Objective: To construct subtracted cDNA libraries of stomach tumors and normal stomach tissue using suppression subtractive hybridization(SSH).Methods: cDNA Library subtraction was performed using the protocol described in the Clontech PCR-Select cDNA Subtraction Kit. cDNA was synthesized from 2 μg of poly A+RNA from the tumor and normal tissues using AMV reverse transcriptase. The tester and driver cDNAs were digested with RsaⅠ, a four-base-cutting restriction enzyme that yields blunt ends. The tester cDNA was then subdivided into two portions, and each was ligated with different cDNA adaptor. Two hybridizations were performed. In the first, an excess of driver was added to each sample of tester. Hybridization kinetics led to equalization and enrichment of differentially expressed sequences. During the second hybridization, the two primary hybridization samples were mixed together without denaturing and thus the templates were generated from differentially expressed sequences for PCR amplification. Using suppression PCR, only differentially expressed sequences were amplified exponentially and after second PCR amplification the background was reduced and differentially expressed sequences were further enriched. The cDNAs were then directly inserted into a T/A cloning vector to generate a stomach tumor-specific subtracted cDNA library. Results: The amplified library contained 800 positive clones. Plasmid inserts were PCR amplified and showed 250－700 bp inserts. Conclusions: The successfully constructed subtracted cDNA library of gastrocarcinoma and normal tissue enables us to compare two populations of mRNA and obtain clones of genes that expressed in one population but not in the other.The characterization of these genes will allow them to be exploited for their diagnostic and therapeutic potentials.

Objective To establish the model of P2 peptide-induced experimental autoimmune neuritis(EAN)in rats and explore the roles of Th_1/Th_2 type eytokines in EAN.Methods Lewis rats were grouped into EAN rats and control rats.The EAN rats were immunized by injection into both hind footpads of inoculums containing 100 ?g or 200 ?g of P2_(57-81)peptide and FCA while the control rats were immunized with FCA only.Clinical scores were compared at the maximum of disease.Supernatant productions of IFN- ?, IL-4 and IL-10 secreted by lymphocytes and obtained on day 14 after the immunization were examined. Histopathological assessment of sciatic nerves was made.Results Peak clinical scores of P2_(57-81)200 ?g (3.6?0.3)group were significantly higher than P2_(57-81)100 ?g group(2.2?0.6,P

The definition of critical quality attributes of Chinese materia medica ( CMM) was put forward based on the top-level design concept. Nowadays, coupled with the development of rapid analytical science, rapid assessment of critical quality attributes of CMM was firstly carried out, which was the secondary discipline branch of CMM. Taking near infrared (NIR) spectroscopy as an example, which is a rapid analytical technology in pharmaceutical process over the past decade, systematic review is the chemometric parameters in NIR model evaluation. According to the characteristics of complexity of CMM and trace components analysis, a multi-source information fusion strategy of NIR model was developed for assessment of critical quality attributes of CMM. The strategy has provided guideline for NIR reliable analysis in critical quality attributes of CMM.
Materia Medica ; standards ; Medicine, Chinese Traditional ; standards ; Spectroscopy, Near-Infrared ; methods

Objective To assess the proficiency of water monitoring laboratory at rural, county, and provincial levels in Guangdong province, to ensure the province's drinking water monitoring results accurate and reliable. Methods State quality of certified reference materials as water arsenic, fluoride and chloride of 90 copies each were numbered and distributed to 90 monitoring laboratories in the province for testing, The measurement results of the participatory labs were evaluated through normative values and expanded uncertainty, and were compared with those of robust statistics method. Results All participatory labs had timely feedback of their measurement results. The qualified rate was higher when arsenic was tested by hydride generation atomic fluorescence spectrometry and zinc-new silver salt of sulfuric acid spectrophotometric system, while fluoride and chloride by ion chromatography. The average qualified rates of water arsenic, fluoride and chloride of the province's rural drinking water quality monitoring laboratory were 66% (59/90), 72% (65/90) and 72% (65/90), respectively.Seven participatory labs failed the proficiency testing of all three analytes and unqualified rate was 7.8% (7/90)among the ninety participated monitoring labs. The qualified rates of robust statistics method for arsenic fluoride and chloride were greater than those evaluated by the expanded uncertainty, and large deviations with small sample sizes. Conclusions The testing ability of drink-water monitoring labs in Guangdong province has improved.However, by comparison with the requirements of national quality control and testing skills, there is still a gap. It is suggested that internal quality control be included in routine inspeetion to improve laboratory testing technology.

OBJECTIVETo construct a recombinant plasmid pIRES-GM-CSF-IL-21, and to investigate its antitumor effect on tumors in the mice.

METHODSFifty Bal b/c mice were included in this study. Cultured hepatoma H22 cells were inoculated in the left lobe of the liver to develop orthotopically transplanted liver tumor models. The mice with orthotopically transplanted liver tumor were randomly divided into 5 groups (n = 10): (1) Each mouse received injection of recombinant plasmid pIRES-GM-CSF-IL-21; (2) Received injection of plasmid pIRES-GM-CSF; (3) pIRES-IL-21; (4) Received injection of ampty plasmid pIRES (H22/neo group); (5) Received injection of PBS (H22 group) via the tail vein, respectively. Therefore, the anti-tumor effect was induced by both GM-CSF and IL-21, or by either of them alone. The serum levels of IFN-γ and IL-2 were detected by ELISA, and the cytotoxicity of spleen NK and CTL cells were tested by MTT colorimetry.

RESULTSComparing with the H22 and H22/Neo groups, the tumor weight in the mice of H22/GM-CSF group was (0.603 ± 0.223) g, H22/IL21-treated group (0.583 ± 0.290) g and H22/GM-CSF-IL21-treated group (0.303 ± 0.323) g, significantly lower than that in the H22 group [(1.591 ± 0.280) g] and H22/Neo group [(1.489 ± 0.155) g]. Among them the tumor growth was most significantly inhibited in the H22/GM-CSF-IL-21 group (0.303 ± 0.323) g, compared with that of H22 and H22/neo groups (P < 0.01). But there was no significant difference between the tumor weights of the H22/GM-CSF group and H22/IL-21 group, and between the tumor weights of the H22 and H22/Neo groups (P > 0.05). The levels of IFN-γ and IL-2 in peripheral blood of mouse models treated with H22/GM-CSF-IL-21 were significantly increased than that in the H22/GM-CSF group and H22/IL-21 group (all P < 0.01), but significantly decreased in the H22group and H22/Neo group (P < 0.01). The anti-tumor activity of splenic NK cells and CTLs in the H22/GM-CSF-IL21 group was significantly enhanced (P < 0.01), compared with the significantly decreased in the H22 and H22/Neo groups.

CONCLUSIONSOur results demonstrate apparent antitumor effect of the plasmid pIRES-GM-CSF-IL-21 on the orthotopically transplanted liver tumor in mice. The combination of both pIRES-GM-CSF and IL-21 is more effective than that of pIRES/IL21 or pIRES/GM-CSF treatment alone. In addition, the plasmid pIRES-GM-CSF-IL-21 can also promote the secretion of IFN-γ and IL-2 in vivo, and enhance the cytotoxic activity of splenic NK and CTLs against the transplanted liver tumor.

Animals ; Carcinoma, Hepatocellular ; blood ; pathology ; therapy ; Cell Line, Tumor ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Immunotherapy ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Interleukins ; genetics ; Killer Cells, Natural ; immunology ; Liver Neoplasms ; blood ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Plasmids ; therapeutic use ; Random Allocation ; Recombinant Proteins ; therapeutic use ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Burden

OBJECTIVETo compare clinical outcomes of swing shoulder and internal fixation in treating proximal humeral fractures.

METHODSFrom June 2007 to June 2012, totally 89 elderly patients with humeral proximal fractures were treated by swing of shoulder or internal fixation, and 81 patients were followed up. In swing shoulder group, there were 38 patients including 13 males and 25 females aged from 62 to 84 with an average of (67.11±6.18) years old; 27 cases were 2-part fractures and 11 cases were 3-part fractures according to Neer classfication. In internal fixation group, there were 43 patients including 16 males and 27 females aged from 60 to 80 with an average of (66.47±5.48) years old; and 29 cases were 2-part fractures and 14 cases were 3-part fractures according to Neer classfication. VAS score and complications were compared between two groups after treatment, and Constant-Murley functional scoring was used to evaluate shoulder function of patients.

RESULTSEighty-one patients were followed up from 13 to 26 months with an average of 18.3 months. There was no significant difference in preoperative VAS score between two groups. After treatment, VAS score in swing shoulder group was (3.11±0.95), and (3.88±1.14) in internal fixation group, and had significant difference between two groups (t=-3.313,P<0.05). There was no significant difference in Constant-Murley scores between swing shoulder group (79.53±3.73) and internal fixation group (77.98±4.11) (t=1.768,P>0.05). Postoperative complications in swing shoulder group was 18.4%(7/38), 39.5%(17/43) in internal fixation group, and had significant differences between two groups (χ2=4.313,P<0.05).

CONCLUSIONSwing shoulder for the treatment of proximal humeral fractures in elderly has advantages of low cost, less complications and good recovery of joint function; while internal fixation has a good therapeutic effect but increased complications.

Aged ; Aged, 80 and over ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Humans ; Male ; Manipulation, Orthopedic ; adverse effects ; methods ; Middle Aged ; Shoulder Fractures ; therapy

AIMTo establish quality control requirements and methods for recombinant adeno-associated virus(rAAV) type 2/human blood coagulation factor IX (rAAV-2/hFIX).

METHODSIdentification of rAAV genome fragments, potential contaminants including wild type AAV(wtAAV) and helper virus, were detected by PCR. Purity of rAAV-2/hFIX was analyzed by cation-exchange HPLC and SDS-PAGE. Virus partical numbers were performed by dot blot assay. hFIX expression was demonstrated by ELISA and potency of hFIX was verified by APTT.

RESULTSIdentity of rAAV-2/hFIX was proved. Residues of wtAAV and helper virus were conformed to requirements. Purity of rAAV-2/hFIX were more than 98%. Partical numbers of rAAV-2/hFIX were more than 1.0 x 10(15) VG.L-1. hFIX expression was more than 20.0 micrograms.L-1. hFIX potency was verified by APTT following rAAV-2/hFIX injected to FIX gene knockout mice, potency results conformed to requirements.

CONCLUSIONThe methods and requirements had been established for quality control of rAAV-2/hFIX.

Animals ; Dependovirus ; genetics ; Factor IX ; biosynthesis ; genetics ; Female ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Genome, Viral ; Humans ; Male ; Mice ; Mice, Knockout ; Quality Control ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics

Autoimmune Diseases ; diagnosis ; etiology ; therapy ; Child, Preschool ; Encephalitis ; diagnosis ; etiology ; therapy ; Female ; Humans ; Receptors, N-Methyl-D-Aspartate ; immunology

In order to develop characteristic folk medicine resources in Jiangxi, a pharmacognostical study was systematically performed for four different origin plants of Sikuaiwa, the result of study provides the microscopic features of powder and tissue of the crude drug. The research provided reference for the identification of Sikuaiwa, as well as a theoretical basis for the further development and the formulation of quality standards.
Magnoliopsida ; anatomy & histology ; chemistry ; growth & development ; Medicine, Traditional ; Plants, Medicinal ; anatomy & histology ; chemistry ; growth & development

OBJECTIVETo investigate the clinical features, pathology and treatment of a rarely seen laryngeal lesion, the vocal fold bamboo nodes.

METHODSTwo patients with vocal fold bamboo nodes were retrospectively reviewed. The clinical features, pathology and treatment were presented.

RESULTSVideo laryngoscope examination showed bilaterally creamy yellow transverse band like deposits in the submucosa, which were the typical vocal fold bamboo nodes in patients with autoimmune disease. Immunofluorescence pathology showed IgA, C1q and IgM deposition in lamina propria. Both patients were initially treated with oral hormones and one patient subsequently underwent submucosal resection of the lesion. The results of pathological and immunofluorescence investigations were reported in this paper, together with a discussion of the relevant literature.

CONCLUSIONSVocal fold bamboo node is a special laryngeal lesion in patients with autoimmune disease. The pathological results showed immune complexes in the vocal fold lamina propria. A surgical intervention can be applied if steroid drugs are not effective or the dysraphism of the vocal folds exists due to the bamboo nodes.

Adult ; Autoimmune Diseases ; pathology ; Female ; Humans ; Laryngeal Diseases ; diagnosis ; pathology ; therapy ; Laryngoscopy ; Retrospective Studies ; Vocal Cords ; pathology ; Young Adult