Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals ; Body Fluids ; chemistry ; Chemical Precipitation ; Chemistry Techniques, Analytical ; methods ; Humans ; Liquid-Liquid Extraction ; Proteins ; isolation & purification ; Solid Phase Extraction

Hepatitis B is one of the most serious global threats to human health. Phylogenetic analysis of hepatitis B virus (HBV) can reveal the evolutionary relationship between HBV sequences and thus provide a basis for the prediction and treatment of hepatitis B and other aspects. In this study, we performed sequence analyses on the HBV sequences of five clinical HBV samples and the HBV sequences retrieved from the GenBank, EMBL, and DDBJ to construct a phylogenetic tree and analyze sequence structures. The experimental results revealed that the C gene of one cloned sequence had a recombinant structure of HBV B/ C subtype. Moreover, the phylogenetic results proved the existence of a newly found subtype HBV/B6 in Xishuangbanna of Yunnan Province, China. The experimental conclusion represents certain value for phylogenetic studies of HBV in Yunnan ethnic minority groups.
DNA, Recombinant ; genetics ; Genes, Viral ; genetics ; Genotyping Techniques ; Hepatitis B virus ; classification ; genetics ; Humans ; Phylogeny

Objective To evaluate the value of fMRI guided brain surgery for the lesions in or around Broca's area.Methods Forty-three patients with lesions in or adjacent to the Broca's area were studied.fMRI imaging was obtained by BOLD technique with the tasks of reciting.Fiber tract imaging of white matter was obtained by DTI technique.All functional imaging and anatomic imaging were transferred to neuronavigation system.The technique of direct cortical stimulation was used to validate the language cortex in fMRI.The lesions were resected in microscope.Results Broca's area activation was detected in 38 cases..The distance between the fMRI peak and direct cortical stimulation was rated as overlapping (<1 cm diatance) in 25 cases and neighbouring (<2 cm diatance) in 11 cases.Total lesion resection was achieved in 17 cases, subtotal resection in 14 cases, and partial resection in 12 cases.Postoperative neurological functions were improved in 8 cases, unchanged in 31 cases, and temporary worsen in 4 cases.Conclusions The identification of the Broca's area by reciting task in fMRI is sensitive and precise.The fMRI is helpful to decrease the side effect injury in the brain surgery for the lesions in or around the Broca's area.

Objective To explore the imaging features of papillary thyroid microcarcinoma(PTMC) with real time contrast-enhanced ultrasound.Methods One hundredand forty-three cases with 149 thyroid nodules(no diffuse lession) were divided into two groups according to the diameter size(group 1,＜0.5 cm;group 2,0.5-1.0 cm) and examined by contrast-enhanced ultrasound during preoperation.Pathology was followed up as golden diagnosis criteria.Results Seventy-five benign tumors and 74 PTMC were confirmed by pathology.There were significant differences in echoes homogeneity between benign and malignant tumors in group 2(W =1 029.5,Z =-5.524,P =0.000) but no in group 1(W =933.0,Z =-1.738,P =0.082).And nonhomogeneous enhancement were showed in most PTMC in group 2.But most PTMC showed homogeneous enhancement in group 1.Conclusions Contrast-enhanced ultrasound is valuable in diagnosis of PTMC with the diameter size of 0.5-1.0 cm.

Objective To develop an efficient and rapid method to synthesize 68 Ga?DOTA?TATE, and to perform PET/CT imaging on neuroendocrine tumor (NET) bearing mice and one patient. Methods 68GaCl3 was eluted by 0.05 mol/L HCl in 68 Ge?68 Ga generator, then added to pre?adjusted DOTA?TATE buffer, and heated for 15 min at 85 ℃, then filtered with 0.22 μm microfiltration membrane. The radiochemical purity and in vitro stability of 68 Ga?DOTA?TATE were analyzed with radio?HPLC. The biodistribution and PET/CT imaging on HT?29 colon cancer xenografts mice model were performed. After approved by Peking University Cancer Hospital Ethics Committee, a case of NET patient received 148 MBq 68 Ga?DOTA?TATE intravenous?ly, followed by a whole?body scan at 60 min post?injection. Results 68 Ga?DOTA?TATE was prepared in high radiochemical purity (>98%) and with specific activity of 55 GBq/μmol in 25 min. The tracer showed excellent in vitro stability and no acute toxicity. PET/CT imaging on mice model showed positive uptake in tumor tissues. PET/CT imaging on one NET patient showed positive uptake of 68 Ga?DOTA?TATE in primary and metastases lesions. Conclusions A rapid, high radiolabeling yield and high specific activity method to prepare 68 Ga?DOTA?TATE is established. 68 Ga?DOTA?TATE could be specifically uptaken by somatostatin receptor ( SSTR) positive NET with high affinity. It could serve as a useful tool for early diagnosis of SSTR positive NET and has clinical prospective for somatostatin?mediated targeting therapy.

Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

ObjectiveTo apply the Schwann cell neurilemma channel(SCNC) in promoting the regeneration of injuried spinal cord.MethodsSpinal cords of rats after laminectomy at T8/9 were divided in 3 groups randomly: 10 rats were transplanted with SCNC(Group A);10 rats only made models(Group B);10 rats were as normal control group(Group C).All rats received behaviour survey and inclined plane test every week 8 weeks after operation.6 weeks after operation,they were measured with cortical somatosensory evoked potential(CSEP).And 8 weeks after operation,all specimens were studied histologically.ResultsCSEP could be recorded in group A and C,but not in group B.There were marked differenties between group A or C and group B in behavior survey,inclined plane test.Amount of nerve fibers regenerated through the injury gap in group A was significantly larger than that in group B.ConclusionSCNC could facilitate axonal regeneration and promote the repairing of injuried spinal cord.

OBJECTIVETo investigate the potential role of serum citrulline level in evaluating the intestinal absorptive area and capacity in patients with short bowel syndrome (SBS).

METHODSSerum citrulline concentration was determined using high performance liquid chromatography (HPLC) in SBS patients (n=22) and healthy controls (n=33). In SBS patients, the remnant small bowel lengths and diameters were measured by radiography, and their 5- hour urine D- xylose excretion and intestinal protein absorption were also determined. The correlationship of serum citrulline level with remnant small bowel length, surface area, protein and D- xylose absorption was analyzed. The 6 patients receiving intestinal rehabilitative therapy, serum citrulline level, protein and D- xylose absorption after therapy were also measured.

RESULTSSerum citrulline level of SBS patients was significantly lower than that of healthy controls [(5.94+/- 2.65) vs [(16.87 +/- 5.97) micromol/L, P < 0.01]. In SBS patients, serum citrulline was positively correlated with remnant small bowel length and surface area (r=0.82 and r=0.86 respectively). There was also a significant correlationship of serum citrulline level with 5- hour D- xylose excretion (r=0.56) and intestinal protein absorption (r=0.48). Serum citrulline, 5- hour D- xylose excretion and intestinal protein absorption were all significantly raised in patients after rehabilitative therapy, although no correlation of increasing percentage was found among above three parameters.

CONCLUSIONSSerum citrulline concentration is positively correlated with intestinal absorptive area and capacity in SBS patients. It is a potential marker for evaluating the severity of intestinal failure and the efficacy of rehabilitative therapy in short bowel patients.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Citrulline ; blood ; Female ; Humans ; Intestine, Small ; metabolism ; Male ; Middle Aged ; Short Bowel Syndrome ; blood ; metabolism ; physiopathology ; Xylose ; metabolism ; Young Adult

A new method for simultaneous determination of six benzoic acid esters and their metabolite p-hydroxybenzoic acid (p-HB) in sediments was developed.The samples were extracted with methanol and water,and then the extraction was purified by Oasis MCX solid phase column.The separation of targets was conducted by passing through a UPLC?HSS T3 column with a methanol-0.1% acid aqueous solution as the mobile phase under gradient elution.The qualitative analysis of the analytes was measured by Ultra performance liquid chromatography-Triple quadrupole tandem mass under multiple reaction monitoring mode (MRM) and the internal standard method was applied for quantitative analysis.The results indicated that seven target substances had a good linear relationship in the range of 0.1-500 μg/L with correlation coefficients above 0.9995 and the detection limits of 0.75-1.23 ng/g.The recoveries ranged from 65.5% to 88.3% and the relative standard deviation was 7.4%-13.6%.This method was suitable for the measurement of benzoic acid esters with the advantages such as simple processing,good selectivity and high sensitivity.

OBJECTIVETo study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis, so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree.

METHODSForty-four Japanese healthy 6-month-old rabbits (equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method. The rabbits were randomly divided into 6 groups: normal group, model group, control group (microwave), experimental group 1 (electricity), experimental group 2 (thermal), experimental group 3 (kneepad). Ten rabbits in the normal group were breed with conventional method; 9 rabbits in the model group were breed with conventional method after model made; 9 rabbits in the control group were treated with microwave for 30 minutes, one time daily; 9 rabbits in the experimental group 1 were treated with electricity (density wave) for 30 minutes,one time daily;8 rabbits in the experimental group 2 were treated with hot (hot soft membrane) for 30 minutes, one time daily; 9 rabbits in the experiment group 3 were treated with electrothermal (OA knee pad) for 30 minutes, one time daily. All the rabbits were treated for 16 weeks and then sacrificed. The expressions of Bcl-2 and p53 mRNA of chondrocytes in knee joint were detected by using fluorescence quantitative RT-PCR method.

RESULTSAt the 16 hthek,th e OD260/OD280 value range of total RNA extracted from rabbit articular cartilage tissue in each group were all at 1.80 to 2.00,wh ich indicates high RNA purity. The p53 relative mRNA in articular cartilage cells of model group,th e control group,th e experimental group 1 ,r oup 2,gr oup 3 were overexpressed,an d Belc2 mRNA expression levels of articular cartilage cells were low expression,an d compared with the normal group there were significant differences (P < 0.01). Belc2, p53 mRNA expression in articular cartilage cells,th ere were significant differences (P < 0.01) between the control group, experimental group 1, group 2, group 3 and model group. The results between the control group, experimental group 1 ,group 2 and group 3 had significant differences (P < 0.01).

CONCLUSIONOA-kneepad can up-regulate the mRNA expression of Bcl-2 as well as down-regulate the mRNA expression of p53, thereby to inhibit the apoptosis of cartilage cells and delay the degeneration of articular cartilage changes.

Animals ; Apoptosis ; physiology ; Disease Models, Animal ; Female ; Knee Joint ; metabolism ; pathology ; Male ; Osteoarthritis, Knee ; genetics ; pathology ; therapy ; Protective Devices ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Rabbits ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics