Objective　To investigate the molecular regulation of G1 arrest of mouse thymocytes induced by ionizing radiation. Methods　Cell cycle was analyzed by flow cytometry (FCM) following staining of cells with proidium iodide. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression. Results　It was demonstrated that G1 phase of mouse thymocytes increased significantly at 12h after whole body irradiation (WBI) with the doses of 0.5, 1.0 and 2.0 Gy, and at 24h following 2.0Gy exposure, measured by FCM. In the time course experiment, it was found that G1 phase of thymocytes increased significantly at 4h, reached a peak level at 24h and came down toward 48h after WBI with 2.0Gy X-rays. The results also showed that after 2.0Gy exposure, the expression of proteins in mouse thymocytes increased significantlty from 1h to 8h for p53, for p21 from 4h to 48h, and for MDM2 at 4h and 8h, measured by FCM. But no change was found for GADD45 protein expression. Conclusion　These results suggest that G1 arrest could be induced by a single dose of 0.5 Gy, 1.0Gy or 2.0Gy, and its molecular control might be established through the p53-p21 pathway.

OBJECTIVETo study the role of the HBV-infected mothers' PBMC in intrauterine transmission of HBV to their fetuses.

METHODSThirty pregnant women with serum HBV DNA negative and PBMC HBV DNA positive and their newborns were used as the study group. Ten pregnant women with serum HBV negative and their infants served as the control group. HBV DNA in serum and in PBMC was detected using nested polymerase chain reaction (n-PCR). The mothers' PBMC in newborns' peripheral blood was examined using heminested-PCR.

RESULTSFour newborns were serum HBV DNA positive and 8 newborns were HBV DNA positive in PBMC in the study group. Among them, 2 newborns were HBV DNA positive in both serum and PBMC, 6 cases were positive in PBMC only, and 2 cases were positive in serum only. Five mothers had the GSTM1 gene; and it was not detected in 3 newborns. Among the 8 newborns with HBV DNA positive in PBMC, 3 did not have the GSTM1 gene, at the same time their mothers possessed the GSTM1 gene. Mothers' PBMC were detected in all of these three newborns' peripheral blood. HBV DNA in serum and in PBMC of the control group infants were all negative.

CONCLUSIONHBV-infected PBMC of the mother may serve as a vector in HBV intrauterine infection.

Adult ; DNA, Viral ; analysis ; Female ; Hepatitis B virus ; isolation & purification ; Hepatitis B, Chronic ; blood ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Leukocytes, Mononuclear ; virology ; Pregnancy ; Pregnancy Complications, Infectious ; virology

OBJECTIVETo explore the effect of extract from Chinese chive seed in warming kidney and enhancing yang.

METHODThe influence of extract from Chinese chive seed on the erection of penis of was investigated in adult male rats with experimental insufficiency of the kidney-yang produced by both removal of double spermaries and high dose of hydrocortisone.

RESULTThe extract of Chinese chive seed enhanced the responsiveness of the penis of emasculate rats to outside stimulus, promoted the resistance of the emasculated rats to cold and tiredness and increased autonomous activity.

CONCLUSIONThe extract of Chinese chive seed has the effect of warming kidney and enhancing yang.

Animals ; Chive ; chemistry ; Cold Temperature ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Fatigue ; prevention & control ; Hydrocortisone ; Kidney Diseases ; chemically induced ; physiopathology ; Male ; Mice ; Motor Activity ; drug effects ; Orchiectomy ; Penile Erection ; drug effects ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Seeds ; chemistry ; Yang Deficiency ; chemically induced ; physiopathology

To produce membrane-permeable human epidermal growth factor (hEGF), a pPTD-hEGF prokaryocyte expression vector was constructed and transformed into E. coli BL 21 (DE3). The PTD-hEGF fusion protein was induced by 0.3 mmol/L of IPTG expressed in the form of inclusion body with an yield of 40% of the total protein in the cells, and then purified by Ni2+ -NTA affinity chromatography. The SDS-PAGE analysis showed a single fusion protein band with a molecular weight of 16 kD. The amino acid sequence was checked by MALDI-TOF-MS analysis. The genetic engineering PTD-hEGF fusion protein obviously promoted the proliferation and growth of the HEK-293 cells in vitro.
Amino Acid Sequence ; Cell Line ; Cell Proliferation ; drug effects ; Epidermal Growth Factor ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Transduction, Genetic

OBJECTIVETo identify the mutation of Cu/Zn superoxide dismutase(SOD1) gene in an amyotrophic lateral sclerosis (ALS) family with unique phenotype.

METHODSFive exons of SOD1 gene were amplified by PCR. The differences of these products were analyzed by PCR-single strand conformation polymorphism and visualized by silver staining.

RESULTSAbnormal bands were found in exons 2 and 5 of SOD1 gene in several familial members. DNA sequence analysis verified that a base pair insertion occurred in the codon area of exon 2 and in the intron area of exon 5. And the insertion mutation of exon 2 led to a frameshift mutation and premature stop. It is a new type of SOD1 mutation which may be associated with familial amyotrophic lateral sclerosis.

CONCLUSIONInsertion mutation of exon 2 may be responsible for the disease of an ALS family in Chongqing.

Adult ; Amyotrophic Lateral Sclerosis ; genetics ; Humans ; Mutation ; Polymorphism, Single-Stranded Conformational ; Superoxide Dismutase ; genetics ; Superoxide Dismutase-1

OBJECTIVETo understand the HBV infection rate of peripheral blood mononuclear cells (PBMCs) from fetuses of HBsAg positive mothers, associated risk factors and to explore the clinical significance of detecting HBV infected PBMCs.

METHODSSixty eight pregnant women who were delivered at the First Hospital of Xi'an Jiaotong University, China from August 1995 to February 1997, and their newborns were studied. They were divided into two groups according to their status of HBV serological markers. The study group included 50 cases who were serum HBsAg positive and 18 cases without any HBV serum markers served as control group. All these cases had no symptoms of hepatitis, high risk premature labor, premature delivery and hypertensive disorder complicating pregnancy. Age and gestational age were matched in two groups. Blood samples (5 mL) were taken from the peripheral vein of pregnant women before delivery and from newborns within 24 h after birth, before inoculation of HBV vaccine (HBVac) and injection of hepatitis B immunoglobulin (HBIG). PBMCs were isolated. The sera and PBMCs were stored at -80 degrees C. HBV-DNA in serum and PBMCs were detected with nested polymerase chain reaction (n-PCR). Two pairs of oligonucleotide primers, the outer primer pair for first PCR and inner primer pair for second PCR, designed according to region S of HBV genome were synthesized by Shanghai Cell Biology Institute of Chinese Academy of Science.

RESULTSThe detection rate of HBV-DNA in serum and PBMCs from HBsAg positive pregnant women was 60.0% (30/50) and 40.0% (20/50), respectively. The detection rate of HBV-DNA in serum and PBMCs from newborns of HBsAg positive pregnant women was 46.0% (23/50) and 30.0% (15/50), respectively. Ten newborns were HBV-DNA positive in serum only, 2 were positive in PBMCs only and 13 were positive in both serum and PBMCs. In the control group, HBV-DNA was not detected in PBMC nor in serum. The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of mothers who were HBV-DNA or HBeAg positive in serum (P < 0.05, P < 0.01); the positive rate was significantly higher in the group of mothers who were HBV-DNA positive in both serum and PBMC than that in the group of mothers who were serum HBV-DNA positive only (P < 0.01); and it was markedly higher in the group of mothers who were PBMC HBV-DNA positive than that in group of mothers who were HBV-DNA negative in PBMCs (P < 0.01). The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of newborns who were HBV-DNA positive in serum than that in the group of newborns who were HBV-DNA negative in serum (P < 0.05).

CONCLUSIONSThe positive rate of HBV-DNA in PBMCs from newborns of HBsAg positive pregnant women was 30.0% (15/30). It was related to HBV viremia level and HBV-DNA status in PBMCs of mothers and newborns. Detection of HBV-DNA in PBMCs may be an important supplementary method to determine intrauterine HBV infection, and can predict the response to HBV vaccine.

Adult ; Case-Control Studies ; DNA, Viral ; blood ; Female ; Hepatitis B Vaccines ; administration & dosage ; Hepatitis B virus ; immunology ; isolation & purification ; Humans ; Immunoglobulins ; administration & dosage ; Infant, Newborn ; blood ; Infectious Disease Transmission, Vertical ; prevention & control ; Injections, Intramuscular ; Leukocytes, Mononuclear ; virology ; Male ; Mothers ; Polymerase Chain Reaction ; Pregnancy ; blood ; Risk Factors ; Time Factors ; Treatment Outcome

OBJECTIVETo explore the relationship of aging with the changes of endogenous carbon monoxide (CO), cGMP and cAMP contents in the penile tissues of rats.

METHODSTwenty-four male rats were equally divided into an 8-month, a 16-month and a 24-month group, and their penile erection was detected by injecting apomorphine, their penile cavernous body harvested, and the contents of CO, cAPM and cGMP detected by improved dual wavelength spectrophotometry.

RESULTSThe contents of CO, cAPM and cGMP were reduced with the increase of age, with statistically significant differences between the three age groups (P < 0.01).

CONCLUSIONAging significantly decreased the contents of CO, cAMP and cGMP in the penile tissues of the rats, which suggests that aging might play an important role in erectile dysfunction.

Aging ; physiology ; Animals ; Carbon Monoxide ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Male ; Penis ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of ionizing radiation on the expression of p16, CyclinD1, and CDK4 in mouse thymocytes and splenocytes.

METHODSFluorescent staining and flow cytometry analysis were employed for the measurement of protein expression.

RESULTSIn time course experiments, it was found that the expression of p16 protein was significantly increased at 8, 24, and 48 h for thymocytes (P < 0.05, P < 0.01, and P < 0.05, respectively) and at 24 h for splenocytes (P < 0.05) after whole body irradiation (WBI) with 2.0 Gy X-rays. However, the expression of CDK4 protein was significantly decreased from 8 h to 24 h for thymocytes (P < 0.05-P < 0.01) and from 8 h to 72 h for splenocytes (P < 0.05-P < 0.01). In dose effect experiments, it was found that the expression of p16 protein in thymocytes and splenocytes was significantly increased at 24 h after WBI with 1.0, 2.0, and 4.0 Gy (P < 0.05-P < 0.01), whereas the expression of CDK4 protein was significantly decreased with 2.0 Gy for thymocytes (P < 0.05) and 0.5-6.0 Gy for splenocytes (P < 0.05-P < 0.01). Results also showed that the expression of CyclinD1 protein decreased markedly in both thymocytes and splenocytes after exposure.

CONCLUSIONThe results indicate that the expression of p16 protein in thymocytes and splenocytes can be induced by ionizing radiation, and the p16-CyclinD1/CDK4 pathway may play an important role for G1 arrest of thymocytes induced by X-rays.

Animals ; Cyclin D1 ; biosynthesis ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; Cyclin-Dependent Kinases ; biosynthesis ; Dose-Response Relationship, Radiation ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Proto-Oncogene Proteins ; Radiation Dosage ; Spleen ; cytology ; metabolism ; radiation effects ; Thymus Gland ; cytology ; metabolism ; radiation effects ; X-Rays

Objective To explore the causes,clinical manifestation and therapy of frontal sinusitis after transfrontal craniotomy.Methods Thirty-three patients with frontal sinusitis after transfrontal craniotomy were included in the study.Among them,7 cases had frontal sinus abscess and 4 cases had frontal sinus fistula.Twenty-three patients were treated with traditional frontal sinus surgery with facial incision.The nasofrontal dilatation tube was positioned for more than 3 months.Nine patients were treated with endoscopic frontal sinus surgery,and 1 patient was treated with combined endoscopic and traditional frontal sinus surgery,with nasofrontal dilatation tube positioned for less then 1 month.In the revision surgery,the bone wax and phlogistic acestoma were cleaned out in both operational methods.The causes of frontal sinusitis after transfrontal craniotomy were discussed by studying the frontal sinus CT image,and prior surgical data.Results All patients were followed up for more than 6 months after the nasofrontal dilatational tube was removed.Among 33 patients,two cases with traditional frontal sinus surgery were operated twice due to nasofrontal dilatation tube fall off in 1 month.In all 33 patients,30 cases cured and 3 cases got better.There were no curative difference between two operational methods.Conclusions The causes of frontal sinusitis after transfrontal craniotomy were inadequate sinus management in craniotomy and bone wax tamping in frontal sinus.There was more frontal sinus abscess and fistula occurring in frontal sinusitis after transffontal craniotomy than that in ordinary frontal sinusitis.The therapy included cleaning out bone wax and phlogistic acestoma,and expanding the frontal sinus ostium.The satisfying curative effect was obtained in both operational methods,but endoscopic frontal sinus surgery Was better because it is minimally invasive,no facial incision and quick recovery with less nasofrontal dilatational tube posting time.

OBJECTIVETo study the possible role of JNK1, Raf-1 and Livin in the carcinogenesis of sporadic colorectal tubular adenoma.

METHODSImmunohistochemical staining was used to detect the expression of JNK1, Raf-1 and Livin proteins in 65 sporadic colorectal tubular adenomas with dysplasia of varying degrees and 22 colorectal tubular adenoma with cancerous area.

RESULTSIn normal colorectal mucosa, colorectal tubular adenoma with dysplasia and colorectal tubular adenoma with cancerous area, the positive rate of JNK1, Raf-1 and Livin expression was increased gradually. The positive expression of JNK1, Raf-1 and Livin was all significantly higher in the cases of colorectal tubular adenoma with dysplasia or with cancerous area than that in normal colorectal mucosa (P < 0.05), and the positive expression of JNK1, Raf-1 and Livin was significantly higher in colorectal tubular adenoma with cancerous area than that in colorectal tubular adenoma with dysplasia of different degrees (P < 0.05). In the cases of colorectal tubular adenoma with dysplasia of varying degrees, the positive expression of Raf-1 was increased along with the increasing dysplasia degree of colorectal tubular adenoma (P < 0.05). Coexpression of JNK1, Raf-1 and Livin increased gradually in the carcinogenesis of sporadic colorectal tubular adenoma, while positive correlation was found among the expressions of JNK1, Raf-1 and Livin.

CONCLUSIONJNK1, Raf-1 and Livin may be involved in the carcinogenesis of sporadic colorectal tubular adenoma.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenoma ; metabolism ; pathology ; Adult ; Carcinoma ; metabolism ; pathology ; Cell Transformation, Neoplastic ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Intestinal Mucosa ; metabolism ; pathology ; Male ; Mitogen-Activated Protein Kinase 8 ; metabolism ; Neoplasm Proteins ; metabolism ; Precancerous Conditions ; metabolism ; pathology ; Proto-Oncogene Proteins c-raf ; metabolism