Objective To investigate the effect of hypertonic sodium chloride hydroxyethyl 40 injection (HSH)on craniocerebral injury accompanied with hypovolemic shock and ICP.Method Sixty patients suffering from craniecerebral injury accompanied with the hypovolemic shock and admitted to ICU and Neurosurgical Department of Guang dong General Hospital from September 2007 to June 2008,were chosen into the study prospectively.Those who was younger than 18years or older than 70 years,and those who were pregnant or meustruous women,or serious hepatic and renal insufficiency,hypertension,coronaxy artery disease,diabets mellitue,uncontrolled bleeding,brain death,erc were excluded.The patients were randomly divided into 3 groups.Each group included 20 patients and they were resuscitated with LR(500 ml),HIS(4 ml/kg)and HSH(4 ml/kg)respectively.The changos of blood pressure,laboratory examination indices and ICP Were detected before and at 30,60,120 minutes after resuscitation.Results MAP raised over 60 mmHg and ICP declined 10%within 30 min in HIS and HSH groups.The effect of HSH on the improving of blood pressure and the reduction of ICP wag more obvious than that of LR and HIS at 120 minutes(F=18.43,8.99,P<0.05)after resuscitation.There were no obvious oboes of the laboratory examination indices after resuscitation.Conclusions The themputic effect of HSH on ICP and MAP in craniocerebral injury accompanied with hypovolemic shock is obvious and lasting,which would be beneficial to the protection of cerebral function.

Objective To observe different effects of cyclic and continuous nutrition infusion on serum nutritional indicators, peripheral white blood cell counts (WBC), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2) and acute physiology and chronic health evaluation Ⅲ (APACHE Ⅲ ) score in patients with mechanical ventilated respiratory failure (MVRF).Methods A cross-over and self-controlled trial was conducted in 48 patients with MVRF treated in a medical intensive care unit during December 2006 to June 2009, and continuous nutrition (group A) and cyclic nutrition (group B) were infused respectively for patients of the two groups.Serum levels of albumin (ALB), pre-albumin (PA), transferrin (TR), PaO2, PaCO2, WBC and APACHE Ⅲ score were measured for the patients with 24-hour continuous nutrition (group A) and 16-hour cyclic nutrition (group B)infusion.Effects of the two nutritional therapies were compared.Results After nutrition infusion, serum levels of ALB, PA and TR were (34±3)g/L, (196±28)mg/L and (2.1±0.3 ) g/L in group A, and (35 ±4) g/L, (198 ±25) mg/L and (2.0 ±0.4) g/L in group B, respectively; and PaO2 and PaCO2levels were (92 ± 12) mm Hg ( 1 mm Hg =0.133 kPa) and (42 ± 10) mm Hg in group A, and (91 ±9)mm Hg and (42 ± 10) mm Hg in group B, respectively.WBC and APACHE Ⅲ score were ( 11.8 ± 1.7) ×109/L and 38 ±7 in group A, and ( 12.6 ± 1.2) × 109/L and 40 ±6 in group B, respectively.Significant difference in serum levels of PA and TR was found between the two groups (PPA =0.019 and PTR =0.013),while there was no significant difference in other indictors between the two groups.Conclusions 24-hour continuous nutrition infusion for patients with MVRF can obviously improve their serum levels of PA and TR,but has no effect on serum level of ALB, PaO2, WBC and APACHE Ⅲ score in critical ill patients, as compared to those with 16-hour cyclic nutrition infusion.

This paper aims to investigate the value of diffusiion weighted imaging (DWI) and different apparent diffusion coefficient (ADC) methods to predict the curative effects of neoadjuvant chempotherapy (NAC) for breast cancer. From March 2010 to December 2012, seventy-one patients were pathologically confirmed invasive breast cancer by needle puncture biopsy received before surgery, and underwent magnetic resonance before and after NAC, the ADC were measured by mean ADC method and lower ADC method. The pathologic response after NAC was divided to major histological response (MHR) group and non-major histological response (NMHR) group according to Miller & Payne system. Results displayed that ADC values obtained before NAC, at the end of the second cycle of NAC, and after whole course of treatment, had good correlations between mean and lower ADC methods (the Pearson's correlation=0.699, 0.749 and 0.895, respectively). Significant difference in ADC obtained both with mean and lower ADC methods could be found between MHR and NMHR groups after the second cycle of NAC (P< 0.05). After the second cycle of NAC, significant difference in the change rate of ADC could be found between MHR and NMHR groups by using lower ADC method (P<0.05), but not be found by using mean ADC method (P >0.05). In conclusion, DWI could monitor the pathologic changes of breast cancer after NAC, and the lower ADC method might be used to evaluate the curative effect of NAC with the change rate of ADC.
Breast Neoplasms ; drug therapy ; pathology ; Diffusion Magnetic Resonance Imaging ; Female ; Humans ; Neoadjuvant Therapy

Animals ; Male ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; chemically induced ; Silicon Dioxide ; chemistry ; toxicity

AIMTo establish a solid phase extraction-high performance liquid chromatographic (SPE-HPLC) method for determining plasma scutellarin concentration, and study its pharmacokinetics after i.v. breviscapine in rabbits.

METHODSMethanol-water-phosphoric acid (50:50:0.5) mixture was used as mobile phase, Nucleosil C18 column (250 mm x 4.6 mm ID) was selected. The wavelength of UV detection was 335 nm. Fifteen rabbits, randomized into 3 groups, were given breviscapine i.v. at the dose of 10, 20 and 40 mg.kg-1. Scutellarin in plasma was determined by SPE-HPLC method.

RESULTSLinearity was obtained over the range of 0.02-10.0 mg.L-1 of scutellarin. The method recovery was 96.15%-99.31%; the within-day and between-day RSDs were all below 10%. After i.v. 10, 20 and 40 mg.kg-1 of breviscapine to rabbits, the concentration-time curve of scutellarin fitted to a three compartment model. The main pharmacokinetic parameters showed no significant difference between low and medium doses, but the difference was significant between high dose and other doses.

CONCLUSIONThis assay method was accurate, sensitive, simple and suitable for the measurement of plasma scutellarin concentration. The pharmacokinetic characteristics were found to fit a three-compartment model following i.v. injection of breviscapine to rabbits. The changes of drug concentration in vivo exhibited linear kinetics ove the dosage range of 10-20 mg.kg-1, but when the dosage was 40 mg.kg-1, the linear kinetic properties disappeared.

Animals ; Apigenin ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Flavonoids ; blood ; pharmacokinetics ; Male ; Rabbits

Child ; Humans ; Hydrogen Sulfide ; Pneumonia

Quantification of cytotoxic T lymphocytes (CTL) is extremely important due to the pivotal role they play in controlling pathogen infection and anti-tumor actions. Previously used methods for detecting specific CTL are usually indirect. In recent years, tetramer technology has been developed to directly visualize antigen-specific CTL efficiently, and become the critical approach in studying T cell immune responses. A simplified procedure for preparing tetramers is reported here in this paper and a tetramer loaded with human cytomegalovirus (HCMV) peptide was successfully obtained using this procedure, which possessed binding activity with specific CTL. The heavy chain of HLA-A * 0201 gene was cloned by RT-PCR from HLA-A2+ donor. An expression vector, encoding the extracellular domain of HLA-A * 0201 heavy chain (A2) fused with a BirA substrate peptide (BSP) at its carboxyl terminus, was constructed by PCR with cloned A2 gene as the template. The A2 heavy chain was expressed in Escherichia coli mostly in the form of inclusion body and purified by washing inclusion body. The monomer of soluble A2 loaded with peptide was reconstructed by dilution from the heavy chain in the presence of light chain beta2-microglobulin and HLA-A2 restricted HCMV pp65(495-503) peptide (NLVPMVATV, NLV). Refolded A2-NLV monomer was biotinylated with a commercial BirA and purified by low pressure anion exchange chromatography on a Q-Sepharose (fast flow) column. The tetramer was then formed by mixing A2-NLV monomer with streptavidin-PE in a ratio of 4:0.8 leading to more than 85% multiplication as revealed by SDS-PADE under non-reducing conditions without boiling the sample. Flow cytometry analysis indicated that this tetramer could bind to specific CTL from HLA-A2+ donor. In conclusion, a simplified procedure is established to prepare HLA-A2 tetramer, which may not only facilitate the application of tetramer technology for studying specific T lymphocyte immune response but A2-NLV itself be applied clinically to monitor CMV-specific CTL in stem cell and organ transplantation.
Cloning, Molecular ; Cytomegalovirus ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; HLA-A Antigens ; biosynthesis ; genetics ; immunology ; HLA-A2 Antigen ; Humans ; Phosphoproteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; Viral Matrix Proteins ; biosynthesis ; genetics

Human beta2-microglobulin (beta2m) is the light chain of major histocompatibility complex (MHC) class I molecule. High-yield production of this protein is a prerequisite to the preparation of MHC I tetramer. The present study aims to obtain recombinant human beta2m expressed in Escherichia coli (E. coli), for the purpose of preparing MHC class I tetramers. For cloning of human beta2m gene, a pair of specific primers was designed based on the published sequence of this gene and the cDNA of full coding region for beta2m precursor was obtained by RT-PCR from the total RNA of human leukocytes. The amplified cDNA was subsequently cloned and its sequence was confirmed by DNA sequencing analysis (the sequence has been deposited in GenBank with accession number of AY187687). The prokaryotic expression vector containing a gene encoding mature beta2m was constructed by inserting the DNA fragment, which was generated by PCR reaction with the cloned beta2m gene as template, into an IPTG-inducible expression vector pET-3c plasmid. The first eight codons for N terminal amino acid residues of beta2m were optimized for its expression in E. coli. The complete sequence of beta2 m gene in the expression vector was verified by DNA sequencing analysis. High-yield expression of beta2m was achieved in E. coli transformed with the expression vector, and most of the recombinant beta2m existed in the inclusion body after IPTG induction. The inclusion body was washed extensively and beta2m in the inclusion body was solublized with 8 mol/L urea. The beta2m was refolded by dialysis and purified by ion-exchange chromatography (Q-Sepharose). Western blotting assay indicated that the polyclonal antibody against human native beta2m could react specifically with the recombinant protein. The purified protein appeared as a single band on both SDS-PAGE and Western blotting, indicating that it was chemical and antigenic pure. This work establishes a convenient approach for renaturation and purification of large quantity of recombinant beta2m which is identical to the native protein without any tags fused except for a methionine residue at the amino terminus. This provides the basis for the preparation of MHC tetramers.
Base Sequence ; Blotting, Western ; Cloning, Molecular ; Escherichia coli ; genetics ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; biosynthesis ; chemistry ; immunology ; beta 2-Microglobulin ; chemistry ; genetics ; immunology

OBJECTIVETo compare the therapeutic effects of opposing needling and routine acupuncture for treatment of poststroke shoulder-hand syndrome.

METHODSSixty cases were randomly divided into an opposing needling group and a routine acupuncture group, 30 cases in each group. In the two groups, Jianyu (LI 15), Jianliao (TE 14), Quchi (LI 11), Hegu (LI 4), etc. were selected, with on the healthy side selected for the opposing needling group and on the affected side selected for the routine acupuncture group. The motor function of the affected limb was evaluated by Fugl-Meyer scale, activity of daily living by ADL scale, and pain by VAS, and the edema degree was investigated before and after treatment.

RESULTSThere were significant differences before treatment and after 2 therapeutic courses in the scores of Fugl-Meyer for the upper limb motor function and the ADL score for activity of daily living (both P<0.05), with the opposing needling group being significantly better than the routine acupuncture group (P<0.05); after treatment, both the edema degree and the pain scores significantly decreased (both P<0.05), with the opposing needling group in improvement of the edema degree being better the routine acupuncture group (P<0.05), and with no significant difference between the two groups in improvement of pain (P>0.05); the total effective rate was 93.3% in the opposing needling group and 90.0% in the routine acupuncture group with a significant difference between the two groups (P<0.05).

CONCLUSIONThe clinical therapeutic effect of the opposing needling is better than that of routine acupuncture therapy for treatment of poststroke shoulder hand syndrome.

Acupuncture Points ; Acupuncture Therapy ; methods ; Aged ; Female ; Humans ; Male ; Middle Aged ; Needles ; Reflex Sympathetic Dystrophy ; etiology ; therapy ; Stroke ; complications

OBJECTIVETo distinguish the edema, injury, or rupture in the traumatic skeletal muscle fiber in vivo using diffusion tensor imaging (DTI) and tractography on magnetic resonance imaging (MRI).

METHODSThe skeletal muscle trauma models were made in 4 rabbits (eight hindlimbs) by iron discus (weight 1.0 kg, diameter 6 cm) falling down vertically from 45 cm height to rabbits' thighs. Conventional sequences and two-dimensional (2D) diffusion-weighted (DW) spin-echo (SE) echo planar imaging (EPI) sequence with fat suppression (b = 600 s/mm2) were performed on 1. 5T MRI scanner. The grading of edema, injury, and fiber rupture in the damaged muscle were made according to their histopathological views, which was consistent with the images. The mean apparent diffusion coefficient (ADC) values and fractional anisotropy (FA) values were measured from the region of interests (ROIs) of all groups on 2D DW images used for tractography. Analysis of variance test was performed to analyze all data.

RESULTSADC values of the areas in normal muscle, edema muscle, injury muscle, and ruptured muscle were (6.12 +/- 1.34) x 10(-3), (6.38 +/- 1.30) x 10(-3), (8.06 +/- 0.97) x 10(-3), and (9.57 +/- 0.93) x 10(-3) mm2/s, respectively. There was significant difference among groups (P < 0.001), but no difference between edema muscle and normal muscle group (P > 0.05). The FA values of normal muscle, edema muscle, injury muscle, and ruptured muscle were 0.42 +/- 0.12, 0.36 +/- 0.12, 0.26 +/- 0.09, 0.12 +/- 0.08, respectively, with a significant difference among groups (P < 0.001). In the edema muscle, the tracking cross-fiber could be seen but it decreased slightly. In the injury muscle, the tracking fiber decreased markedly. In the ruptured muscle, the transverse-orientation tracking fiber vanished, yet some interrupted longitudinal-orientation tracking fiber could be found.

CONCLUSIONThe edema, injury, and rupture of muscle fiber in rabbit damaged skeletal muscle can be verified according to the ADC and the FA on DTI and tractography.

Animals ; Anisotropy ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; Edema ; diagnosis ; pathology ; Male ; Muscle, Skeletal ; injuries ; pathology ; Rabbits ; Rupture ; diagnosis ; pathology ; Thigh ; injuries ; pathology