Objective To discuss the value of percutaneous dilatational tracheotomy(PDT) for cardiosurgery in intensive care unit by comparing its outcomes with that of open tracheotomy(OT).MethodsA prospective study was conducted on 40 severe patients undergoing cardiosurgery.The patients were divided into PDT(treated from January 2002 to July 2003) and OT groups(treated from August 2003 to February 2008) with 20 cases in each.The operation time,length and heal time of the incision,blood loss,vital signs,and postoperative complications were compared between the two groups.ResultsThe operation time,length and heal time of the incision in the PDT group were significantly different from that in the OT group [(8.8?2.2) min vs(21.5?5.3) min,t=-9.897;(1.3?0.2) cm vs(3.4?0.5) cm,t=-17.440;and(3.9?0.9) d vs(5.9?1.2) d,t=-5.963;P=0.000].One patient in the PDT group(5%) had Ⅱ degree blood loss during the operation that was significantly less than those in the OT group(13 patients,65%,?2=15.824,P=0.000).The level of the vital signs change in the PDT group was significantly milder than those in the OT group [MAP: 1-5 mm Hg(median 2) vs 1-20 mm Hg(median 3.5),Z=-2.959,P=0.003;heart rate: 3-12 beats/min(median 5.5) vs 7-70 beats/min(median 10),Z=-3.956,P=0.000;SpO2: 0-4%(median 2%) vs 0-31%(median 3.5%),Z=-3.548,P=0.000].Furthermore,the rate of incisional leakage of sputum and overall rate of complications in the PDT group were significantly lower than those in the OT group.[1(5%) vs 8(40%),?2=5.161,P=0.023;and 1(5%) vs 12(60%),?2=13.789,P=0.000].ConclusionsPDT is a safe and minimally invasive technique with a high rate of success and low rate of complications.The method is easy to operate,and thus could be a good choice for severe patients after cardiovascular surgeries.

Hematologic Diseases ; Hemolysis ; Humans ; Infant, Newborn

To construct a system of genetic transformation suitable for Rhizopus oryzae, we constructed a single-exchange vector pBS-hygro carrying hygromycin B resistance gene (hph) as its selective marker using gene splicing by overlap extension PCR (SOE PCR) technique. We introduced this recombinant vector into Rhizopus oryzae AS 3.819 by PEG/CaCl2-mediated transformation of protoplast, electroporation of protoplast and germinated spores; and we studied the effects of hydrolysis time, field strength and spore germination time on transformation frequency. We conducted quantitative real-time PCR (qPCR) assay to determine the gene copy number of ldhA integrated in the genome of R. oryzae transformants and its effect on the stability of transformants. We successfully achieved R. oryzae transformants integrated with pBS-hygro-ldhA vector. The optimal hydrolysis time for protoplast production was 140 min, and the optimal field strength of electroporation pulse for protoplast was 13 kV/cm. The optimal germination time of spores for electroporation was 2.5 h, and the optimal field strength of electroporation pulse was 14 kV/cm. The transformation frequency of method based on germinated spores was generally higher than the methods based on protoplast. The qPCR test results suggested that transformants with high copy number of integration in a certain range were relatively stable. Our results provided basis and support for metabolic regulation and genetic engineering breeding of R. oryzae.
DNA, Recombinant ; Electroporation ; Genetic Engineering ; Genetic Vectors ; Hygromycin B ; Protoplasts ; Real-Time Polymerase Chain Reaction ; Rhizopus ; genetics ; Transformation, Genetic

Objective Based on the immunologic balance, to investigate the mechanisms of Danchi decoction for controlling the recurrence of endometriosis (EMs) of qi stagnation and blood stasis after conservative operation. Methods A multi-center and randomized clinical trial was adopted. EMs patients were divided into Chinese medicine group (CG, 20 cases) and western medicine group (WG, 10 cases). The two groups were given Danchi decoction and GnRH-a or gestrinone respectively for 3 months. The serum IL-2, IL-6 and IL-2/IL-6 were detected before operation, after 3 months of treatment and 3 months after withdrawal, and compared with the control group (5 cases of simple ovarian cysts). Results Before operation, the serum IL-2 level of CG and WG was lower than control group (P<0.05), IL-6 was higher than control group (P<0.05), the ratio of IL-2/IL-6 was lower than control group (P<0.05). After treatment and 3 months after withdrawal, IL-2 level of CG and WG was increased than before operation (P<0.05), IL-6 was decreased (P<0.05), IL-2/IL-6 ratio was increased (P<0.05). There was no significant difference in IL-2/IL-6 between CG and control group (P>0.05), but 3 months after withdrawal, IL-2/IL-6 of WG was significantly lower than that of control group (P<0.05). Conclusion Immune imbalance is closely related to the occurrence and development of EMs. Danchi Decoction or GnRH-a or gestrinone can prevent the recurrence of EMs by adjusting the disorder of immune response, and Danchi Decoction has better effect on long-term result.

Objective To evaluate the method of detecting antibodies to Bacillus anthracis by enzymelinked immunosorbent assay(ELISA)using crude antigen.Methods The anti-Bacillus anthracis antibody levels in sera of 42 healthy people and 42 patients were detected by indirect ELISA.Standard curve was plotted using the data from positive controls,based on which the relative content of each serum was calculated and compared with the result of rLF.Results The median of antibody's relative content in patient group and healthy people group are 1.19 and 0.24,the differences being statistically significant(uc=7.643,P＜0.05).The result of crude antigen is in concordance with rLF(but not parallel absolutely).Conclusions Crude antigen can distinguish most of patients with healthy population effectively.The results suggested that crude antigen is applicable in anti-Bacillus anthracis antibody surveillance.

Objective To investigate the levels of IL-6 in serum and gastric mucosa of servicemen before and after mili-tary maneuver and their significance .Methods The level of serum IL-6 in 60 servicemen was studied by enzyme-linked immunosorbent assay .The expression of IL-6 in gastric mucosa from the 60 servicemen was detected by immunochemical staining.Results The level of IL-6 in serum of servicemen after military maneuver was significantly higher than that before military maneuver (P<0.01).IL-6 was found to be expressed in the cytoplasm of epithelioglandular cells and also in the cytoplasm of mesenchymal cells .The expression of IL-6 in gastirc mucosa was significantly stronger after military maneuver than before military maneuver (P<0.01).Conclusion IL-6 might be involved in the development of pathological changes in gastric mucosa caused by military stress in troops .Examination of the level of IL-6 in the early stage of military stress might help assess the chance of gastric mucosa lesion and its severity .

Objective To investigate the expression and distribution of plasmacytoid dendritic cells(pDC) in peripheral blood and renal tissues in children with Henoch-SchSnlein purpura(HSP),and explore the role of pDCs in the pathogenesis of Henoch-Schtnlein purpura nephritis(HSPN).Methods Among the 40 children with HSP,28 cases were in the active phase(renal biopsy performed in 8 cases of them) and the other 12 in remission phase.Peripheral blood mononuclear cells were isolated,and the expression of pDC was detected by flow cytometry.The normal control group was established (n =15).Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression(indicated as 2-△Ct value) of CXC motif chemokine 10 (CXCL10),CC chemokine ligand 5 (CCL5),chemokine CXC subfamily receptor 3 (CXCR3),CC chemokine receptor 5 (CCR5) in children with HSP and those in the controls.Immunohistochemistry labeling technique was used to detect the distribution of pDC in renal tissues from renal biopsy,and the normal controls were established (n =3).Results The expression percentage of pDC in peripheral blood in active phase was 0.051 ± 0.039,significantly lower than those in remission phase (0.181 ± 0.082) and the normal controls (0.166 ± 0.079) (P ＜ 0.000 1).Chemokines genes CXCL10 and CCL5 were overexpressed in peripheral blood ceils of acute phase HSP children,but chemokine receptors CXCR3,CCR5 were lowly expressed compared with normal controls.There was almost no expression of pDC in the normal control renal tissues,while pDC was infiltrated in glomeruli of HSPN children.Conclusions The number of pDC and chemokines' expression in peripheral blood is abnormal,and the pathogenesis of nephritis may be involved with the pDC in peripheral blood to migrate to the renal tissues.

OBJECTIVETo observe the effect of detoxifying and blood circulation activating Chinese herb extraction of polygonum cuspidatum and hawthorn on carotid intima-media thickness (IMT), plaque integral and plaque stability related serum indexes of patients with carotid atherosclerosis.

METHODSixty and four cases of carotid artery atherosclerosis patients were assigned randomly to 2 groups: detoxifying and blood circulation activating treatment group (treatment group, 32 cases) and control group (32 cases). Patients in treatment group were treated with capsules of extraction of polygonum cuspidatum and hawthorn, 1 pill po, bid (dosage of administration: polygonum cuspidatum extraction 5.33 mg x kg(-1) x d(-1), hawthorn extraction 5.0 mg x kg(-1) x d(-1)); patients in control group were treated with lovastatin 20 mg po, qd (dosage of administration: 0.33 mg x kg(-1) x d(-1)). The course of treatment was six months. To observe changes of IMT, plaque integral, and detect the level of plaque stability related serum indexes such as Hs-CRP, MMP-1 and TIMP-1.

RESULTAfter 6 months of treatment, in control group one patient quit the clinical trial because of liver dysfunction and one patient was rejected because of having not followed the therapeutic regimen. 32 cases in treatment group and 30 cases in control group were analyzed. The results showed that IMT and plaque integral of treatment group decreased significantly after the treatment (P < 0.05, P < 0.01), and there was no significant difference compared with control grope. Serum Hs-CRP, MMP-1 and MMP-1/TIMP-1 decreased after the treatment (P < 0.05, P < 0.01), and the treatment group was superior to control group in decreasing serum Hs-CRP (P < 0.05).

CONCLUSIONDetoxifying and blood circulation activating Chinese herb extraction of polygonum cuspidatum and hawthorn has good effect of anti-atherosclerosis and promoting plaque stability. Its mechanism might be related with anti-inflammation and inhibiting degradation of extracellular matrix, and deserves further studies.

C-Reactive Protein ; metabolism ; Carotid Artery Diseases ; blood ; drug therapy ; Crataegus ; chemistry ; Drugs, Chinese Herbal ; adverse effects ; pharmacology ; therapeutic use ; Fallopia japonica ; chemistry ; Female ; Humans ; Male ; Matrix Metalloproteinase 1 ; blood ; Middle Aged ; Safety ; Tissue Inhibitor of Metalloproteinase-1 ; blood

A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases ; biosynthesis ; genetics ; Azulenes ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Open Reading Frames ; Polyisoprenyl Phosphates ; Recombinant Proteins ; biosynthesis ; Sesquiterpenes ; metabolism ; Sesquiterpenes, Guaiane ; Thymelaeaceae ; enzymology ; genetics

Objective To study the effects of semiconductor laser irradiation and high voltage static electric fields on small vessel damage in diabetic rats.Methods Fifty healthy male Wistar rats were randomly divided into two groups:8 rats in a normal group and 42 in a diabetic model group.The diabetic models were created by intrape- ritoneal injection of streptozocin.The diabetic model rats were randomly divided into four subgroups:a diabetes group,a semiconductor laser treatment group,a high voltage static electric field treatment group and a comprehensive treatment group receiving combined semiconductor laser and high voltage static electric field exposure.The rats in each treatment group were subjected to the corresponding intervention.After 20 days of treatment,the venous blood, kidney tissue and myocardium tissue of the rats were collected,and the concentrations of blood glucose,insulin,en- dothelin and tissue were detected.Results Compared with the normal group,a significant increase in blood glu- cose and endothelin was observed in the diabetic model group,along with significantly decreased blood insulin and significant small vessel endothelium proliferation in the kidney tissue.Compared with the diabetes group,endothelin levels were significantly lower in all 3 treatment groups,and blood insulin was also higher in the comprehensive treat- ment group.Conclusion There were severe abnormalities in blood glucose,insulin and endothelin as well as mild impairment of small vessel endothelium proliferation in the diabetic rats.Semiconductor laser and high voltage static electric field exposure have a role in treating and preventing these conditions in diabetic rats.