Objective: To verify whether there exists melatoni n(Mel) receptor in human embryonic nervous system. Methods: Spec ific binding of Mel to embryonic brain and spinal cord was measured by radioliga nd binding assay. Results: 125　I-Mel binding s ites in optomeninx was the most, in eptochiasm and sniff ball was next; GTPγS d ose-de pendently inhibited the binding. Conclusion: The results demonst rate the presence of specific binding of Mel in human embryonic brain and spinal cord. GTPγS has some effect on 125　I-Mel specific binding,support ing the theory that Mel receptor is coupled to inhibitory G-proteins.

Aged ; Female ; Hodgkin Disease ; diagnosis ; pathology ; Humans ; Reed-Sternberg Cells ; pathology

To explore the effect and mechanism of Guhong injection against cerebral ischemia reperfusion injury, SD rats were randomly divided into the sham-operated group, the model group, the nimodipine group, and high, medium and low-dose Guhong injection groups, with 10 rats in each group. The middle cerebral artery embolization (MCAO) model was established to observe neurological deficit symptoms, infarct volume, SOD activity, MDA content, GSH-Px and CAT activity in rats, as well as the contents of t-PA, PAI, TXB2 and 6-keto-PGF1α in serum. The results showed that Guhong injection could obviously promote the recovery of neurological deficit symptoms, narrow the brain infarct volume in rats after surgery, significantlyincrease the activities of SOD, GSH-Px and CAT and decrease the content of MDA. Meanwhile, it also could obviously increase the contents of t-PA and 6-keto-PGF1α and decrease the contents of PAI and TXB2 in serum, indicating that Guhong injection have better antioxidant and antithrombus effects, as well as a significant protective effect against cerebral ischemia reperfusion injury in rats.
Animals ; Antioxidants ; pharmacology ; Brain Ischemia ; drug therapy ; Catalase ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Injections ; Male ; Malondialdehyde ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Superoxide Dismutase ; metabolism

Objective: To determine protein binding characteri stic and signal transmission pathway of melatonin（Mel） receptor(MR) in human e mbryonic peripheral organ tissues. Methods: MR was measured by radio ligand-binding assay and the effect of GTPγS on melatonin specific bindi ng was studied. Results: Mel specific binding sites were det ermined in 16 kinds of human embryonic tissue and this binding could be inhibit ed by GTPγS, supporting the theory that MR is coupled to inhibitory G-proteins system. Conclusion: MR is measured in human embryo tissue, the se results provide experimental data for elucidating the mechanism of the effect of Mel.

Adult ; Carcinoma ; virology ; Cervical Intraepithelial Neoplasia ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; In Situ Hybridization ; Middle Aged ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; virology

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection

The aim of this study is to investigate the critical factor affecting the properties of PLGA microspheres fabricated by a solid-in-oil-in-water (S/O/W) emulsion technique with BSA as a model protein. Prior to encapsulation, the BSA microparticles were fabricated by a modified freezing-induced phase separation method. The microparticles were subsequently encapsulated into PLGA microspheres by S/O/W emulsion method, then Motic BA200 biological microscope, confocal laser scanning microscope, scanning electron microscope were used to observe the structure of S/O/W emulsion and PLGA microspheres. The protein content extracted or released from BSA microspheres was measured by Bradford protein assay method. It was found that NaCl added in the outer aqueous phase effectively suppressed material exchange between the inner and outer phase of S/O/W emulsion. Then, the structure and permeability of obtained microspheres were influenced. As a result, with the increase of NaCl concentration in the outer aqueous phase, the encapsulation efficiency of microspheres significantly increased from 60% to more than 85%, the burst release of microspheres reduced from 70% to 20%, and the particle size decreased from 103 microm to 62 microm. Furthermore, the rehydration of encapsulated protein was also retarded and then integrity of BSA was successfully protected during encapsulation process. In vitro release test showed that BSA released from PLGA microspheres in a sustained manner for more than 30 days.
Delayed-Action Preparations ; Drug Compounding ; Emulsions ; chemistry ; Lactic Acid ; administration & dosage ; chemistry ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Microspheres ; Oils ; Particle Size ; Polyglycolic Acid ; administration & dosage ; chemistry ; Serum Albumin, Bovine ; administration & dosage ; chemistry ; Sodium Chloride ; chemistry ; Water

OBJECTIVELeukemia is a major cause of death of children in China, which accounts for 50 % of all cancers of children. Data from Tianjin Cancer Hospital was analyzed for mortality of leukemia in children under 20 years from 1981 to 2000 in the city of Tianjin.

METHODSAll physicians and medical staff of the hospitals and clinics in the registry area were responsible for filling out the report forms for every new case diagnosed as malignant tumors. Death certificates for malignant tumors have been registered at the local police station and the residential files were checked. All cancer cases with insufficient information were traced to his/her family and relevant persons worked in the clinic. Tianjin Cancer Registry Center periodically conducted an active re-checking program to review all patient records on cancers that was not registered in this period. Tumors diagnosed in this study were coded according to the International Classification of Diseases for Oncology (ICD-O). Mortality rates were calculated by age, sex and date of death.

RESULTSThe types of acute lymphoid leukemia, acute myeloid leukemia and chronic myeloid leukemia were the most common types of childhood leukemia in Tianjin, comprised 69.3%, 20.9 % and 8.0%, respectively. The mortality for childhood leukemia decreased slowly during the period of 1981 to 2000 in Tianjin. Mortality and morbidity ratios were 0.51.

CONCLUSIONCombined with characteristics of individual forms of childhood leukemia mortality, further epidemiological research is needed to prevent childhood leukemia.

Adolescent ; Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Leukemia ; mortality ; Young Adult

China ; epidemiology ; Colonic Neoplasms ; epidemiology ; mortality ; Female ; Humans ; Incidence ; Male

OBJECTIVETo explore the secular trend in incidence and mortality rate of gastric cancer in Tianjin and to provide evidence and reference for making prevention and control strategy for gastric cancer.

METHODSData derived from Tianjin cancer registry system were analyzed by descriptive epidemiological method and Joinpoint model. A total of 17990 gastric cancer cases reported in Tianjin from 1981 to 2002, including 12755 males and 5235 females were studied.

RESULTSThe annual percent change (APC) of crude incidence rate for males and females was -0.92% (Z = -3.85, P = 0.001) and -0.79% (Z = -2.67, P = 0.015), while the APC of standard incidence rate was -3.55% (Z = -13.52, P = 0.000) and -3.47% (Z = -12.85, P = 0.000). There was a descending trend of incidence rate in males and females above 45-years-old, however, in male under 45 years it showed an increased trend and in females it kept stable. The APC of crude mortality rate was -1.66% ( Z = -5.79, P = 0.000) for males and -1.84% (Z = -6.02,P = 0.000) for females, while the APC of standard mortality rate was -4.60% ( Z = -15.79, P = 0.000) for females and -5.36% ( Z = -8.28, P = 0.000) for males during 1989-2002. Mortality and incidence ratio also indicated a downward trend.

CONCLUSIONDespite its declining trend in Tianjin, gastric cancer still remains an important public health problem in facing of the aging society and many risk factors.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Incidence ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Risk Factors ; Stomach Neoplasms ; epidemiology ; mortality ; Survival Rate