Human calcitonin (hCT) is a 32 amino acid peptide hormone that requires C-terminal amidation for full biological activity. Calcitonin has important physiological function in vivo. We describe the couple expression of a synthesized modified human calcitonin(hmCT) gene fused with glutathione-S-transferase and rat peptidylglycine alpha-amidation monooxygenase (PAM) in insect cells infected by recombinant baculovirus GSTCT/PAM. Using Western blotting against hmCT or rat PAM, the GSThmCT fusion protein had been identified as well as the PAM. Following affinity chromatography with glutathione agarose column, the GSThmCT fusion protein produced by insect cells was purified. The purified fusion protein was also interacted with antibody against hmCT. The couple expression of a modification enzyme and its substrate in eucaryotic expression system may be used for producing other biological activity peptides.
Animals ; Baculoviridae ; genetics ; Calcitonin ; biosynthesis ; genetics ; Cells, Cultured ; Gene Expression ; Glutathione Transferase ; genetics ; Humans ; Insecta ; cytology ; Mixed Function Oxygenases ; biosynthesis ; genetics ; Multienzyme Complexes ; biosynthesis ; genetics ; Rats ; Recombinant Fusion Proteins ; genetics

Hepatocellular carcinoma (HCC) is a common malignant tumor worldwise. The incidence rate of HCC is high and is easy to metastasis and recurrence, which seriously affects human health. Traditional chemical drugs have some challenges such as toxicity, side effects, and multidrug resistance, thus it is urgent to find new drugs and effective targets. Here we synthesized a novel chemical, protonic bis-phenanthroline (H-BP), and the antitumor effect was investigated in the study. The results showed that H-BP could selectively inhibit the proliferation of tumor cells and cause HCC apoptosis. And also, in HCC tumor-bearing mice, H-BP could effectively prevent the growth of tumor mass, even completely eliminate the tumor at medium dose (5 mg·kg^-1) and high dose (10 mg·kg^-1), and meanwhile H-BP has no significant effect on the body weight of mice. The experimental protocol was approved by the Animal Ethics Committee of Southwest University, and the experimental operation was strictly carried out in accordance with the ethical principles of animal use and care. Mechanism studies showed that H-BP induced HCC apoptosis was related to down-regulation the expression of pleomorphic adenoma gene like-2 (PLAGL2), a oncogene transcription factor, resulting in the down-regulation of PLAGL2 downstream proteins hypoxia inducible factor and β-catenin. This study not only introduces the dimerization method to form novel compounds that will provide a new approach for drug design, but also suggests that PLAGL2 may be an effective target in tumor therapy.

UNLABELLEDFrom large-scale sequence of human fetal liver cDNA library, we have obtained a full-length cDNA from an EST after further sequencing. It has been demonstrated by the alignment comparison with data base available that it is a novel member of Ubc family and got the number from GeneBank: UBF-F1 AF 294842.

AIM AND METHODSTo demonstrate its authenticity, UBF was amplified from the total RNA of human fetal liver and HL-60 cell line using RT-PCR, and the PCR products were further sequenced and compared with the original UBF sequence. To evaluate the expression level and subcellular location of UBF in human multiple tissues, in situ hybridization was carried out on the frozen section of human fetal multiple tissues and HL-60 cell line with DIG-labeled UBF cDNA probes.

RESULTSThe experimental results of RT-PCR and sequencing showed that the sequence of RT-PCR products were the same as the original UBF. The experimental results of in situ hybridization showed that UBF was expressed widely by human multiple fetal tissues and the expression level were very high in HL-60 cells.

CONCLUSIONIt is suggested that the special structure of UBF is authentic, and the expression profiling research of UBF shows that UBF is expressed widely by human multiple fetal tissues and the expression level is very high in HL-60 cells, implying that UBF plays the important function in the developing tissues and leukemia cells. It is also suggested that UBF may be functionally related with the nucleic-involving cellular activities based on the results of sub-cellular localizations.

Amino Acid Sequence ; Cloning, Molecular ; Gene Expression Profiling ; HL-60 Cells ; Humans ; Molecular Sequence Data ; Pol1 Transcription Initiation Complex Proteins ; genetics ; Reading Frames ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Ubiquitin-Conjugating Enzymes ; classification ; Ubiquitination

Colectomy ; Colonic Neoplasms/surgery* ; Colonoscopy ; Humans ; Lymph Node Excision ; Lymph Nodes/pathology* ; Robotic Surgical Procedures

Animals ; Apoptosis ; Concanavalin A ; toxicity ; Liver ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Nitric Oxide ; biosynthesis ; Tumor Necrosis Factor-alpha ; biosynthesis

Actins ; analysis ; Animals ; Cell Division ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Estradiol ; pharmacology ; Liver ; cytology ; drug effects ; Liver Cirrhosis ; etiology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo investigate the time trends of cancer mortality among residents in Kaifeng county, Henan province.

METHODSData on cancer mortality from the vital registration system in Kaifeng county from 1988 to 2005 was analyzed. A total of 9543 death records (5974 males and 3567 females) due to malignant tumors were studied. A two-year-period age-specified standardized mortality rates were directly adjusted by the world standard population, and the annual percentage change (APC) of mortality were estimated by a linear logarithm regression.

RESULTSThe crude cancer death rate for male was 95.09/100,000 and its age-standardized death rate was 117.41/100,000. While, the crude cancer death rate for female was 59.13/100,000 and the age-standardized death rate was 57.15/100,000. There was a significant growth tread for lung cancer (APC: 6.54%), liver cancer (5.07%) in males and breast cancer (7.04%) in females in the groups aged over 18. On the contrary, the decreasing treads for esophageal cancer in both of sexes (-7.09%, -13.53%) were also observed in this study. Meanwhile, there was no other significant changes in the trend, either in the tumor sites or mortality, was observed.

CONCLUSIONIn the past two decades, there has been a significant increasing trend for cancer mortality in Kaifeng county, of Henan Province. Hence, it is necessary to enhance epidemiological survey to identify risk factors at the earlier stages.

China ; epidemiology ; Death Certificates ; Female ; Humans ; Male ; Mortality ; trends ; Neoplasms ; mortality ; Rural Population

Aged ; Alcoholic Beverages ; Humans ; Hyponatremia ; etiology ; Male ; Substance Withdrawal Syndrome ; complications

Objective To explore the value of stable micro bubble test (SMT) for early diagnosis of hyaline membrane disease (HMD), and to provide early intervention for clinical diagnosis method of hyaline membrane disease in premature infants. Methods A total of 288 premature infants admitted to neonatal intensive care unit (NICU) of Ankang City Central Hospital from January 1st, 2013 to January 1st, 2016 were involved into the study. The primary gastric juice 1-2 mL was extracted for SMT test within 30 minutes after the birth. The number of the stable micro bubble with diameter<15μm was counted by low magnification. The clinical symptoms and X-ray results within 6 h after the birth were used as the gold standard of diagnosis of HMD. The premature infants were divided into the case group and control group. The general information was compared between two groups. The optimal critical stable micro bubble value was determined by the ROC curve. The number of micro bubbles <5/mm2, <10/mm2, <15/mm2 and <20/mm2 were used as the threshold, and the sensitivity, specificity, positive and negative predictive values were calculated for HMD diagnosis. Results Among the 288 cases of premature infants, 91 cases (31.6%) were diagnosed as HMD, 197 cases (68.4%) were diagnosed as non HMD. There were lower gestational age, lower birth weight and more proportion of male babies in the case group compared with those of control group (P<0.05). There were no significant differences in prenatal usage of hormones, premature rupture of membranes, the proportion of cesarean section and 5-min Apgar scores between two groups. The number of micro bubbles of 15/mm2 was the optimal threshold for diagnosis of HMD, and AUC was 0.80 (0.73-0.84). When the number of micro bubbles of 5/mm2 was used for the diagnosis, 38 cases were in line with the diagnosed as HMD. When the number of micro bubbles of 10-15/mm2 was used for the diagnosis, 16 cases were in line with diagnosed as HMD. The number of micro bubbles of<15/mm2 was used as boundary value of HMD. The sensitivity, specificity, positive predictive value, negative predictive value and Youden index for SMT prediction and early diagnosis of HMD were 89.0%, 91.4%, 82.7%, 94.7%and 80.4%, which can be used as predictive indicators of HMD. Conclusion SMT test is simple, rapid, economic, high sensitivity and good specificity, which can effectively predict and early diagnose HMD, and have high clinical value.

OBJECTIVETo acquire more knowledge about neonatal lupus erythematosus (NLE).

METHODSeven cases with neonatal lupus erythematosus who were seen in this hospital from 1990 to 2009 are reported in this paper and 87 cases reported previously from 1980 to now in China were reviewed. The clinical manifestations, serum autoantibodies, treatment and results of long-term follow-up are analyzed and summarized.

RESULTTotally 94 cases were summarized. Male/female ratio was 48/46; 73 cases had skin rash; 23 had heart abnormality, among whom 13 had cardiac conductive problems including 8 cases of atrioventricular blockage (AVB) (3 degree I, 3 degree II and 2 degree III) and 5 cases of right bundle branch block cases (RBBB). Nine cases had anatomical abnormality including 5 cases of atrial septal defect (ASD), 2 cases of ventricular septal defect (VSD) and 2 cases of atrial enlargements. Forty-four cases had hematological problems including 28 with thrombocytopenia, 11 with leukocytopenia and 34 with anemia. Thirty cases had hepatic abnormality, including 24 liver dysfunction, 22 hepatomegaly, 6 splenomegaly and 3 cholestasis. Three cases had nephropathy; 3 had elevated creatine kinase; 2 had nervous disorder. Among the 94 cases, 86 (91.5%) were positive for anti-SSA, 51 (54.3%) anti-SSB, 16 anti-ds-DNA, 14 anti-U1-RNP, 13 anti-Sm (+), 6 anti-RNP and 4 anti-rRNP(+). Among the corresponding mothers, 39 cases (44.8%) were asymptomatic before pregnancy, 35 had SLE, 5 had SCLE, 3 had Sjogren syndrome, 2 had chilblain, photosensitivity and arthralgia, respectively, 1 had rheumatoid arthritis and 1 had psoriasis. During pregnancy, 8 mothers developed SLE. Totally 48 mothers (51.1%) suffered from LE. Together with 15 mothers who had transient skin rash during the pregnancy, there were 23 mothers (59%) who had new clinical manifestation among the 39 asymptomatic mothers. Twenty NLE cases accepted glucocorticoid treatment, 4 of them were treated with intravenous immunoglobulin. Sixty-eight cases were followed up for up to 12 years, 58 cases were healthy, 5 cases improved and 3 died. Two cases still had grade III AVB without pacemaker.

CONCLUSIONNLE is a rare acquired autoimmune disease. Although nearly half of the mothers were asymptomatic before pregnancy, more than half of them developed LE or other symptoms. The clinical presentations in Chinese cases include a transient rash, cardiac lesion while grade III AVB was rare, hematological changes and liver impairments which were common but not serious. Anti-SSA and/or anti-SSB were the most related autoantibody. Most patients with NLE have relatively good prognosis.

Adult ; Autoantibodies ; blood ; Female ; Follow-Up Studies ; Humans ; Infant, Newborn ; Lupus Erythematosus, Cutaneous ; drug therapy ; pathology ; Lupus Erythematosus, Systemic ; congenital ; diagnosis ; drug therapy ; pathology ; Male ; Mothers ; Pregnancy ; Prognosis ; Retrospective Studies