To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To set up a method of stools protein extraction,analysis and identification in order to get the new nonin-vasive indicators of human digestive diseases.Methods The stools proteins,collected from healthy persons,the patients with atrophic gastritis,those who suffed from gastric carcinoma and postoperative patients with gastric carcinoma respectively, were extracted in three different ways including saline,Tris-HCl buffer and Urea buffer,the best way was selected by using SDS-PAGE,then a preliminary analysis of stools proteins was performed by 1D LC-MS/MS.Results The methods of saline and Tris-HCl buffer could get more stools proteins than the method of urea.The proteins in stools from the healthy persons, the patients with atrophic gastritis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma were all abundant and more than one hundred.There was a significant difference in stools protein maps among the various populations.Alpha1-antitrypsin,a number of immunoglobulin and keratin were identified in the stools from patients with gastric carcinoma but not postoperative patients with gastric carcinoma and the healthy persons.Conclusions In this re-search,there was a significant difference in stools protein maps among the healthy persons,the patients with atrophic gastri-tis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma,not only the composition of stools proteins,but also the abundance of same proteins.Therefore,using proteomics technologies to screening of the noninvasive indicators in human stools is viable.The study recommended that the noninvasive indicators in human stools should be iden-tified with quantitative differences analysis combination of quality of mass spectrometer method in the future research.

Objective To evaluate the swallowing function of patients with dysphagia after brainstem injury using manofluorography (MFG),analyzing the abnormal biomechanical and kinematic parameters as well as any correlation between changes in the pharynx and the upper esophageal sphincter (UES) measured manometrically and changes in the kinematics of the hyoid bone.Methods Thirteen patients with dysphagia after brainstem injury (the patient group) and 13 healthy participants (the control group) underwent manofluorography.Kinematics and biomechanical changes during swallowing were compared between the two groups and the correlations between the observations were analyzed.Results The patient group showed significantly lower maximum pressure and rate of pressure change at the base of the tongue and in the hypopharynx,as well as less hyoid anterior displacement,smaller and briefer UES opening,but significantly higher minimum pressure at UES relaxation.The duration of tongue root elevation and hypopharynx pressure was also shorter than in the control group,on average.There was a negative correlation between hyoid anterior displacement and the minimum pressure on UES relaxation in the control group,and a positive correlation between hyoid anterior displacement and the maximum pressure at the base of the tongue and in the hypopharynx in the patient group.Conclusions The concurrent use of manometry and video-fluorography for evaluating dysphagia can be an objective and effective diagnostic tool for the comprehensive evaluation of swallowing function.

OBJECTIVETo observe the clinical efficacy of Tongfu Mixture (TM) for post-ERCP pancreatitis (PEP).

METHODSTotally 54 PEP patients were randomly assigned to the control group (treated by routine therapy, 26 cases) and the TM treatment group (treated by TM, 28 cases). Clinical indices including the alleviation time of abdominal pain/distention, gastrointestinal function recovery time, and the post-surgical length of stay were observed. Blood amylase (AMY), C-reactive protein (CRP), plasma endotoxin (PLS), TNF-alpha, and IL-6 were detected before surgery, 12 h, 48 h, and 96 h after surgery.

RESULTSThe alleviation time of abdominal pain/distention, the gastrointestinal function recovery time, and the post-surgical length of stay were obviously shorter in the TM treatment group than those in the control group (P < 0.05). The recovery of AMY and CRP were better in the TM treatment group than in the control group at post-operative 48 h and 96 h (P < 0.05). The levels of LPS, TNF-alpha, and IL-6 were lower in the TM group than in the control group at post-operative 96 h (P < 0.05).

CONCLUSIONTM showed better clinical efficacy and could significantly decrease the post-surgical length of stay. post-ERCP pancreatitis; integrative medicine; Tongfu Mixture

Adult ; Cholangiopancreatography, Endoscopic Retrograde ; adverse effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; drug therapy ; etiology ; Phytotherapy

OBJECTIVETo investigate surgical procedure and clinical effects for the treatment of patellofemoral osteoarthritis (PFOA) with arthroscopic patelloplasty and circumpatellar denervation.

METHODSTotally 156 patients with PFOA (62 males and 94 females; aged from 45 to 61 years old with an average of 52) were treated from September 2011 and March 2013. The clinical manifestations included recurrent swelling and pains on knee-joint,and aggravated pains upon going up/down-stairs, squatting-down, or standing-up. PFOAs were treated with arthroscopic patelloplasty and circumpatellar denervation. The therapeutic effects before and after surgery were evaluated by Lysholm scores and Kujala scores. The therapeutic effects were graded by classification of cartilage injury degrees.

RESULTSTotally 149 cases were successfully followed up for 14.8 months. The incisions were healed well and no complication were occurred. Lysholm score was improved from 73.29 preoperatively to 80.93 postoperatively and average Kajala score from 68.34 preoperatively to 76.48 after surgery. This procedure was highly effective for patients with I - III degree cartilage injury, while not for patients with IV degree cartilage injury.

CONCLUSIONFor PFOA patients, arthroscopic patelloplasty and circumpatellar denervation is effective for significantly relieving anterior knee pains, improving knee-joint functions and quality of life, and deferring arthritic procession.

Arthroscopy ; Denervation ; Female ; Humans ; Knee Joint ; innervation ; surgery ; Male ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; surgery ; Range of Motion, Articular ; Treatment Outcome

OBJECTIVETo explore causes of gluteal muscle contracture induced anterior knee pain and curative effect of arthroscopic release.

METHODSFrom March 2002 to August 2013,36 patients with gluteal muscle contracture induced anterior knee pain were treated, including 15 males, 21 females, aged from 9 to 40 years old with an average (18.7±7.2) years old; the courses of diseases ranged from 4 to 30 years. The clinical manifestations involved limited to symmelia, positive Ober sign, buttocks touch contracture belts, knee and patella slide to lateral when doing squat activities. All patients were performed gluteal muscle contracture release under arthroscopic. Postoperative complications were observed, Kujala scoring before and after operation was used for compare curative effect.

RESULTSAll patients were followed up with an average of 29 months. The incision were healed well, and no complications were occurred. Postoperative Kujala score were improved more than preoperative.

CONCLUSIONGluteal muscle contracture release could alleviate hypertension of lateral patella, and palys an important role in preventing patellofemoral arthritis.

Adolescent ; Adult ; Buttocks ; Child ; Contracture ; physiopathology ; Female ; Humans ; Knee ; Male ; Pain ; etiology

Based on the principle of the pathway engineering, a novel pathway of producing glycerol was built in E. coli. The gpd1 gene encoding glycerol 3-phosphate dehydrogenase and the hor2 gene encoding glycerol 3-phosphatase were cloned from Saccharomyces cerevisiae, respectively. The two genes were inserted into expression vector pSE380 together. A recombinant plasmid pSE-gpd1-hor2 containing polycistron was constructed under the control of the strong trc promoter. Then it was transformed into E. coli BL21. The result showed the recombinant microorganism GxB-gh could convert glucose to glycerol directly. And the recombinant microorganism GxB-gh was incubated to produce glycerol from D-glucose in the fermentor. The maximal concentration of glycerol was 46.67g/L at 26h. Conversion rate of glucose was 42.87%. The study is about "green" producing glycerol by recombinant microorganism and is also useful for further working in recombining microorganism of producing 1,3-propanediol.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Genetic Engineering ; Glycerol ; metabolism ; Glycerolphosphate Dehydrogenase ; biosynthesis ; genetics ; Phosphoric Monoester Hydrolases ; biosynthesis ; genetics ; Saccharomyces cerevisiae ; enzymology ; genetics

OBJECTIVETo explore the effectiveness of hamstring tendon graft embedding reconstruction of medial patellofemoral ligament for the treatment of recurrent dislocation of the patella.

METHODSFrom March 2008 to June 2013,67 patients with recurrent dislocation of patella were treated, including 28 males, 39 females with an average age of 22 years ranging from 10 to 42 years old. The clinical manifestations involved knee joint instability,"giving way", sense of patellar dislocation, anterior knee pain. All 67 patients underwent arthroscopic hamstring tendon graft embedding reconstruction of the medial patellofemoral ligament. The curative effect were evaluated by preoperative and postoperative Lysholm score and Q angle changes.

RESULTSAll cases were followed up after operation and the mean follow-up time was(27.5±13.4) months(4 to 69 months). Postoperative incision were healed well and no patellar dislocation or subluxation occurred. Lysholm score improved from preoperative 76.35±2.86 to 82.71±2.29 postoperatively; Q angle decreased from preoperative (18.75±2.33)° to postoperative(13.28±1.75)°.

CONCLUSIONThe method for the reconstruction of medial patellofemoral ligament can provide enough tension, patellar stability.imoroving the function of knee ioint.

Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Patellar Dislocation ; surgery ; Patellar Ligament ; surgery ; Patellofemoral Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Recurrence ; Tendons ; transplantation

1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.

AIM:To investigate the distribution characteristics of interleukin-22(IL-22)gene rs2227485C/T and rs2227491A/G polymorphisms in Guangxi people and the distribution differences with other ethnic groups ,and to ex-plore the difference levels of common lipid indexes in different genotypes.METHODS:SNaPshot technique and DNA se-quencing were used in 280 Guangxi persons to examine IL-22 genotypes and to analyzed the distribution frequencies of allele and genotype in these sites.The distribution frequencies in different sexes ,and the differences between groups and diffe-rence levels of common lipid indexes in different genotypes were analyzed statistically.RESULTS:Three genotypes of CC ,CT and TT were found in rs2227485C/T with the frequency distribution of 17.1％,49.3％and 33.6％,respectively.No significant difference between different sexes of each genotype and allele frequency in the Guangxi population was observed(P>0.05).Compared with the distribution frequencies of genotype and allele in HapMap -TSI,HapMap-HCB,HapMap-JPT and HapMap-MEX,those in Guangxi population showed statistically significant differences(P<0.05).Three geno-types of AA,AG and GG were found in rs2227491A/G with the frequency distribution of 16.1％,52.8％and 31.1％,re-spectively.There was no significant difference between different sexes of each genotype and allele frequency in the Guangxi population(P>0.05).The significant differences of genotype frequencies among Guangxi population ,HapMap-TSI,Hap- Map-JPT and HapMap-MEX were detected(P <0.05 ).Compared with the other 4 populations ,allele frequencies in Guangxi population had significant difference(P <0.05).There were significant differences in the levels of HDL-C and LDL-C among the 3 genotypes of rs2227491A/G.The level of HDL-C had difference between AG/AA genotype and GG genotype.In addition,the level of LDL-C had difference between AG/GG genotype and AA genotype(P<0.05).CON-CLUSION:rs2227485C/T and rs2227491A/G polymorphisms of IL-22 gene have differences in different populations.The rs2227491A/G polymorphism may be associated with serum lipid levels.