Objective To construct eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG,and to detect the expression of the plasmid in vitro.Methods hOPG cDNA fragment was extracted from plasmid MGC:29565,amplified by PCR,and inserted into pcDNA3.1(-)vector,then identified by restriction endonuclease digestion and nucleotide sequencing.C2C12 cells were transfected with the plasmid using Lipofectamine 2000.The expression of OPG protein was detected by immunohistochemistry techniques and bone wafer pit assay.Results The constructed recombinant plasmid contained the sequence of hOPG gene.After transfection with the plasmid,active OPG protein could be expressed in C2C12 cells.Conclusion The constructed eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG can express OPG in vitro,which is the basis for further study on the treatment of bone absorption caused by abnormal activity of osteoclasts.

OBJECTIVETo explore methods and clinical outcomes of microsurgical technique in treating patients with severed palm.

METHODSFrom January 2009 to December 2012,45 patients with severed palm were treated by replantation through microsurgical technique, included 37 males and 8 females, aged from 13 to 45 years old with an average of 25. Postoperative survival rate and evaluation standard of upper limb replantation function posposed by Chinese Medical Association were applied for evaluate clinical outcomes after operation.

RESULTSForty-five patients with severed plam were treated by replantation. Thirty-nine patients (121 fingers) were survived,and survival rate was 87%. All patients were followed up 3 to 15.5 months with an average of 11.5 months. According to evaluation standard of upper limb replantation function posposed by Chinese Medical Association, the total score was 80.27 +/- 1.93, and 27 cases got excellent results, 8 good and 4 poor.

CONCLUSIONThe success of replantation of severed palm depends on grasping operation indication strictly, knowing complexity and local anatomic relationship, debridement completely during operation, making full use of remaining organization, arteriovenostomy through microsurgical technique as early as possible, constructing circulation and repairing injuried nerve rationally.

Adolescent ; Adult ; Female ; Hand Injuries ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Replantation ; Young Adult

The shape and structure of granules are controlled by the granulation process, which is one of the main factors to determine the nature of the solid dosage forms. In this article, three kinds of granules of a traditional Chinese medicine for improving appetite and promoting digestion, namely, Jianwei Granules, were prepared using granulation technologies as pendular granulation, high speed stirring granulation, and fluidized bed granulation and the powder properties of them were investigated. Meanwhile, synchrotron radiation X-ray computed micro tomography (SR-µCT) was applied to quantitatively determine the irregular internal structures of the granules. The three-dimensional (3D) structure models were obtained by 3D reconstruction, which were more accurately to characterize the three-dimensional structures of the particles through the quantitative data. The models were also used to quantitatively compare the structural differences of granules prepared by different granulation processes with the same formula, so as to characterize how the production process plays a role in the pharmaceutical behaviors of the granules. To focus on the irregularity of the particle structure, the box counting method was used to calculate the fractal dimensions of the granules. The results showed that the fractal dimension is more sensitive to reflect the minor differences in the structure features than the conventional parameters, and capable to specifically distinct granules in structure. It is proved that the fractal dimension could quantitatively characterize the structural information of irregular granules. It is the first time suggested by our research that the fractal dimension difference (Df,c) between two fractal dimension parameters, namely, the volume matrix fractal dimension and the surface matrix fractal dimension, is a new index to characterize granules with irregular structures and evaluate the effects of production processes on the structures of granules as a new indicator for the granulating process control and optimization.
Drugs, Chinese Herbal ; analysis ; Fractals ; Medicine, Chinese Traditional ; Powders ; Quantitative Structure-Activity Relationship ; Synchrotrons ; Technology, Pharmaceutical ; Tomography, X-Ray Computed

A method of comprehensive chemical pattern recognition of Atractylodis Rhizoma was established by GC-MS fingerprint, principal component analysis, cluster analysis and discriminant analysis. A DB-wax column (0.25 mm x 60 m, 0.25 microm) with El ion source and 70 V electron multiplier were used for GC-MS analysis. Using principal component analysis, cluster analysis, and discriminant analysis, 15 common peaks of sample fingerprints for chemical pattern recognition research were analysed. The same results were obtained from the fingerprint, principal component analysis and cluster analysis, which could use to distinguish genuine Atractylodes lancea, ungenuine A. lancea and A. chinensis. Thus, this method could be used for the quality control and comprehensive evaluation of Atractylodis Rhizoma.
Atractylodes ; chemistry ; China ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Quality Control ; Rhizome ; chemistry

Objective Application of the second metatarpophalangeal joint by traction prolong trans- plant repair the defects in the metacarpophalangeal joint,reconstruct the function of it.Methods By means of the apparatus to prolong finger in advance,then transplant the second metatarpophalangeal joint to recon- struct metacarpophalangeal joint for seven cases of obsolete defects in the metacarpophalangeal joint.Results The average of finger prolong was 2.6 cm,consultation from 1 to 4 years.average 2.5 years,thai the trans- plant joints have all survived and osteal concrescence.Through the criterion Chinese Medical Association,good rate was 85.7%. Conclusion It' s a good method to repair obsolete defects in the metacarpophalangeal joint by transplant traction prolong of the second metatarpophalangeal joint.

OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.

METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.

RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.

CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome

BACKGROUND: The thermo-sensitive composite hydrogels have gained increasing interest in bone regeneration domain due to their biomimetic extracellular matrix (ECM) structure, good biocompatibility, minimal invasive performance and in situ molding. OBJECTIVE:To prepare a thermo-sensitive injectable alginate/α-tricalcium phosphate (α-TCP)/collagen I (Alg/TCP/Col) composite hydrogel and explore its characterization. METHODS:Ca-carrying interdigitation-fusion vesicles (Ca-IFVs) were prepared. The liposomes carrying the optimal concentration of calcium ions were selected for the following experiments by investigating their encapsulation efficiency and drug loading rate. Alg/TCP/Col precursor solution (Alg or Alg/TCP precursor solution) was mixed with Ca-IFVs at 37℃ in different proportions (5, 10, 15, 20) to prepare thermosensitive hydrogels. The structure, rheology behavior, volume swelling ratio, and mechanical properties of the composite hydrogel were observed. MC3T3-E1 cells were co-cultured with Alg/TCP/Col, Alg, and Alg/TCP hydrogels, respectively. Then, morphology of the cells was observed by confocal microscopy at 1, 3, 7 days after co-culture. RESULTS AND CONCLUSION:(1) The pore size of the freeze-dried hydrogel was 50-100 μm, and TCP particles uniformly adhered to the surface of the Alg/TCP hydrogel surface. The Alg/TCP/Col hydrogel was a dense aggregate with collagen fibers in contrast to the Alg/TCP hydrogel. (2) The Alg/TCP/Col hydrogel exhibited a suitable phase transition temperature (Tm) between 35-39℃. (3) The volume swelling ratio of the hydrogel was increased with the increase of Ca-IFVs size. When the α-TCP complex was added into the Alg/TCP hydrogel, the swelling ratio decreased slightly. Alg/TCP/Col hydrogel exhibited a higher swelling ratio than the Alg/TCP hydrogel. (4) When the mixture ratio of precursor solution to liposome was 10, the compressive modulus of Alg/TCP/Col hydrogel and Alg/TCP hydrogel was significantly higher than that of the Alg hydrogel (P<0.05). (5) When the mixture ratio of precursor solution to liposome was 10, round MC3T3-E1 cells were observed on the Alg hydrogel; the cells on the surface of the Alg/TCP hydrogel were scattered and tended to extend; the cells on the surface of the Alg/TCP/Col hydrogel had a stress-extended morphology, and grew into the hydrogel, and meanwhile, the cell number increased significantly. To conclude, the liposome-mediated Alg/TCP/Col has good mechanical properties and cytocompatibility.

Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.

[Objective]To investigate the effect of GRACE scores on prediction of 30-day cardiovascular adverse events in acute chest pain patients.[Methods]A prospective,observational analysis was conducted in the patients with acute chest pain in Emergency Department(ED)from January 1,2016 through April 1,2016.Data including characteris-tics and GRACE scores were collected.All causes leading to MACE were followed up at 30th day after the onset of acute chest pain.[Results]Among a total of 209 patients presenting with acute chest pain enrolled in this study,110 were male (52.63%)and 99 were female(47.37%).The range of age was 20-98years old,and mean age was(65.28±16.85)years old.During follow-up period,12 patients had MACE,2 patients died in ED,3 patients died in hospital,6 patients died out of hospital,and 1 person was diagnosed with myocardial infarction. When compared with non-MACE group,factors including age,BMI,hospitalized patient number,and number of patients admitted in CCU as well as GRACE scores, were significantly higher in MACE group(P<0.05). The predictive ROC curve area of GRACE scores in 30-day MACE was 0.819(0.735 to 0.902). The optimal sensitivity and specificity were 0.92 and 0.65,respectively. The probability of 30-day cardiovascular adverse events in various GRACE score risk stratification was 0.95%(low-risk),6.67%(medi-um-risk),and 18.92%(high-risk),respectively.[Conclusion]The GRACE score was a useful predictor to the occur-rence of 30-day cardiovascular adverse events in acute chest pain patients.

Objective To identify the ABCD1 gene mutation in a patient suspected with adrenoleukodystrophy (ALD) and perform its gene analysis in his family members to make a definite diagnosis. Methods Total RNA and genomic DNA were extracted from the leukocytes of peripheral blood in the proband and the family members. The ABCD1 coding region of cDNA in the proband was amplified and sequenced. Mutational site in the ABCD1 gene of the proband was further confirmed by PGR and direct sequencing; at the same time, the mutation in the ABCD1 gene of the genomic DNA in the family members was analyzed by direct sequencing. Results Two bases deletions (656_657delGA)were identified and the corresponding mutation of fs R89 was detected in the ABCD1 gene of the proband, which could make the definite diagnosis of ALD that belonged to adrenomyeloneuropathy. The same gene mutation (ALD hemizygote) was noted in his cousin; his mother, younger sister of his mother and his younger female cousin were noted as the ALD carrers. His older sister was noted as ABCD1 normal genotype. Conclusions A novel ABCD1 gene mutation (fs R89) was identified in Chinese patient with ALD. Molecular testing is an effective way in making diagnosis on patient suspected as having X-linked ALD.