervices. They were randomly assigned to receive either intravaginal misoprostol (100 mg) or placebo placed in the posterior vaginal fornix. The Bishop score, fetal heart rate and Doppler blood flow velocity waveforms were measured before and 12 h after drug administration. Placenta and decidu were histopathologically observed in some cases. Among 85 patients enrolled, 43 received misoprostol and 42 received placebo. Whereas the mean initial Bishop scores were not significantly different between the two growps, the mean Bishop score in misoprostol group was significantly better than those in placebo group. The mean change in Bishop score was also significantly different (4. 4 for misoprostol versus 1. 0 for placebo,P<0.01). The prevalence of spontaneous onset of labor within 12 h after drug insertion in misoprostol group (67. 4%, 29/43) was significantly higher than that in placebo group (14. 3%, 6/42),P<0.01. The average Doppler velocity systolic to diastolic (S/D) ratios of umbilical artery, middle cranial artery, renal artery were not significantly different before and 12 h after drug insertion between both groups. There was no significant difference in frequency of abnormal fetal heart rate tracings or fetal distress and in the mean Apgar scores between the two groups. Except the presence of vasodilation in villi vessels in the misoprostol group, the placental and decidual histopathological changes had no significantly difference between two groups. It is concluded that intravaginal misoprostol may be an effective and safe cervical ripening agent in the third trimester of pregnancy.

In order to explore the mechanism of anisodamini hydrobromidum (654-2) in treating acute ischemic renal failure, the model of acute ischemic renal failure in white New Zealand rabbits was established to dynamically observe and statistically analyze the intracellular concentration changes of free calcium ( [Ca²⁺]_i) and inositol triphosphate (IP₃). The results showed that the levels of [Ca²⁺]_i and IP₃ in acute renal failure group were higher than those in control group (P<0.01). However, the levels of [Ca²⁺]_i and IP₃ in 654-2 treated group were significantly lower than those in acute renal failure group (P<0.001). It was concluded that 654-2 could alleviate Ca²⁺-overload in renal histocytes in acute ischemic renal failure. The protective mechanism is associated with intracellular reduction of IP₃.

A 61 year-old right handed man, who suffered from right cerebral infarction with evidences of visual-spatial neglect and constructive disorder, was reported. When copying simple geometric designs, he omitted to copy figures on the left side of the page; he tended to bisect the line to the right of the line’s real center; after memorizing the familiar pictures he mainly mentioned the pictures on the right side of the page; when copying the “Rey Complex” he also ignored the structures on the left side. The relations of the neglect and construction disorder are discussed.

Sixty-six cells were examined for chromosome length change, with half of them (33) in the early-metaphase and the other half in the pre-metaphase. The 22 couples of chromosomes (excluding sex chromosomes) in each cell were analyzed. The differences of chromosome length between the two phases were demonstrated and the correlation dimension of each chromosome was calculated. It was suggested that the mitosis of human chromosome possessed the feature of chaotic decrease. This offers a new approach to the research of the dynamic process of the mitosis.