Health Promotion ; Humans ; Occupational Health

Objective To investigate the role of directly constitutive activation of p38 mitogen-activated protein kinases(p38MAPKs)signaling in ?-globin gene expression and fetal hemoglobin(HbF)induction,and provide direct data for the relationship between phosphorylation of p38 and erythroid differentiation of human K562 erythroleukemia cells.Methods The human K562 erythroleukemia cells were transfected with pCDNA 3.1-MKK3(Glu)and pCDNA 3.1-MKK3(Ala)recombinant plasmids by lipofectamineTM 2000.Then,the stable cell lines overexpressing constitutively active p38 and constitutively inhibitive p38 activation were established by the addition of G418 to select single cell G418-resistant clones and identification with reverse transcriptase-polymerase chain reaction(RT-RCR)and Western blot assays,named K562-MKK3(Glu)and K562-MKK3(Ala)cells,respectively.Furthermore,the direct effects of constitutively active p38 on the ?-globin gene expression and HbF induction were analyzed by RT-PCR and benzidine staining,respectively.Results The results of RT-PCR and Western blot showed that there were no evident changes in the mRNA and protein levels of p38 for various cell models,but compared with K562,K562-vect,and K562-MKK3(Ala)cells,the phosphorylation of p38 and expression of ?-globin levels in K562-MKK3(Glu)cells were significantly up-regulated.The results of benzidine staining displayed that the mean percentages of positive cells stained by benzidine in K562,K562-vect,K562-MKK3(Ala),K562-MKK3(Glu)cells,and K562-MKK3(Glu)cells treated with SB203580 were(3.2?1.4)%,(3.7?1.2)%,(2.8?0.9)%,(32.6?5.3)%,and(7.8? 2.3)%(q = 7.56 P

1)which represented 340 genes and 171 down-regulated(SLR

BACKGROUND: Recent study showed that osteocalcin may elevate Insulin secretion and sensitivity, prevent the fat accumulation, play a role in the metablism of glucose and lipid. Undercarboxylated osteocalcin works as the main role. OBJECTIVE: To investigate the effect of different concentrations of glucose on osteoblast undercarboxylated osteocalcin. METHODS: The rib trabeculae were resected and broken, trypsinizated and washed completely by PBS. Bone surface and non-adhesive floating cells in cleaning fluid were observed with inverted microscope. Rib trabeculae was washed by DMEM culture medium once, and cultured in culture bottle. The culture liquid was replaced by new one once a week. The osteoblast was moved from the scledte a week later. The cells were fused monolayer and could be subcultured 4 to 6 weeks later. The active second or third generation cells were inoculated to 6-pore plate forming 5 groups. Osteoblast were stimulated by 5.6 mmol/L., 7.6 mmol/L, 9.6 mmol/L, 12.6 mmol/L, 20.6 mmol/L glucose medium respectively after the 80% cells were fused, the vitamin K_2 was added into the culture liquid until the concentration of it to be 10~(-5) mol/L. Supernatant was collected after half hour culturing, the undercarboxylated osteocalcin level were detected with RIA test kit, and corrected it as the total the undercarboxylated osteccalcin, calculated the carboxylated incomplete osteocalcin rate. RESULTS AND CONCLUSION: The rate of ostecblast carboxylated incomplete osteocalcin was different under different concentration glucose. The rate of 7.6 mmol/L, 9.6 mmol/L, 20.6 mmol/L concentration glucose groups were higher than that of 5.6 mmol/L glucose group [(0.27±0.02)%, (0.29±0.04)%, (0.12±0.02)%, P < 0.05]. It is indicated that osteoblast could sense the change of glucose concentration by regulating the secretion of the undercarboxylated osteocalcin between the concentration of 5.6mmol/L to 9.6mmol/L, while the carboxylated incomplete osteocalcin decreased as the concentration of glucose increased.

Mitochondria is an important organelle in mammalian cells with multiple functions,such as ener-gy production and cell homeostasis maintaining. It is known that hundreds of diseases are associated with mi-tochondrial defects. The studies show that the exoge-nous mitochondria can directly enter mammalian cells in vitro, and they also can quickly transform into ani-mal tissues by local or intravenous injection. Current-ly, it has raised a new therapeutic strategy for mito-chondrial diseases, called mitotherapy, which trans-plants exogenous functional mitochondria into mito-chondria-defective cells. The mitochondria in recipient cells play their own roles, including energy produc-tion,maintaining free radical balance,and cell viabili-ty recovery. Since there is no effective method for mito-chondria-related diseases up to now, the mitotherapy will provide a new approach for the prevention and treatment of these diseases.

The aim of this study is to construct specific shRNA expressing plasmids, and to observe their effects on H9c2 cardiomyocytes injury induced by hypoxia/reoxygenation (H/R). RIPK1 and RIPK3 are the key kinases mediating the process of necroptosis. Using recombinant DNA technology, we inserted the synthetic shRNA into pSUPER vector to construct RIPK1-shRNA or RIPK3-shRNA plasmid respectively. We transfected H9c2 cardiomyocytes with the two shRNA plasmids respectively, before we treated them with H/R stimulation. Then, we measured the relevant genes and proteins by real-time PCR and Western blot. Meanwhile,we detected the markers of necroptosis and cardiomyocytes injury. The results showed that inhibition of ripk1 or ripk3 gene expression by its specific shRNA might protect the cardiomyocytes injury induced by H/R stimulation.
Animals ; Apoptosis ; Cell Hypoxia ; Cell Line ; Gene Expression ; Myocytes, Cardiac ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; metabolism ; Transfection

Objective To explore the correlation between brainstem auditory evoked potential (BAEP) combined with mesencephalic morphology and the prognosis of coma patients with severe craniocerebral injury.Methods Forty-seven coma patients with severe craniocerebral injury were examined with BAEP and moaitored dynamically in early period of coma,and all these patients got head CT scans and the ratio of anteroposterior diameter and transverse diameter of mesencephalon were measured at the same time.Results There was a significantly negative relationship between the first BAEP classification and GOS score at the end of the follow-up (r =-0.755,P =0.000 ).Among all of the patients,there were 27 patients with the ratio of anteroposterior diameter and tansverse diameter of mesencephalon from 0.9 to 1.1,19 patients (70.4％,19/27) with good prognosis;20 patients with the ratio ＞1.1 or ＜ 0.9,6 patients (30.0％,6/20 ) with good prognosis.The rate of good prognosis between the two had significant difference ( x2 =7.521,P =0.006).The sensitivity and specificity were 73.7％(14/19),82.6％ (19/23) at the first BAEP combined with mesencephalic morphology,88.2％ (15/17),100.0％ (21/21) at the second BAEP combined with mesencephalic morphology.Conclusion BAEP combined with mesencephalic morphology can evaluate effectively the prognosis of coma patients with severe craniocerebral injury.

Objective To observe the impairment of different doses of cyclosporine A (CsA) to the rat myocardial tissue to offer scientific evidence for the long-term safe application of CsA in heart transplantation. Methods Eighty-four female Wistar rats, each weighing of (200 ± 25)g, were randomly divided into 12 groups. On days 7,14,21 after a constant peritoneal injection of CsA(0,5,10,15 mg/kg) and 1 ml physiological saline in control group, the rats were put to death, the rat myocardial tissue taken, to observe the pathologic and structural changes of the tissue cells under light microscope and electron microscope. The contents of rat myocardium tissue malondialdehyde(MDA) and superoxide dismutase(SOD) were measured;cardiomyocyte apoptosis was detected and accounted, apoptosis index(AI) was measured with the method of TUNEL. Results Small dose of CsA(5 mg/kg)had no obvious effects on cardiac tissue, in CsA groups of 10 mg/kg and 15 mg/kg, under the light microscope, there appeared edema, degeneration and necrosis of myocardium, part of cardiac myocyte had different level cavity;under the electron microscope, there appeared mitochondria damage, nucleus shrinkage and chromatic margination, part of cardiac myocyte had focus cavity. There was dilated endoplasic reticulum in the sarcoplasm. The effects of different time and dose on MDA content of rat myocardium tissue had statistical significance (F = 6.37,10.15, both P < 0.05). Interaction between time and dose existed statistical significance (F=7.14, P< 0.05). The MDA contents of CsA group of 10 mg/kg and 15 mg/kg were [(2.29 ± 0.18), (3.10 ± 0.45), (2.57± 0.37)nmol/L] and [(3.09±0.63), (3.32 ±0.52), (3.34 ± 0.29)nmol/L] on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [(1.98 ± 0.20), (2.04 ± 0.52), (1.99 ± 0.26) nmol/L, all P < 0.05], respectively. The effects of different time and dose on SOD activity of rat myocardium tissue had statistical significance(F = 8.43,11.69, both P < 0.05). Interaction between time and dose existed statistical significance(F = 9.86, P < 0.05). The SOD activity of CsA groups of 10 mg/kg and 15 mg/kg were (15.95 ± 1.00), (12.74 ± 1.31), (14.01 ± 0.81)nmol/L and (13.04 ± 1.01), (14.68 ± 0.81), (14.01 ± 0.63)nmol/L on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [(10.38 ± 0.80), (9.73 ± 0.58), (10.20 ± 0.26)nmol/L, all P < 0.05], respectively. Apoptosis nucleus appeared huffy or brown under the light microscope. The effects of different time and dose on AI of rat myocardium tissue had statistical significance (F = 10.02,20.46, both P < 0.05). Interaction between time and dose existed statistical significance (F = 15.73,P < 0.05). The AI of CsA groups of 10 mg/kg and 15 mg/kg were (6.91 ± 0.70)%, (11.10 ± 2.05)%,(19.81 ± 5.00)% and (11.02 ±2.02)%,(15.51 ± 1.31)%,(33.40±6.60)% on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [(4.40 ± 0.13)%, (4.60± 1.20)%, (5.20 ± 1.10), all P < 0.05] and CsA group of 5 mg/kg [(4.60 ± 0.10)%, (5.00±2.11)%, (5.43± 1.11)%, all P < 0.05], respectively. Conclusion Small dose of CsA has no obvious effects on cardiac tissue, but large dosage can induce myocyte apoptosis and damage by causing oxidative stress;after implantation, attention should be paid to cardiac impairment due to constant large dosage of CsA.

Objective:To study the antimalarial mechanism of benflumetol (B). Methods: Flow cytometry (FCM) was used to analyze the effects of B and chloroquine (CQ) on DNA content of Plasmodium berghei and pH value of the lysosome of malarial parasites. Results: DNA content of the plasmodia not treated with any drugs was not changed in 24 hours,while benflumetol could decrease the DNA content: the DNA content began to decrease 2 h after the drug administration and reached the minimum by 16 h, but somewhat increased at 24 h after administration. The pH in the lysosome increased 1 h and restored premedication level 4 h after benflumetol administration. Chloroquine had the same effects on DNA and lysosome pH of malarial parasites.Conclusions: The antimalarial mechanism of benflumetol is directly related to its effect to inhibit the synthesis of DNA.

Objective To explore the effect of neuroendoscopic surgery for the removal of medium and large sized tuberculum sellae meningiomas through supraorbital keyhole approach.Methods A retrospective research was performed on 7 case of patients with tuberculum sellae meningioma who underwent endoscopic surgery through supraorbital keyhole approach.The main performance of patients as tumor diameter were 2.8-4.7 cm and the skin incision located at superciliary aich which size of intra-frontal bone window was 3.5 cm× 2.0 cm.Results Total removal was achieved in 7 cases(simpson Ⅰ grade in 2 patients,sirnpson 1Ⅱ grade in 5 patients).Postoperative,the visual outcomes of eyes were showed improvement in 9 eyes,remained steady in 3 eyes,and deterioration in 2 eyes.All patients were followed up for 6-13 months and no recurrence was found.Conclusion Neuroendoscopic surgery through supraorbital keyhole approach is an effective method for the resection of medium and large sized tuberculum sellae meningiomas.