OBJECTIVETo investigate the changes of human leukocyte antigen DR (HLA-DR) expression in the monocytes of elderly patients with sepsis.

METHODSA total of 213 elderly patients were divided into 4 groups according to the criteria defined by the American College of Chest Physicians/ Society of Critical Care Medicine (ACCP/SCCM), namely the non-sepsis group (group A), sepsis group (group B), severe sepsis group (group C) and septic shock group (group D). Another 60 healthy subjects were recruited as the control group. HLA-DR expression in the monocytes from each subject was determined by flow cytometry, and the HLA-DR levels were compared between the groups.

RESULTSCompared with that of the healthy control, HLA-DR expression was significantly increased in group A and lowered in group D. No significant differences were found between the control group and groups B and C. HLA-DR expression was significantly different between groups A, B, C, and D (P<0.01), increasing in the order of group D (32.74-/+13.4)%, group C (61.9-/+14.29)%, group B (69.99-/+12.8)%, and group A (85.06-/+15.37)%.

CONCLUSIONSAlthough with multiple organ diseases and repeated or long-term hospitalization, the non-septic elderly patients do not show lowered expression of HLA-DR. In the elderly patients with sepsis and severe sepsis, HLA-DR expression is not a sensitive index, but obviously lowered HLA-DR expression may serve as a specific indicator in elderly patients with septic shock.

Aged ; Aged, 80 and over ; Biomarkers ; analysis ; Female ; Flow Cytometry ; HLA-DR Antigens ; analysis ; biosynthesis ; Humans ; Leukocytes, Mononuclear ; immunology ; Male ; Sepsis ; immunology ; Shock, Septic ; immunology

This study was aimed to investigate the conditions of culturing in vitro mesenchymal stem cells (MSCs) derived from bone marrow of children with acute leukemia and the biological characteristics of MSCs from leukemia children. The bone marrow MSCs of acute leukemia children were isolated by density gradient centrifugation combined with adherent segregating method and cultured in DMEM/F12. The morphology of Wright stained MSCs was observed under inverted microscope. Cell surface markers were analyzed with flow cytometry. The growth characteristic features of cultured MSCs was measured with MTT method. Induced adipogenic and osteogenic differentiation of MSCs in appropriate induction media was observed. The results indicated that BM-MSCs of acute leukemia children could be successfully cultured in vitro in appropriate conditions. At 24 hours of culture the MSCs began to adhere to wall, grew in colony and appeared in different shapes. As the culture lasted, the MSCs proliferated continuously and shaped in fusiform. After 2 - 3 weeks of culture, MSCs covered the bottom of culture flask. The analysis of growth feature showed that MSCs were in latency for 3 days, and then entered into growth period. After 8 days of culture the growth of MSCs showed to be in plateau stage. The shape of MSCs in 1st and 2nd generation showed to be heterogeneous but the 3rd generation to be homogeneous with long-fusiform. Cells were arranged in shape of whirlpool or radiation. The surface marker analysis showed that the MSCs were positive for CD105, CD29, CD13, but negative for CD34, CD45, CD14 and HLA-DR. The MSCs from leukemia children could be induced into adipocytes and osteocytes in appropriate conditions. It is concluded that (1) MSCs derived from children with acute leukemia can be successfully cultured and passaged in vitro; (2) MSCs from leukemia children not received chemotherapy are more successfully cultured in vitro than those received chemotherapy; (3) the common biological characteristics of MSCs from children with acute leukemia are same as the MSCs from healthy person.
Adolescent ; Bone Marrow ; pathology ; Bone Marrow Cells ; cytology ; metabolism ; Cell Culture Techniques ; Child ; Child, Preschool ; Female ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured ; Young Adult

ObjectiveTo introduction of perforator flaps,muscle flaps pedicled by descending branch of lateral circumflex femoral artery,method and their clinical application that multiple soft tissue defects of hand are repaire by muliplefoliated tissue flap only branch lateral circumflex femoral artery.MethodsFifteen patients with multiple soft tissue defects of hand were repaired muliplefoliated tissue flap only pedicled bydescending branch lateral circumflex femoral artery.At first,the anterolateral thigh perforator flap was designed and harvested according to the soft tissue defects of hand, then the descending branch lateral circumflex femoral artery was dissected at the same time the segmented perforator flap,fascia lata flap,rectus femoris muscle flap, vastus lateralis muscle flap, vastus intermedius muscle flap and distal spatium intermusculare flap were harvested in need according to distance among soft tissue defects.The muliplefoliated tissue flap was harvested only pedicled by descending branch lateral circumflex femoral artery, at last muscle flaps and fascia lata flaps were covered by skin graft, so the multiple soft tissue defects of hand were repaired in one time.ResultsNo vascular crisis happened. All skin grafts survived well, the contour of all repaired soft tissue defects was good and protective feeling was recovered by skin grafts of all flaps. All cases were got follow-up and the range was from 6 to 20 months(the average was 8.7 months).Wound of donor site healed well, muscle strength of quadriceps and motion of knee were normal. Three cases were excellent,nine cases were well and 3 cases were good, according to upper extremity function evaluation criteria of Chinese Medical Society for the Surgery of the Hand, the rate of good was 80 percent.ConclusionMultiple soft tissue defects of hand can be repaired by muliplefoliated flap only pedicled by descending branch of lateral circumflex femoral artery. Its advantages included reduction of operation time and treatment, good recovery of hand contour and function. It is a good method to repair multiple soft tissue defects of hand.

Objective:To observe the effects of electroacupuncture (EA) of three different frequencies (2 Hz,80 Hz and 2 Hz/80 Hz) on the free radicals in hippocampus of vascular dementia (VD) model mice.Methods:A total of 100 Kunming mice were randomly divided into a sham operation group,a model group,a 2 Hz EA group,an 80 Hz EA group and a 2 Hz/80 Hz EA group,with 20 mice in each group.The ischemia-reperfusion VD model was established by repeated blockade of bilateral common carotid arteries.Mice in EA groups began EA treatment on the 4th day after the operation.Baihui (GV 20),Dazhui (GV 14),Geshu (BL 17) and Zusanli (ST 36) were punctured and then connected to EA instrument,with different waves of 2 Hz,80 Hz or 2 Hz/80 Hz (10 min/time) applied accordingly,once a day.During the jumping stand experiment,the learning performance,memory performance and hippocampal calcitonin gene-related peptide (CGRP),nitric oxide synthase (NOS),malondialdehyde (MDA),changes in superoxide dismutase (SOD) and true choline esterase (TChE) were observed.In hippocampus,the CGRP level was determined by radioimmunoassay;the MDA level was determined by thiobarbituric acid colorimetric method;the activities of NOS and TChE were determined by spectrophotometry;the activity of SOD was determined by xanthine oxidase method.Results:Compared with the sham operation group,the performances of learning and memory decreased significantly in the model group (P＜0.01);in hippocampus,the CGRP level decreased,the MDA level increased,the activities of NOS and TChE increased,and the activity of SOD decreased in the model group.Compared with the model group,the learning and memory performances of the EA groups were significantly improved (P＜0.05 or P＜0.01);in hippocampus,the CGRP level increased,the MDA level decreased,the NOS and TChE activities decreased,and the SOD activity increased (P＜0.05 or P＜0.01).Among EA groups,the 2 Hz/80 Hz EA group was superior to the 2 Hz EA group and the 80 Hz EA group (P＜0.05 or P＜0.01).Conclusion:EA can improve the cognitive impairment of mice with ischemia-reperfusion VD.The mechanism may be related to the improvement of cerebral blood circulation,regulation of the central neurotransmitters,fighting lipid peroxidation and promoting nerve cell repair.The therapeutic effects of EA with different frequencies were different,and the intervention effect by EA at 2 Hz/80Hz is the most significant.

BACKGROUND: Previous findings show that osteoblast-specific peptides can promote the repair and regeneration of skull defects in rabbits, and β-tricalcium phosphate (β-TCP) is used as a scaffold to carry osteoblast-specific polypeptides. Both of them not only complement each other, but also fully exert dual effects of osteoinduction and bone conduction. OBJECTIVE: To investigate the effect of osteoblast-specific peptide on the preservation of the anterior tooth extraction site in rabbits, and to study the effect on the alveolar bone remodeling. METHODS: Twenty-seven New Zealand white rabbits were randomly divided into three groups (n=9 per group), and the right mandibular incisors were removed to establish the animal models of tooth extraction. β-TCP/osteoblast-specific peptide compounds were implanted in the experimental group, and pure β-TCP meal was implanted into the material group. The blank control group had no implantation. Three rabbits from each group were scarified at 4, 8 and 12 postoperative weeks, and tissue samples were prepared for gross observation, histomorphology measurements, and radiographic measurements of extraction socket healing. RESULTS AND CONCLUSION: (1) Imaging results showed that the relative length of residual alveolar bone after modeling was ranked as follows: the experimental group > the material group > the blank control group, and the difference was statistically significant among groups (P < 0.05). Cone-beam CT findings in the three groups changed as time went on. At 4 and 8 postoperative weeks, the implanted materials in the experimental and material groups gradually degraded; the bone mass in the experimental group was significantly higher than that in the material and blank control groups. At 12 postoperative weeks, the experimental group had basically completed the reconstruction of tooth socket, but there were still some bone defects in the material and blank control groups. (2) Histomorphological findings showed that at 4 postoperative weeks, the experimental group exhibited obvious bone deposition lines, and the bone trabecula was widened; in the material and blank control groups, the new bone was less. At 8 postoperative weeks, a small amount of undegraded scaffold was found in the experimental group, with mature lamellar bone, the amount of new bone tissues in the material group increased and osteoblasts were obviously detected in the blank control group. At 12 postoperative weeks, the bone remodeling in the extraction socket of the experimental group was basically completed; in the material group, there were still a small amount of scaffold materials and dense plate-like new bone; and in the blank control group, the new bone tended to be mature, and there was obvious lamellar structure. To conclude, osteoblast-specific peptides can effectively preserve the length of the residual alveolar bone after tooth extraction, promote the formation of new bone, and have the function of preserving the tooth extraction site.

OBJECTIVETo investigate the factors related to the outcome of patients with chronic severe hepatitis B and effectiveness of antivirus therapy.

METHODSThe effects of the factors including age, prothrombin activity (PTA), serum HBeAg, Anti-HBe, HBV-DNA load, with or without complication, antivirus therapy and so on, on outcome of 330 patients with chronic severe hepatitis B were analyzed in this retrospective study.

RESULTSThe mortality of patients with chronic severe hepatitis B was significantly higher among patients at higher age, with lower PTA, and with more complications. The mortality of patients with HBV-DNA more than 1x10(5) copies/ml (52.3 percent) was higher than that of patients whose HBV-DNA was less than 1x10(5) copies/ml (32.9 percent). There was no correlation between serum HBeAg or anti-HBe and the mortality. The mortality of patients with HBV-DNA higher than 1x10(5) copies/ml (30.38 percent) who were treated with lamivudine in 2005 was lower than that of patients whose HBV-DNA was less than 1x10(5) copies/ml (54.64 percent) who were not treated with any antiviral therapy in 2001.

CONCLUSIONThe higher serum virus load is the key factors of the mortality in addition to the other factors such as older age, lower PTA, more complication in the patients with chronic severe hepatitis B. The usage of antivirus therapy may be associated with lower mortality.

Adult ; Age Factors ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; metabolism ; mortality ; Humans ; Lamivudine ; therapeutic use ; Male ; Middle Aged ; Prothrombin ; metabolism ; Treatment Outcome ; Viral Load

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

The paper is to report the study of pharmacokinetics of transdermal administered nicotine in the brain of freely moving rat by using microdialysis with stable labeled isotope as internal standard. The pharmacokinetic behavior of nicotine in Sprague Dawley rat brain was investigated after intranasal administration (3.75 mg). Brain fluid samples were collected by intracerebral microdialysis with DL-nicotine as internal standard. Concentrations of nicotine and DL-nicotine in the sample were measured by HPLC-MS/MS. Main pharmacokinetic parameters were calculated and analyzed by Das 2.0 pharmacokinetic software. The recovery of nicotine and the delivery of DL-nicotine were the same. The fate of absorption and distribution was two compartment model and the values of t1/2alpha was 170.31 min, t1/2beta was 263.30 min and the AUC(0-infinity) was 2.75 x 10(5) microg x L(-1) min separately. DL-nicotine can be used to calibrate the recovery of nicotine, and the new method of stable isotope microdialysis can be used to study the pharmacokinetics of freely moving rat. It will make sense for the treatment of addiction of tobacco and provide a new thought for the research of pharmacokinetics-pharmacodynamic combination.
Administration, Cutaneous ; Administration, Intranasal ; Animals ; Area Under Curve ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Deuterium ; Female ; Isotope Labeling ; methods ; Male ; Microdialysis ; methods ; Nicotine ; administration & dosage ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

Objective To investigate the influence of early growth response gene-1 (EGR1) on the autophagy of host cells following infection with human T cell leukemia virus type 1 (HTLV-1).MethodsA HTLV-1-positive cell line MT2 was co-cultured with HeLa cells for 24 h to construct the virus early infection model.Immunoblotting assay was used to detect the expression of HTLV-1 core protein p19 and EGR1.Luciferase reporter gene analysis was used to detect the transcriptional activity of 5′-regulatory sequence of EGR1 at different time points after co-culturing.An effective small interfering RNA (siRNA) targeting EGR1 was screened out and transfected into HeLa cells by Lipofectamine 2000.Then the transfected HeLa cells were co-cultured with the HTLV-1-positive cell line MT2 for 24 h.Immunoblotting assay was used to detect HTLV-1 core protein p19, EGR1 and autophagy-related protein LC3.Real-time PCR was performed to detect viral load.Autophagosome was analyzed by immunofluorescence after co-culturing.Results The expression of EGR1 and the transcriptional activity of pEGR1-luc gradually increased after co-culturing HeLa cells with MT2 cells for 8 h (P<0.01).The expression of EGR1 was positively correlated with host cell autophagy following HTLV-1 infection.The effective siRNA for silencing the expression of EGR1 was obtained and named as siE2.The viral load, the expression of HTLV-1 core protein p19 and the proportion of LC3B/LC3A in the co-culture model were markedly down-regulated by RNA interference with siE2, which was concomitant with a persistent decrease of intracellular autophagosome (P<0.01).Conclusion EGR1 is associated with host cell autophagy and viral replication in HTLV-1 infection.