Suppressor of cytokine sigaling (SOCS) genes encode a family of protein recently identified as negative feedback inhibitors of signaling by eytokine receptors. We have previously shown that endotoxin markedly stimulates SOCS gene expression in rat liver, that correlates with observed resistance to growth hormone-signaling during endotoxemia. The objective of this study was to determine the expression of SOCS genes in state of fasting that have been shown to cause altered responses in pro-inflammatory cytokines and anzbolic hormones. Male Sprague-Dawley rats (～200g) were fasted for 1, 2 or 3 days, or refed for 3 days following a 3-day period of fasting. Liver and muscle mRNAs were determinedby Northern blotting using specific rat cDNA probes cloned in our laboratory. In liver, after a l-day lag period, there was a progressive 2-fold increase in SOCS-3 and 75% decrease in SOCS-2 mRNA afte 2 and 3 of fasting. Both SOCS mRNAs were normalized by 3 days of refeeding. There was no measurable changes in tyrosine phosphorylation of STAT1, STAT3, STAT5a or STAT5b, nor activation of MAP kinases including ERK 1/2, p38, and JUNK 1/2 in liver by 3 days of fasting. In muscle, there was a similar 75% decrease in SOCS-2 mRNA, but no change in SOCS-3 mRNA following 3 days of fasting. These data suggest that malnutrition regulates SOCS-2 and SOCS-3 in a different way, and this regulation is tissue specific. The changes of SOCS mRANs are independent of measurable phosphoryiation of multiple STATs and activation of MAP kinasea. The altered SOCS expressions during fasting may explain the changes of biological effects of multiple cytokines and anabolie hormones in malnutrition states.

Objective To research the protective effect of Ento-I against cerebral ischemia-reperfusion injury in rats, and to evaluate its analgesic and anticoagulating effects in mice. Methods The ischemic model was established with line embolism to block the middle cerebral artery of male rats. The 56 rats were randomly assigned into 7 groups of sham-operation, blank-matrix, normal saline, Ento-I plastic of 3 doses (6.67, 3.33, 1.67 mg/kg), and ozagrel sodium (8.3 mg/kg, ip). The effect of Ento-I plastic on anti-cerebral ischemia was measured by nervous function scores and the areas of cerebral infarction were determined by TTC staining for the calculation of cerebral infarction rates. The analgesic effect of Ento-I plastic was determined with acetic acid-induced twisting experiment. Sixty KM mice were randomly allocated into blank-matrix, aspirin, aspirin-plastic, and Ento-I plastic of 3 doses (5, 10 and 20 mg/kg), the number of mouse twisting were recorded right after intraperitoneal injection of 0.7% acetic acid solution at the time of 1 h after the last administration. Moreover, the anticoagulant activity of Ento-I plastic was tested by glass capillary method. Results The results of acetic acid-induced twisting experiment displayed that Ento-I plastic of all 3 dose groups (5, 10 and 20 mg/kg) could significantly reduce the number of body torsion and increase the inhibitory rates of twisting, compared with that of blank matrix group (the inhibitory rates of twisting for 3 dose groups were 21.79%, 48.89%, and 56.15%, respectively), with dose-response manner. According to the results of glass capillary test, the clotting time of mouse blood could be significantly prolonged by mid- (10 mg/ kg) and low-dose (5 mg/kg) of Ento-I plastic with corresponding clotting time of (155.20±54.19) s and (155.80±73.84) s, compared with normal saline group at (92.10±24.61) and blank-matrix group at (80.40±48.09, P<0.05). The experiment results of the isch emia-reperfusion injury by line embolism method in rats exhibited that Ento-I plastic in mid-dose (3.33 mg/kg) could significantly re duce the neurological scores after 24 h of reperfusion injury, from (2.33±0.52) of normal saline group to (1.00±0.00) of mid-dose group (P<0.01). The results from TTC staining revealed that the cerebral infarction rates of normal saline group and blank- matrix group were (24.89±7.24) % and (27.72±7.89)%, respectively, whereas those of 6.67 mg/kg and 3.33 mg/kg group of Ento-I plastic were (14.01±2.65) % and (14.73±4.94)%, respectively. Compared to the 2 negative-control groups, both the high- and mid-dose of Ento-I plastic could significantly reduce the cerebral infarction rates after ischemic reperfusion injury in rats (P<0.01). Conclusion Ento-I plastic demonstrates strong analgesic and anticoagulant effects, and could substantially reduce the neurological scores and reduce cerebral infarction rates for ischemia-reperfusion injured rats. These are likely to be the mechanism of action for Ento-I plastic realizing its anti-cerebral ischemia effect.

OBJECTIVETo investigate the expression of tenascin-C (Tn-C) in keloid and hyperplastic scar (HS).

METHODSTissue samples were harvested from 10 patients with keloid and 10 with HS (6 - 10 months) and from the skin of 5 adult healthy volunteers. The expression of Tn-C in these samples was determined with immunohistochemistry method.

RESULTSThere was scarce expression of Tn-C in the skin tissue in adult healthy volunteers, and it was only present in the dermal papillae at the dermis epidermis conjunctions and partly in the blood vessels and skin appendages adjacent to the basement membrane. There was enhanced expression of Tn-C in the dermal scar tissue and skin appendages in both keloid and HS, especially in keloid, which exhibited a diffused pattern in the tissue. When compared with that in normal skin, the Tn-C expression in the normal skin adjacent to the keloid was enhanced markedly, but not in the normal skin near HS tissue.

CONCLUSIONThere was increased Tn-C expression in keloid and HS (6 - 10 months).

Cicatrix, Hypertrophic ; metabolism ; Female ; Humans ; Immunohistochemistry ; Keloid ; metabolism ; Male ; Skin ; chemistry ; Tenascin ; analysis

OBJECTIVETo investigate the influence of recombinant human growth hormone (rhGH) on the mortality of the patients with severe burns.

METHODSIn a prospective multi-center randomized clinical trial, 207 adult patients with severe burns were enrolled in the study, and they were randomly divided into treatment (T, with subcutaneous injection of rhGH) and placebo control (C, with subcutaneous injection of same amount of isotonic saline) groups. The mortality, incidence of hyperglycemia and sepsis in the two groups were observed.

RESULTSThe mortality rate in T group was 0.89% as compared with 5.26% in the C group (P >0.05). Hyperglycemia (blood glucose level over 10 mmol/L) was present in 36.61% of patients in T group but 18.95% in C group (P <0.01). There was no difference in the incidence of sepsis between the two groups (P > 0.05).

CONCLUSIONThe application of rhGH in appropriate dosage in adult patients with severe burns could be safe, but blood glucose level should be monitored during the administration.

Adolescent ; Adult ; Aged ; Blood Glucose ; Burns ; diagnosis ; drug therapy ; Female ; Human Growth Hormone ; therapeutic use ; Humans ; Male ; Middle Aged ; Pregnancy ; Prognosis ; Prospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the association between polymorphisms at -14 bp and zinc finger protein(ZNF) sites of ATP-binding cassette transporter A1 (ABCA1) gene promotor and high density lipoprotein-cholesterol (HDL-C) level and coronary heart disease (CHD).

METHODSPolymorphisms of Bme13901 restriction site at -14 bp and an insertion/deletion site of ACCCC in variable number of tandem repeats-zinc finger protein(VNTR-ZNF) of ABCA1 gene were detected using PCR in 260 CHD patients and 220 healthy subjects from a Chinese population in Tianjin.

RESULTSCT genotype was most common in both groups with no differences found in between (P> 0.05). No differences were found in the frequencies of the rare T allele for -14 bp (P> 0.05). For the -14 bp site, subjects with CT/TT genotype had a lower serum mean concentration of HDL-C compared with those with the CC genotype (P< 0.05). Genotypic frequencies of VNTR-ZNF were 6.2% for the inserted form, 43.8% for the deleted form and 50.0% for the inserted/deleted form. No significant difference was found in the distribution of allele and genotype, or in the levels of HDL-C between the two groups (P> 0.05).

CONCLUSIONThe genotypes at -14 bp of ABCA1 gene are associated with the plasma level of HDL-C. HDL-C levels in T allele carriers were significantly lower (P< 0.05). No association was found between variations in ABCA1 VNTR-ZNF and plasma levels of HDL-C, or between the ABCA1 -14 bp and VNTR-ZNF polymorphisms and susceptibility for CHD.

ATP Binding Cassette Transporter 1 ; ATP-Binding Cassette Transporters ; genetics ; Cardiovascular Diseases ; genetics ; metabolism ; Case-Control Studies ; Cholesterol, HDL ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Promoter Regions, Genetic

A xylosidase gene, labeled as BH1068 in genome of Bacillus halodurans C-125, was successfully cloned and overexpressed in Escherichia coli JM109. The purified enzyme was thoroughly characterized and its xylosidase function was unambiguously confirmed. It has maximum activities in neutral condition and is stable over a wide range of pH (4.5-9.0). The enzyme has a broad temperature optimal (35 degrees C-45 degrees C) and is quite stable at temperature up to 45 degrees C. The unique pH and temperature profiles of the enzyme should allow a wide range of xylanolytic operational conditions. With high specific activity of 174 mU/mg protein for its artificial substrate (p-nitrophenyl-beta-xylose) and low xylose inhibition (inhibitor constant Ki = 300 mmol/L), this enzyme is among the most active and high tolerant bacterial xylosidase to xylose inhibition. Its high synergy with commercial xylanase has been demonstrated with beechwood xylan hydrolysis, achieving a hydrolysis yield of 40%. Its neutral pH optimal and high tolerance to product inhibition complements well with its fungal counterparts that are only optimal at acidic pH and susceptible to xylose inhibition. In conclusion, this enzyme has high potential in the saccharification of xylan and xylan-containing polysaccharides.
Amino Acid Sequence ; Bacillus ; classification ; enzymology ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hydrolysis ; Molecular Sequence Data ; Recombinant Proteins ; genetics ; metabolism ; Substrate Specificity ; Xylose ; metabolism ; Xylosidases ; genetics ; metabolism

Objective    To explore the potential role of tumor spread through air spaces (STAS) as a prognostic indicator of non-small cell lung cancer (NSCLC) through meta-analysis. Methods    PubMed, EMbase and Web of Science, from inception to February 2022 were searched by computer about the research of the 5-year overall survival (OS) and recurrence free survival (RFS) of NSCLC patients with or without STAS. The Newcastle-Ottawa scale (NOS) was used to evaluate the quality of each study. Results    Totally 13 published articles were included with 4 647 patients, and 1 424 (30.6%) patients had STAS. The NOS score of all studies≥6 points. The meta-analysis showed that compared with the NSCLC patients without STAS, those with STAS had a worse prognosis of 5-year RFS, and the combined HR was 1.89 (95%CI 1.61-2.23); they had a shorter 5-year OS, and the combined HR was 2.25 (95%CI 1.79-2.84). There was no statistical heterogeneity among studies. Conclusion    The presence of STAS may be a poor prognostic factor for patients with NSCLC, and enough attention should be paid. The STAS should be recorded in the pathological report to guide the comprehensive treatment and evaluate the prognosis of patients.

OBJECTIVETo construct pGL2-ApoA I luciferase reporter vector containing ApoA I gene regulation area, and to investigate the effect of G --> A and C --> T substitution in ApoA I promoter -75 bp and intron 1 +83 bp region respectively on ApoA I gene expression.

METHODSHuman chromosome DNA fragments containing ApoA I gene were amplified by PCR, and the DNA fragments consisting of ApoA I AA/CC, GG/TT and GG/CC genetypes were selected separately, then pUC vector including above three different DNA fragments was constructed. After digesting pUC vector with Sac I and Bgl II, ligate the different DNA fragments to basic pGL2 vector that containing luciferase reporter gene. Recombinant and PRL-null vector were cotransfected into HepG2 cells by using cationic liposome method. Cells were cultured for 48 h, activity of firefly and renills luciferase was measured.

RESULTSThree vectors with pGL2-ApoA I-L(-2500 to +289 bp) long fragment vectors and 3 with pGL2-ApoA I-S(-145 to +289 bp) short fragment vectors were combinated successfully. Relative activity of luciferase for ApoA I AA/CC or GG/TT was lower than that for GG/CC significantly.

CONCLUSION-75 bp G --> A and +83 bp C --> T substitution in ApoA I gene may inhibit ApoA I gene transcription and expression. It may be the reason why subjects containing -75 bp A and +83 bp T have lower high density lipoprotein cholesterol (HDL-C) concentration.

Apolipoprotein A-I ; genetics ; metabolism ; Cell Line, Tumor ; Cholesterol, HDL ; metabolism ; Genetic Vectors ; Humans ; Introns ; Luciferases ; genetics ; metabolism ; Point Mutation ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Adult ; Cleidocranial Dysplasia ; genetics ; pathology ; Female ; Humans ; Male ; Middle Aged