OBJECTIVETo investigate the technological parameters of the purification process of total flavones from Smilax china with macroporous absorption resin.

METHODThe technical process for purification of total flavones with the optimum macroporous absorption resin was screened by yield of total flavones product.

RESULTThe D140 macroporous absorption resin had the best separating efficiency when the flavones content in the liquid was 0.5 g x mL(-1) equivalent to raw material, the volume of drug 18 BV (resin bad volume) with the adsorption-power 2 BV x h(-1), and the volume of 60% (mL x mL(-1)) ethanol as eluant 5-10 BV (resin bad volume) with desorption-power 1 BV x h(-1). The obtained flavones product has total flavones recovery rate of 84.72%.

CONCLUSIONThe treatment of regenerated resin is easy, this method is advisable.

Absorption ; Flavones ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Resins, Synthetic ; chemistry ; Rhizome ; chemistry ; Smilax ; chemistry ; Technology, Pharmaceutical ; methods

Objective To explore the ability for constructing tissue engineering bone in vivo in complex scaffolds with PHB‐HHx as the scaffolds material and human umbilical cord mensenchymal stem cells (hUCMCs) as the seed cells .Methods hUCM‐SCs were inoculated into PHBHHx scaffolds to induce osteogenesis culture in vivo for two weeks ,the the induced group was the experimental group and those without instilling hUCMCs served as the control group ,the nude mouse subcutaneous implantation was performed .Then taking material at 1 ,3 ,5 months after implantation in vivo was performed for conducting HE ,collagenⅠim‐munohistochemical ,alkaline phosphatase staining and RT‐PCR .Results hUCMSCs showed good cellular adsorbability .The size and form in the experimental group basically maintained the original status ,and the osteogenesis specific indicators were positive ;but the control group did not keek the original status ,its volume was gradually shrunk until complete degradation ,and the osteogen‐esis specific indicators were negative .Conclusion The PHBHHx scaffolds combined with hUCMSCs has the capability of in vivo heterotopic constructing tissue engineering bone in nude mouse by in vitro osteogenic induction .

Objective To study the effect of autoantibody test which includes antinuclear antibodies(ANA),antinuclear antibody fluorescence patterns,anti-extractable nuclear antigen(ENA) antibodies and anti-double strands DNA(ds-DNA) antibodies in the diagnosis of pediactic autoimmune diseases.Methods Of all the inpatients which had positive results of autoantibody tests,135 cases were reviewed.The autoantibody assay,positive value(PV) analysis were performed respectively.Results PV of ANA test to autoimmune diseases was 0.36 which was proportional to the intensity of fluorescence;Of all the fluorescence patterns,speckled(fine) had a relatively high PV;Anti-ENA and anti-dsDNA antibody tests had higher PV than ANA test.Conclusion Fluorescence intensity,(anti-ENA) antibody test and anti-dsDNA antibody test may be useful in identifying autoimmune diseases in clinic.

OBJECTIVETo study the effect of bifidobacterium genomic DNA on umbilical cord blood mononuclear cell (CBMC), and investigate the immunoregulation of bifidobacterium DNA and explore possible mechanisms by which bifidobacterium acts against allergic reaction.

METHODSBifidobacterium genomic DNA (bDNA) and human DNA (hDNA) were extracted with phenol/chloroform/isoamyl alcohol and stored at -20 degrees C for later use. Parts of bDNA were completely digested with DNaseI (d-bDNA) at 37 degrees C. CBMCs were isolated with Ficoll from umbilical cord blood and incubated at 37 degrees C in a 5% CO2 humidified incubator. These cells were divided into four groups, control group: without any stimulant; bDNA group: stimulated with 25 microg/ml bDNA; d-bDNA group: stimulated with 25 microg/ml d-bDNA; hDNA group: stimulated with 25 microg/ml hDNA. The cells were stimulated with different stimulants in vitro, at the end of incubation culture supernatant and cells were collected. IL-12 and IL-10 levels in the culture supernatant were measured by enzyme linked immuno sorbent assay (ELISA); cells secreting IL-4 and IFN-gamma were counted by enzyme linked immunospot (ELISPOT) assay; and total RNA was isolated from the cells to assay T-bet and GATA3 mRNA expression levels by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSSix hours after stimulation there was no significant difference in IL-12 level in supernatant among the four groups; 12 hours after stimulation, IL-12 level in supernatant of bDNA treated group was significantly higher than that of each of the other groups, so were the results obtained at 24 hours and 48 hours after stimulation (P < 0.05). No significant difference could be detected in IL-12 level in supernatant among the other 3 groups. On the other hand, 6 hours after stimulation there was no significant difference in IL-10 level in supernatant among the four groups. But 12 and 24 hours after stimulation IL-10 level in supernatant of bDNA treated group was lower than that of each of the other groups, but the difference was not statistically significant. The count of IFN-gamma secreting cells of bDNA treated group was higher than that of the other groups, while IL-4 secteting cells of bDNA treated group were lower than that of the other groups. After bDNA stimulation, nuclear factor T-box expressed in T cells (T-bet) mRNA expression was conspicuously enhanced as compared to the other three groups (P < 0.05). GATA3 mRNA transcription in CBMC had no significant change after bDNA stimulation.

CONCLUSIONbDNA could promote secretion of Th1 type cytokine IL-12, while Th2 type cytokine IL-10 level of cell supernatant was decreased. bDNA could stimulate secretion of IFN-gamma by CBMC and inhibit secretion of IL-4. T-bet mRNA expression was highly enhanced after bDNA stimulation. bDNA could upregulate Th1 type response, which may be one of important mechanisms by which bifidobacterium inhibit allergic response.

Bifidobacterium ; cytology ; genetics ; Cell Culture Techniques ; DNA, Bacterial ; biosynthesis ; metabolism ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; immunology ; GATA3 Transcription Factor ; genetics ; Humans ; Infant, Newborn ; Interferon-gamma ; immunology ; secretion ; Interleukin-10 ; immunology ; secretion ; Interleukin-12 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Leukocytes, Mononuclear ; immunology ; secretion ; RNA, Messenger ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; T-Box Domain Proteins ; genetics ; Th1 Cells ; drug effects ; immunology ; secretion

OBJECTIVE: To explore alcohol pharmacokinetics as well as acetaldehyde level in peripheral blood in human subjects with different ALDH2 genotypes after drinking. METHODS: Venous blood samples of 14 unrelated volunteers were collected. Polymerase chain reaction-restriction fragment length polymorphism technology was adopted for DNA extraction and ALDH2 genotyping. The volunteers were asked to drink beer at certain doses. The concentration of alcohol and acetaldehyde were assayed by headspace gas chromatography method at different time. The pharmacokinetic parameters were calculated. RESULTS: According to the results of electrophoresis, 5 people carried ALDH2*1/*1 as wild group and 9 people carried ALDH2*1/*2 as mutation group. The good linear range of alcohol and acetaldehyde were 0-1 570.7 microg/mL and 0-5.1772 microg/mL, respectively. The AUC values of alcohol and acetaldehyde and the t1/2Z value of alcohol were higher in the mutation group than that in the wild group. But the CL/F value of alcohol was lower in the mutation group than that in the wild group (P<0.05). CONCLUSION After the consumption of alcohol, alcohol and acetaldehyde metabolism in blood slow down in ALDH2*1/*2 mutation group influenced by the inhibition of enzyme activity, leading to the accumulation of acetaldehyde in peripheral blood, thus reinforcing their effects in the body.
Alcohol Drinking ; Aldehyde Dehydrogenase/genetics* ; Aldehyde Dehydrogenase, Mitochondrial ; Ethanol/metabolism* ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic

Objective To summarize the experience of long-term survival in patients after simulta- neous kidney-pancreas transplantation(SKPT)with modified enteric drainage(ED).Methods From October 2001 to July 2004,6 patients with end-stage renal disease due to Type 1 diabetes underwent SKPT with modified ED,ie,side-to-side anastomosis between the duodenum of donors and jejunum of recipients. The medication regimen included:mycophenolic acid 500 mg and tacrolimus 2 mg before operation;methyl- prednisolone(MP)1.0 during operation;and 2-dose anti-IL-2 receptor monoclonal antibody(2 cases)or antihuman thymocyte globulin(ATG)(4 cases)for immune induction therapy;MP was used on the first 3 d after transplantation,triple immunosuppressive therapy(tacrotimus,mycophenolic acid and prednisone)was used on the second d after transplantation.Anticoagulants such as low molecular heparin or alprostadil were used for 7-10 d to prevent thrombosis in pancreas graft.Somatostatin was used as prophylaxis for graft pan- creatitis.Ganciclovir was used to prevent cytomegalovirus infection when renal graft gradually recovered 3 to 5 d after transplantation.The follow-up was from 1 year and 3 months to 4 years and 1 month.Results Transplantation was successful in all 6 cases.The blood sugar levels were 6-16 mmol/L.Low-dose insulin was used for 5-10 d,then the blood sugar levels returned to normal range.One of 6 patients experienced nephrotoxicity because of high tacrolimus blood concentration at 7 d after operation;after 3 dialyses and re- duction of tacrolimus dose,the renal allograft regained normal function.Three cases experienced alimentary tract hemorrhage at 14,20 and 22 d,respectively,after operation;the bleeding was stopped after treatment. There were no complications such as pancreatic fistula,intestinal fistula and thrombosis early after operation. All the patients are now alive,specifically,1 survived over 4 years,3 over 3 years,1 over 2 years,and 1 over 1 year.All had normal blood sugar free of insulin use.Five cases had normal renal graft function,with normal sCr,and 1 had sCr＞400?mol/L. Two cases were admitted to hospital due to upper respiratory infection and furuncles in the skin of head 6 months and 2 years,respectively,after operation.They were both cured.No complications such as urinary infection,metabolic acidosis and dehydration occurred.Conclusions SKPT is effective for the treatment of end-stage renal disease due to Type 1 diabetes.SKPT with modified ED are relatively simple with physiological compatibility and fewer complications.High quality of donated organs, HLA matching,pancreatic drainage pattern,rational periopcrative medications and infection late after trans- plantation are important factors affecting the long-term survival of the patients.

OBJECTIVETo study the morphological, ultramicrostructural and pathological changes of tissues from a patient with severe acute respiratory syndrome (SARS).

METHODSOne autopsy case of diagnosed SARS was investigated. Lung puncture was performed immediately after the patient died, and the autopsy was done after 12 h. The specimens from lymph nodes, spleen, small intestine, colon and bone marrow were studied by immunohistochemical technique. The antibodies used included CD20, CD45RO (UCHL-1), CD4, CD8, CD68 and CD34.

RESULTSThe principal lesions of the SARS case consisted of acute lobular intrastitial pneumonia, hyaloid membranes of pulmonic alveoli and hyperplasia and shedding of alveolar epithelium of. Virus-like inclusions occasionally contained cytoplasm of the alveolar epithelium, which were positive by histochemical staining. The adjacent blood-vessels were changed by hyperplasia and enlargement. The structures of lymph nodes and spleen were damaged with lymph follicles depletion and splenic nodules atrophy. The specific changes included reduction of lymphocytes and hyperplasia of histiocytes, depletion of the follicles of small intestine and colon wall, decreased hyperplasia of the bone marrow and increased number of the megakaryocyte. Meanwhile, in the immunohistochemical study, CD(20)(+) B cells were fully expressed in lymph nodes and spleen, and the CD45RO (UCHL-1)(+) T cells were scatteredly expressed. The number of CD4(+) help T cell was markedly decreased, while the number of CD8+ poisonal T cells increased, and the ratio of the former and latter was no more than 0.5. Under the electronic microscopy observation, virus-like particles with 80 - 160 nm diameter and halo or garland envelope were found in mononuclear macrophage and cytoplasm of alveolar epithelium.

CONCLUSIONThe specific lesions of SARS consist of lobular intrastitial pneumonia with the formation of hyaline membranes of lung, haemorrhage, necrosis, inflammation of blood vessels and the damages of extralung lymphohemopioetic system. The damages were very similar to the pathological features of tissues infected by human immunodeficiency virus, in which numbers of T cells decreased and CD(4)(+) T cell/CD(8)(+) T cell ratio was no more than 0.5. According to the virus-like particles found in lung of the SARS case, it is considered that these virus-like particles may be a new kind of coronavirus which caused the "atypical pneumonia".

Humans ; Immunohistochemistry ; Lung ; pathology ; Lymph Nodes ; pathology ; Male ; Microscopy, Electron ; Middle Aged ; Myocardium ; pathology ; Severe Acute Respiratory Syndrome ; pathology

OBJECTIVETo study the influence of separation of distal tibiofibular syndesmosis on ankle joint and to compare various operative methods so as to find suitable stabilization for separated distal tibiofibular syndesmosis.

METHODSFrom July 1997 to July 2002, we treated 87 patients (64 males and 23 females, aged 18-54 years) with separation of distal tibiofibular syndesmosis, among whom, 79 were combined with fracture of malleolus. Manipulative reduction, internal fixation with cancellous screws and external fixation with plaster support were performed on 37 patients, fixation with plate and screws for fibular fracture and fixation with cancellous screws for distal tibiofibular syndesmosis on 34 patients, and repair of the distal tibiofibular ligaments with tendon of peroneus longus, reduction of the separated distal tibiofibular syndesmosis, and fixation with cancellous screws on 16 patients. The ankle joint had been dorsiflexed for 30 degrees when the distal tibiofibular syndesmosis was fixed with cancellous screws. And the cancellous screws were taken out at 8-10 weeks after operation.

RESULTSThese patients were followed up for at least two years. The curative effects were assessed according to the complaints of the patients and the contour, function and radiogram of the ankle joint: excellent in 55 patients (63%), good in 18 patients (21%), and fair in 14 patients (16%). Separation of distal tibiofibular syndesmosis recurred in 2 patients, who underwent a reoperation for repairing the distal tibiofibular ligaments with tendon of peroneus longus and recovered. One cancellous screw was broken off. No necrosis developed in the anterior skin of the ankle mortise.

CONCLUSIONSSeparation of distal tibiofibular syndesmosis can be treated with various reasonable operations. Repair with tendon of the peroneus longus can get excellent outcomes for complete separation of the distal tibiofibular syndesmosis.

Adolescent ; Adult ; Ankle Injuries ; diagnostic imaging ; physiopathology ; surgery ; Bone Screws ; Casts, Surgical ; Female ; Fibula ; injuries ; physiopathology ; Follow-Up Studies ; Humans ; Ligaments, Articular ; injuries ; physiopathology ; Male ; Middle Aged ; Radiography ; Tibia ; injuries ; physiopathology ; Treatment Outcome

This study was to determine the protective effect of ω-3 polyunsaturated fatty acids (ω-3PUFAs) on MK-801-induced cognitive impairment in schizophrenia (SZ) rats and the underlying mechanism.A rat model of schizophrenia was induced by MK-801.The cognitive function of rats was assessed using a Morris water maze.The number of hippocampal neurons was measured by Nissl staining.The expression of CREB,p-CREB,BDNF,TrkB,p-TrkB,AKT,p-AKT,ERK,and p-ERK in the hippocampus of rats was detected by Western blotting.The results showed that ω-3PUFAs attenuated MK-801-induced cognitive,impairment and hippocampal neurons loss,reversed the injury of the CREB/BDNF/TrtB pathway induced by MK-801,and antagonized MK-801-induced down-regulation of p-AKT and p-ERK in the hippocampus of rats.In conclusion,ω-3PUFAs enhances the CREB/BDNF/TrkB pathway by activating ERK and AKT,thereby increasing the synaptic plasticity and decreashng neuron loss,and antagonizing MK-801-induced cognitive impairment in schizophrenic rats.

Objective To explore the relationship between post-traumatic stress disorder (PTSD) and perceived social support (PSS)ability among middle school students in earthquake-stricken areas after the Wenchuan earthquake accident. Methods A total of 1966 students from three secondary schools of Wenchuan earthquake-stricken areas were evaluated by PTSD Checklist-Civilian Version (PCL-C) and Perceived Social Support Scale (PSSS). Results The overall incidence rate of PTSD was 78.3%, with severe PTSD as 24.38%. Significant differences on the incidence rates of PTSD were found among the students who were in different PSS levels (P<0.05) and significant negative correlation existed between the levels of severity on PTSD and PSS (γ=-0.226, P<0.05). Significant differences on PTSD incidence rates were found among those students who were from different families or out-family PSS levels (P=0.009, P<0.05). Significant negative correlation existed between the severity of PTSD and family or out-family PSS level (γ=-0.176, P<0.05, γ=-0.214, P<0.05). Conclusion Relationships between the incidence rate, severity of PTSD and PSS levels existed among the middle school students in Wenchuan earthquake-stricken areas, with higher PSS, lower incidence rate and lighter severity of PTSD. Psychological intervention for earthquake-stricken students should be carried out.