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Objective: To analyze the changing trend and characteristics of lymphocyte-platelets ratio (LPR) of early stage in patients with extensive burns, and to explore the prognostic significance of LPR. Methods: A retrospective case series study was conducted. From January 2008 to December 2018, 244 patients with extensive burns were admitted to the First Affiliated Hospital of Naval Medical University, including 181 males and 63 females, aged (44±16) years. The total burned area of patients was 60.0% (42.0%, 85.0%) total body surface area. Platelet and lymphocyte test results of patients were collected on the 1st, 2nd and 3rd day after admission, and LPR of patients was calculated to analyze the changing trend of the three days after admission. Univariate and multivariate logistic regression analysis were conducted to investigate the risk factors or independent risk factors for death of patients, including age, sex, total burn area, area of full-thickness burns and above, inhalation injury, and LPR. According to the 1st day's LPR after admission of patients, the receiver operating characteristic (ROC) curve predicting death of patients was drawn to find the optimal value of LPR. Patients were divided into high LPR group (n=136) and low LPR group (n=108) based on the optimal value of LPR, and the clinical data of total burn area, area of full-thickness burns and above, inhalation injury, tracheotomy, offline time of patients within 28 days, and mortality in the 2 groups were compared. The surviving curve of patients was drawn by Kaplan-Meier method to predict the difference of the 90-day survival rate between the two groups of patients. Data were statistically analyzed with Student's t test, Mann-Whitney U test, and chi-square test. Results: Within 3 days of admission, the LPR of patients showed a time-dependent upward trend. LPR of patients on the 2nd and 3rd day after admission was 8.6 (5.3, 14.4) and 8.6 (4.9, 13.7), respectively, which were significantly higher than the 1st day's 6.3 (4.2, 9.8), with Z values of -4.25 and -3.43, respectively, P<0.01. Univariate logistic regression analysis showed that age, total burn area, area of full-thickness burns and above, inhalation injury, and LPR were all risk factors for death of patients (with odds ratios of 1.03, 1.73, 1.31, 4.74, and 3.11, respectively, 95% confidence intervals of 1.01-1.06, 1.40-2.13, 1.21-1.42, 1.62-13.86, and 1.41-6.88, respectively, P<0.01). Multivariate logistic regression analysis showed that age, area of full-thickness burns and above, and LPR were independent risk factors for death of patients (with odds ratios of 1.06, 1.36, and 2.85, respectively, 95% confidence intervals of 1.03-1.09, 1.19-1.55, 1.02-7.97, P<0.05 or P<0.01). The area under ROC curve of the 1st day's LPR, predicting death of patients, was 0.61 (with 95% confidence interval of 0.51-0.71, P<0.05), and the optimal predicted value was 5.8 with corresponding sensitivity of 77% and specificity of 52% respectively. The total burn area, area of full-thickness burns and above, rates of incidence of inhalation injury, tracheotomy, and mortality of patients in high LPR group were significantly higher than those in low LPR group (with Z values of -3.06 and -3.19, χ2 values of 5.42, 11.64, and 8.45, respectively, P<0.05 or P<0.01). The offline time of patients within 28 days in high LPR group was significantly shorter than that in low LPR group (Z=-2.98, P<0.01). Kaplan-Meier survival analysis showed that the 90-day survival rate of admission of patients in low LPR group was significantly higher than that of patients in high LPR group (χ2=8.24, P<0.01). Conclusions: The early LPR of patients with extensive burns showed a time-dependent upward trend. The LPR on the first day after admission that is closely correlated with total burn area, area of full-thickness and deeper burns, inhalation injury, tracheotomy, and mortality of patients, is an independent risk factor for the prognosis of patients with extensive burns. The first day's LPR after admission is significantly correlated with the 90-day survival rate of patients, which can be used as an evaluation index for the severity of extensive burns.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Blood Platelets , Burns , Lymphocytes , Prognosis , ROC Curve , Retrospective StudiesABSTRACT
OBJECTIVES@#To study the effects of 17β-estradiol (E2) on the regulation of the proliferation of condylar chondrocytes and provide a preliminary discussion on the role of phosphorylate-mammalian target of rapamycin (p-mTOR) in this regulatory process.@*METHODS@#Condylar chondrocytes were isolated from 6-week-old female rats for primary culture. Drug treatment with different concentrations of E2 and/or rapamycin (RAPA) was carried out on second-generation cells. Cell Counting Kit 8 was used to measure the cell viability of condylar chondrocytes after culture for 24, 48, or 72 h, and reverse transcription-polymerase chain reaction (RT-PCR) was applied to detect the relative gene expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), collagen type Ⅱ (COLⅡ), autophagy-related gene 6 (Beclin-1), and autophagy-related gene 5 (ATG-5). Western blot was employed to determine the relative protein expression of ERα, ERβ, Beclin-1, lipid-modified light chain 3B (LC3-Ⅱ), and p-mTOR.@*RESULTS@#E2 could significantly promote the proliferation of chondrocytes cultured @*CONCLUSIONS@#At a concentration of 10
Subject(s)
Animals , Female , Rats , Autophagy , Cell Proliferation , Chondrocytes , Estradiol , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta , PhosphorylationABSTRACT
Objective To examine the effect of miR-301b in the regulating the differentiation of mesenchymal stem cells into adipocytes.Methods Murine ST2 stromal cells were isolated,cultured and induced with adipogenic agents.The expression of miR-301b was detected by qRT-PCR in adipogenic differentiation group and control group.Stromal ST2 cells were transfected with miR-301b mimics followed by adipogenic treatment.qRT-PCR and Western blotting were conducted to detect the changes of adipocyte specific genes and protein expression levels in the miR-301b mimics transfection group and negative control(NC)transfection group. Results qRT-PCR showed that the expression of miR-301b was decreased after adipogenic treatment in ST2 cells.The relative expression level of miR-301b was less in the adipogenic differentiation group (0.219 ± 0.021) than that of the control group (1.000 ± 0.425, P<0.05). The relative expressions of peroxisome proliferator activated receptor γ(PPARγ),CCAAT/enhancer-binding protein α(C/EBPα)and adipocyte fatty acid binding proteins(aP2)were lower in the miR-301b mimics transfecting group than those in the NC transfecting group(P<0.05).The protein levels of the marker gene aP2 and transcription factors PPARγ and C/EBPα decreased in miR-310b mimics transfecting group compared with those of the NC transfecting group (P<0.05). Conclusion miR-301b can reduce adipocyte differentiation.
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ObjectiveTripartite motif 33 (Trim33) is known to play a very important part in regulating osteoblast differentiation, but its role in adipocyte differentiation is rarely reported. The aim of this study was to investigate the regulatory effect of Trim33 on adipocyte differentiation.MethodsBone marrow stromal cell ST2 cells transfected with the Trim33-pcDNA3.1 plasmid were included in the experimental group and those transfected with the pcDNA3.1 plasmid taken as the control. The cells of both groups were treated with adipogenic medium to induce adipocyte differentiation, followed by determination of the expressions of the adipocyte-specific genes CCAAT enhancer binding protein alpha (C/EBPα), peroxisome proliferator-activated receptor gamma (PPARγ), adipocyte characterizing factor FABP4, and adipocytokines adipsin by oil red O staining, qRT-PCR and Western blotting.ResultsThe expression of Trim33 was significantly upregulated in the experimental group as compared with that in the control (88.51±14.31 vs 1.00±0.31, P<0.01). After 5 days of adipogenesis induction, there were dramatically more lipid droplets in the ST2 cells and the A value was markedly higher in the former than in the latter group (0.69±0.03 vs 0.34±0.03, P<0.01). Compared with the control, the cells in the experimental group exhibited remarkable increases in the relative mRNA expressions of C/EBPα, PPARγ, FABP4 and adipsin as well as the protein expressions of Trim33, PPARγ, C/EBPα and FABP4.ConclusionTrim33 promotes lipid accumulation and upregulates the expressions of adipocyte-specific genes in bone marrow stromal cells.
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Objective: To establish an inductively coupled plasma mass spectrometry (ICP-MS) with microwave digestion method for the determination of 32 kinds of inorganic elements in Qingxue Bawei Tablets (QBT), including Be, B, Na, Mg, Al, P, K, Ca, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Mo, Ag, Cd, Sn, Sb, Cs, Ba, Hg, Tl, and Pb, in order to establish the fingerprint chromatogram of inorganic elements and analyze characteristic elements. Methods: The samples with nitric acid and hydrogen peroxide system as digestion reagents were digested via microwave, calibrated by internal standard elements, such as 6Li, Sc, Y, Rh, In, Ho, Bi, and Th. The contents of 32 inorganic elements were analyzed by ICP-MS. The content distribution curve of inorganic elements was plotted. And the principal components were analyzed with the SPSS 19.0 software. Results: The 32 inorganic elements showed good linearity in the selected concentration ranges, with the correlation coefficients over 0.999 5. The detection limits of the 32 elements were in the range of 0.001-6.390 μg/L. The RSD values of precision, stability, and repeatability all met the demands of quantitative analysis. The recovery was 94.36%-105.47%, while their RSD was 1.53%-4.56%. The fingerprint chromatogram was established by the content distribution curve of inorganic elements. Different batches of samples were similar peak shape, and the amount of inorganic elements content in different order tend to be consistent. The results of principal component analysis showed that Co, As, Ba, Ca, Sb, Ti, Ni, Fe, Cs, Se, P, Al, and K might be the characteristic elements in QBT. The content of five heavy metals (Pb, Cd, As, Hg, and Cu) was under the limit requirements of Chinese Pharmacopoeia (2015). Conclusion: The method is simple, rapid, and accurate, and it can be used for the content determination of inorganic elements in QBT. The analysis of inorganic elements can provide certain reference for the quality control, safety evaluation, and clinical application of QBT.
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<p><b>OBJECTIVE</b>To study the role of S6K1 in the induction of SREBP1c in mouse hepatic cell by high glucose stimulation.</p><p><b>METHODS</b>S6K1 shRNA recombinant adenovirus (S6K1Ax) was injected into tail vein of db/db mice and then hepatic triglycerol content was analyzed. Liver specimen were stained with HE. After transfection with S6K1Ax or pU6Ax, mouse hepatic AML12 cells were treated with high glucose, insulin or glucose and insulin, the expression of mSREBP1c was detected by RT-PCR. S6K1 protein was detected by Western blot.</p><p><b>RESULTS</b>Hepatic S6K1 protein in db/db mice was inhibited a week after S6K1Ax injection. Compared with the control group, hepatic triglycerol content of S6K1Ax group was decreased (0.65+/-0.02) mmol/L vs (0.56+/-0.01) mmol/L (t = 4.312, P less than 0.01), hepatocyte fat droplet and vaculor generation were also decreased, fatty liver was improved. The mSREBP1c expression in S6K1Ax transfected cells was lower than that in the control cells (0.03+/-0.01 vs 0.06+/-0.01, t = 5.624, P less than 0.01). Compared with the basal state, SREBP1c expression of both groups was increased on the insulin stimulation, S6K1Ax group was 0.06+/-0.02 (t = 8.452, P less than 0.01) and control group was 0.08+/-0.02 (t = 3.591, P less than 0.05). There is no difference between control and S6K1Ax group by glucose addition (P more than 0.05).</p><p><b>CONCLUSION</b>S6K1 acts on fatty synthesis by regulating mSREBP1c expression.</p>
Subject(s)
Animals , Mice , Adenoviridae , Genetics , Cell Line , Fatty Acid Synthases , Genetics , Metabolism , Gene Expression Regulation , Glucose , Insulin , Liver , Metabolism , Pathology , Liver Cirrhosis , Metabolism , Pathology , Mice, Inbred Strains , RNA Interference , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Ribosomal Protein S6 Kinases, 90-kDa , Genetics , Metabolism , Staining and Labeling , Sterol Regulatory Element Binding Protein 1 , Genetics , Metabolism , Transfection , Triglycerides , MetabolismABSTRACT
Objective To observe expression of neuron specific enolase (NSE) gene in the brain of rats induced by iodine excess. Methods One hundred and fifty one-month weaning Wistar rats were divided into ten groups according to 5 × 2 factorial experiment. Rats were fed with normal feedstuff and water of a series of iodine concentration by adding potassium iodide respectively: norrmal iodine (NI), five-fold high iodine (5HI), ten-fold high iodine(10HI), fifty-fold high iodine(50HI) and one hundred-fold iodine(100HI). After these rats were fed for three or six months, rat serum thyroid hormones were measured by radioimmunoassay including TT4, TT3, FT4, FT3, rT3 and the mRNA level of NSE in rat brain tissue was studied using RT-PCR technique. Results The levels of serum TT4 and TT3 were significantly different in five iodine level groups(F values were 18.867,27.287, both P < 0.01). The interaction between time and iodine level in TT4 was significant in our study(F values were 2.486, P < 0.05). The levels of TT4 and TT3 of 100 HI group at third and sixth month were lower than those of NI, 5HI, 10HI, 50HI groups in the same period (all P < 0.01). The levels of serum FT4, FT3 and rT3 were significantly different at different time(F values were 4.968,27.046,59.776 respectively, P < 0.05 or < 0.01) and in different iodine level groups(F values were 33.058,28.420,17.482 respectively, all P < 0.01). Moreover, the interaction between time and iodine level in FT3 and rT3 was significant in our study(F values were 6.894,5.233 respectively, both P < 0.01). FT4, FT3 and rT3 in 100HI group were lower than that of other iodine dosage groups at the same time (P < 0.05 or < 0.01). The levels of NSE mRNA in brain tissue was significantly different in five iodine level groups (F values were 29.006, P < 0.05). The levels of mRNA NSE of 100HI group in both three and six months (0.61 ± 0.19,0.61 ± 0.22) were all lower than that of any other groups[NI(0.73±0.13 and 0.72 ±0.26), 5HI (0.72 ± 0.15 and 0.72±0.16), 10HI (0.73 ±0.32 and 0.70±0.13), 50HI(0.71±0.18 and 0.69±0.31), all P < 0.05]. The results of correlation analyses show that the levels of serum FT3 and FT4 had correlations with the levels of NSE mRNA (P < 0.05) both in three and six months(r values were 0.987, 0.969 in three month, and 0.890, 0.910 in six month respectively). Conclusions The expression of NSE gene can tolerant the excess of iodine to a certain extent. Exposure to heavy excess iodine(100HI) can decrease the mRNA level of NSE gene. FT4 and FT3 may both have important roles on the regulation of NSE mRNA induced by excess iodine.
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<p><b>OBJECTIVE</b>To observe the effect of overdose iodine on the expression of CCK gene in brains of rats and identify the possible mechanisms.</p><p><b>METHODS</b>One-month weaning Wistar rats were randomly divided into five groups which were fed with normal feedstuff and water supplemented with different concentrations of potassium iodide, named A group (iodine ration was about 6.15 microg per day), B group (iodine ration was about 30.75 microg per day), C group (iodine ration was about 61.5 microg per day), D group (iodine ration was about 307.5 microg per day) and E group (iodine ration was about 615 microg per day). Rats were sacrificed after being fed for three or six months. Then serum thyroid hormones were measured by radioimmunoassay and the mRNA level of CCK gene was studied by using RT-PCR technique.</p><p><b>RESULTS</b>At the end of three months, the values of thyroid hormones in E group [TT4 (45.2 +/- 13.7) nmol/L, TI'3 (0.65 +/- 0.20) nmol/L, FT3 (0.93 +/- 0.45) pmol/L, FT4 (7.07 +/- 2.43) pmol/L, rT3 (0.15 +/- 0.04) nmol/L] were all lower than those in A group [TT4 (76.0 +/- 18.8) nmol/L, TT3 (1.34 +/- 0.41) nmol/L, FT3 (2.45 +/- 0.62) pmol/L, FT4 (15.12 +/- 3.40) pmol/L, rT3 (0.24 +/- 0.04) nmol/L]. There were significant differences between E group and A group on the levels of serum TH (F values are 14.68, 16.03, 21.16, 20.25, 13.52 respectively, P < 0.01); FT3 levels in C and D groups were significantly decreased as compared to A and B groups (F = 21.16, P < 0.05). rT3 level in D group was significantly decreased compared with A,B and C groups (F = 13.52, P < 0.05). At the end of six months, the levels of serum TH in E group (TT4 (51.84 +/- 15.83) nmol/L, TT3 (0.77 +/- 0.22) nmol/L, FT4 (6.88 +/- 2.23) pmol/L, FT3 (0.74 +/- 0.28) pmol/L, rT3 (0.14 +/- 0.03) nmol/L) were lower than those in any other groups (F values were 6.05, 12.22, 11.25, 13.42, 5.89 respectively, P < 0.05). At the end of both three and six months, the mRNA levels of CCK gene in E group were lower than any other groups (F values were 4.04, 3.95 respectively, P < 0.01). The results of correlation analysis showed that serum FT4 had linear correlation with levels of CCK mRNA (r values were 0.990, 0.948 respectively; P < 0.05); However serum FT3 had no linear correlation with the levels of CCK mRNA (r values are 0.970, 0.932 respectively).</p><p><b>CONCLUSIONS</b>Exposure to overdose of iodine (iodine ration was 100-fold higher than that of A group) could decrease the mRNA level of CCK gene. Compared with FT3, FT4 might have more important role on the regulation of CCK mRNA induced by excess of iodine.</p>
Subject(s)
Animals , Female , Male , Rats , Brain , Metabolism , Cholecystokinin , Genetics , Drug Overdose , Food, Formulated , Gene Expression , Hyperphagia , Iodine , Toxicity , RNA, Messenger , Genetics , Rats, Wistar , Thyroid Hormones , Blood , Thyrotropin , Blood , Thyroxine , Blood , Triiodothyronine , BloodABSTRACT
The recent identification of receptor activator of nuclear factor-kappaB ligand (RANKL)/RANK/osteoprotegerin (OPG) cytokine system has led to a new molecular perspective on osteoclast biology and bone homeostasis. Specifically, the interaction between RANKL and RANK is responsible for osteoclast differentiation. In the present study, we evaluated whether soluble RANK (sRANK) could act as an antagonist of RANKL and down-regulate osteoclastogenesis and bone resorption in vitro. The prokaryotic expression vector coding for sRANK was constructed. Then the construct was introduced into E. coli Origami B (DE3) competent cells and recombinant sRANK was successfully produced and purified through affinity chromatography. sRANK reduced osteoclast-like cell (OLC) formation and resorption pit formation induced by parathyroid hormone (PTH) in a dose-dependent manner. In addition, sRANK significantly inhibited PTH-induced mRNA expression of carbonic anhydrase II and tartrate-resistant acid phosphatase in murine bone marrow cells as confirmed by using semi-quantitative RT-PCR. The down-regulation was highly correlated with the effect of sRANK on OLC formation from marrow cells. These data demonstrate the anti-resorptive effects of sRANK in vitro and highlight the potential of sRANK as a novel therapeutic approach to bone disorders characterized by enhanced bone resorption.
Subject(s)
Animals , Humans , Mice , Bone Marrow Cells , Cell Biology , Bone Resorption , Cell Differentiation , Cells, Cultured , Escherichia coli , Genetics , Metabolism , Osteoclasts , Cell Biology , Osteoprotegerin , Physiology , Parathyroid Hormone , Physiology , RANK Ligand , Physiology , Receptor Activator of Nuclear Factor-kappa B , Genetics , Physiology , Recombinant Proteins , Genetics , PharmacologyABSTRACT
Recombinant human BMP-2 was compounded with chitosan/gelatin/hydroxyapatite(HCG) scaffold and the complex was sterilized by 60Co radiating. Osteoblast isolated from cranial bones of newborn rat was primary cultured and seeded onto the complexes. 3 days after culturing, scanning electron microscope(SEM) was applied to detect the compatibility of the cell with the complex. SEM showed osteoblast attached closely with the complex and grew well in its pores. Then the complexes with osteoblast modification were implanted into athymic nude mice subcutaneously. 8 weeks after implantation, X-ray photograph and histological observation were applied to detect the bone formation of the complexes. Under X-ray a high-density areas consistent with the shape of the implanted complex could be seen. Histological observation also proved there was bone formation in the interspace of the complex. A conclusion was drawn that rhBMP-2 compounded HCG scaffold had good osteogenesis ability in vivo.
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<p><b>OBJECTIVE</b>To study the therapeutic effects of Rhizoma Drynariae and estrogen on osteoporosis in ovariectomized (OVX) rats.</p><p><b>METHODS</b>Fifty-five female rats were randomly divided into 5 groups, the normal control (normal),the sham operated (sham), the model, the estrogen, and the Rhizoma drynariae (RD) groups; ovariectomized rats were used as postmenopausal osteoporosis model. The changes of morphology and dynamic parameters in different groups were determined by bone histomorphometry.</p><p><b>RESULTS</b>Compared with the model group, the trabecular volume (TBV/TTV) , trabecular thickness (MTPT) and density (MTPD) in the other four groups were significantly increased, while the trabecular template spacing (MTPS) and the ratio of trabecular surface to trabecular volume (TBS/TBV) significantly decreased (P <0. 05); and the osteoid surface (TOS), single label surface [Sfract (s) ] ,double label surface [Sfract (d) ] and bone formation rate (Svf) also decreased,while osteoid maturation period (OMP) increased in the latter four groups. No significant difference of cortical width (MCW) was found between these 5 groups. Compared with the normal and sham groups, TOS, Sfract ( s) , Sfract ( d) , Svf in the estrogen and RD groups increased significantly, while OMP decreased; no significant difference was found in other parameters.</p><p><b>CONCLUSION</b>Rhizoma Drynariae has the similar effect with estrogen in maintaining normal trabecular structure and connection by inhibiting the increased bone turnover of postmenopausal osteoporosis.</p>
Subject(s)
Animals , Female , Humans , Rats , Drugs, Chinese Herbal , Therapeutic Uses , Estrogens , Therapeutic Uses , Osteoporosis , Drug Therapy , Ovariectomy , Polypodiaceae , PostmenopauseABSTRACT
<p><b>OBJECTIVE</b>To clarify the effects of nylestriol on microarchitecture and interleukin-6 (IL-6) mRNA expression in tibial bone in ovariectomized rats.</p><p><b>METHODS</b>30 female rats were randomly allocated into 3 groups: sham, OVX and nylestriol-treated group. Nylestriol-treated group were ovariectomized, then fed with nylestriol for 3 months and the bone mineral density (BMD) was measured in lumbar vertebra by dual energy x-ray absorptiometry. After sacrifice of the animal, bone histomorphometric parameters were measured to study the changes in bone microarchitecture, and RT-PCR was performed to detect the expression of IL-6 mRNA in bone tissue.</p><p><b>RESULTS</b>BMD was significantly reduced, while IL-6 mRNA level elevated in the OVX group compared with the sham group. Static histomorphometric data showed that the trabecular bone volume, mean trabecular plate thickness and density were reduced while the mean trabecular plate space elevated remarkably in the OVX group in comparison with that in the sham group. As for dynamic parameters, trabecular osteoid surface, tetracyclin labeled surface and bone turnover rate were increased while osteoid maturation rate decreased significantly in the OVX group compared with the sham group. BMD, IL-6 mRNA expression and bone histomorphometric parameters were improved in nylestriol-treated rats.</p><p><b>CONCLUSION</b>Nylestriol plays an important role in maintaining bone volume and improving bone microarchitecture by markedly inhibiting bone turnover and bone resorption, which might be to some degree attributed to reduced IL-6 expression.</p>