Background Stickler syndrome is a genetic connective tissue disorder that affects the ocular,skeletal,orofacial and auditory systems.To determine the gene mutation loci can offer a basis for genetic diagnosis and management of Stickler syndrome.Objective The aim of this study was to research the clinical characteristics of a pedigree with Stickler syndrome and identify the disease-causing gene mutation.Methods This study was approved by Ethic Committee of Peking Union Medical College Hospital.The clinical study and pedigree analysis were performed in one family with Stickler syndrome type Ⅰ (STL Ⅰ).Nine family members were examined with informed consent.The entire coding regions of COL2A1 gene with flanking intronic regions were amplified by PCR and directly sequenced.The detected sequence change was confirmed to be mutationloci by examining whether they existed in normal control individuals.Mutant proteins were predicted with online software.Results There were 4 generations and 11 members in this family,and 2 members died,including 1 patient.Three patients were found in 9living families.Inheritance of this family complicd with an autosomal dominant inheritance mode.All affected individuals showed the consistent phenotypes with STL Ⅰ,including high myopia,membranous vitreous anomaly and surface central flat,short nose,palatoschisis,etc.Mutation screening of COL2A1 gene revealed that the first base of intron 12 was deleted(IVS12+1G del).Nucleotide sequence analysis showed that this mutation led to the functional abnormal of this gene by forming termination cordon in advance.This mutation occurred in all affected individuals,however,no mutation was observed in any unaffected member or 100 normal unrelated individuals.Conclusions This study identifies a novel splice-site mutation(IVS12+ 1G del)in COL2A1 gene in a Chinese STL Ⅰ pedigree.This is the first report on a mutation in a Chinese STL Ⅰ family.

To identify the structure of three related substances in potassium sodium dehydroandrographolide succinate (PSDS), an HPLC preparation method was used to separate the impurities. These main impurities were identified using LC-ESI/TOFMS, LC-ESI/MSn, NMR, UV and IR. One of the main impurities was a hydrolyzed and oxidized product of PSDS, which has not been reported previouely. The other two impurities were hydrolyzed products of PSDS after losing different succinic acids. The results indicate that PSDS can be easily hydrolyzed and oxidized. It should be stored at cool and dry places.

Aim To develop the in vitro and in vivo models induced by shrimp tropomyosin( ST) and mono-clonal tropomyosin-specific murine IgE antibody ( anti-ST-IgE mAb) . Methods ST was purified from Metap-enaeusensis by an isoelectric precipitation method. The anti-ST-IgE mAb was obtained from hybridomas. After RBL-2 H3 cells were sensitized with anti-ST-IgE mAb and challenged with ST,β-hexosaminidase release was determined. Passive systemic anaphylaxis ( PSA ) was induced in mice and the rectal temperature was recor-ded after ST challenge within 30 min by a thermal probe. Results A significant increase ofβ-hexosamin-idase was observed in sensitized cells after ST chal-lenge. The average temperature drop after ST challenge was 1. 44℃ in PSA mice within 30 min. Conclusion The in vitro and in vivo models induced by ST and anti-ST-IgE mAb are established as an improvement of pres-ent models of type Ⅰ allergy.

Objective:To develop murine models of Th2 response induced by shrimp tropomyosin (ST). Methods:Mice were sensitized with ST for 6 weeks. The serum antigen-special IgE (sIgE),total IgE and sIgG level,Th1/Th2 cytokines production were measured by ELISA. The basophil activation in mice was measured by flow cytometry. Results:The intraperitoneal sensitization with ST for 6 weeks induced significant increase of serum sIgE,total IgE and sIgG (sIgG1,sIgG2a and sIgG2b) level in mice. Th2 cell response was induced and cytokines (IL-4,IL-5,IL-10 and IL-13) production increased in splenocytes stimulated by ST,while Th1 cytokine (IFN-γ) production decreased. As the markers of basophil activation,CD200R and CD41 expression also increased in response to ST. Conclusion:The Th2 response is dominant in ST-induced anaphylaxis in mice.

Objective: To assess the effect of coronary collateral circulation (CCC) on myocardial viabilityin patients with chronic total occlusion of left anterior descending (LAD) artery. Methods: A total of 101 consecutive patients with confirmed diagnosis of total LAD occlusion in our hospital were enrolled. Rest 99mTc-MIBISPECT myocardial perfusion and 18F-FDG PET were performed, in addition all patients received coronary angiography (CAG) at 3 months front and back. Both images were reconstructed in the same machine and QPS software was used to obtain the summed rest score (SRS), abnormal resting total perfusion defect (TPD), viable and non-viable myocardium, LVEDV, LVESV and LVEF in relevant patients. Based on CAG result, the patients were divided into 2 groups: CCC group, n=39 and No CCC group, n=62; according to existing old myocardial infarctionand location of LAD occlusion, the patients were further divided into 4 subgroups. The above parameters were compared among different groups. Results: There were 86 male and 15 female patients with the mean age at (59.92±11.43) years. Relevant parameters in CCC group and No CCC groupwere as in SRS: (21.23±9.68) vs (28.56±8.76), TPD: (30.03±13.69) %vs (40.37±12.50) %, viable myocardium: (21.77±13.12) % vs (13.66±9.23) %, non-viable myocardium (8.28±8.58) %vs (27.40±12.97) %, all P<0.05; in LVEDV: (109.82±30.01) ml vs (173.71±57.69) ml, LVESV: (62.82±22.39) ml vs (122.53±51.66) ml, LVEF: (43.85±8.46) % vs (31.03±8.30) %, all P<0.05. Conclusion: Our preliminary study found that CCC could maintain left ventricular rest perfusion, myocardial viability and protect cardiac function in patients with chronic total LAD occlusion.

Objective To provide scientific evidence for the effectiveness of upper limb training after stroke.Methods Surface electromyograms (sEMGs) of the triceps braehii and biceps brachii were recorded in stroke patients during maximum isometric voluntary contraction (MIVC).A total of 18 patients with hemiparesis were studied.During the elbow's MIVC,flexor and extensor peak torque were measured,and sEMGs of the biceps and tri- ceps brachii were recorded.Results During MIVC,the biceps braehius of the intact side registered a stronger EMG than that of the affected side when the elbow flexed,but the differenees in the triceps braehii readings were not significant.The triceps bracbius of the intact side gave a stronger iEMG than the affected side when the elbow extend- ed,but the iEMG form the biceps brachius of the affeeted side was higher than that of the intact side.The co-contrac- tion ratio (CCR) of the triceps brachius on the affected side was higher than that of the intact side.Though there was a tendency for the CCR of the biceps brachius on the affected side to be higher than the intact side,any difference was not statistically significant.For both flexor and extensor MIVC,the peak torque on the affected side was lower than that of the intact side.Conclusions Elbow spastieity in hemiplegic patients is mainly attributable to the flexor muscles.In the rehabilitation of the upper limb after stroke,it is important when training extensor strength to inhibit co-contraction of the antagonistic muscle.

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

OBJECTIVE To reverse multidrug resistance(MDR)to chemotherapeutic agents in human laryngeal cancer cells(LSC-1/TAX)by knockdown the expression of P-glycoprotein(P-GP), the MDR1 gene product, using RNA interference(RNAi)technique.METHODS LSC-1/TAX cells were transfected with lentivirus vector that contains the shRNA construct targeting MDR1 mRNA.The drug resistance was measured by MTT assay in vitro, and sensitivity of the laryngeal cancer cells to various anti-neoplasm agents was quantified in vivo.The knockdown of MDR1 gene expression was assessed by immunocytochemistry in vitro and vivo.RESULTS Multidrug resistance phenotype in human laryngeal cancer cell line (LSC-1/TAX)was reversed in vitro.Tumor growth assay in vivo revealed a reverse of MDR in the laryngeal cancer cells.Immunocytochemistry showed that P-GP expression was significantly inhibited.CONCLUSION MDR1 shRNA lentivirus vectors can significantly inhibited MDR1 expression.Inhibition of MDR1 gene expression conferred an increased sensitivity of drug-resistant in laryngeal cancer cells to conventional chemotherapeutic agents.

OBJECTIVETo detect the expression of fibroblast activation protein(FAP) in colorectal cancer tissue, and to investigate the association between expression of FAP with pathological parameters.

METHODSFifty-five cancer tissues and 50 normal colorectal samples were examined using immunohistochemistry with anti-FAP polyclonal antibody. The distribution of positive cells in different tissues, and associations of positive cell number with tumor staging, lymph node metastasis and tumor invasion were investigated to evaluate the effects of FAP on pathological progress in colorectal cancer.

RESULTSNo FAP expression was observed in 50 normal colorectal tissue samples. FAP positive cells were seen in carcinoma associated fibroblasts(CAFs), and in few colorectal cancer cells. The numbers of FAP positive cells in tissue samples of TNM III(-IIII((40.1±15.9) was significantly greater than that of TNMI(-II( (18.3±7.7)(P<0.01). Furthermore, the number of FAP positive cells in tissue samples with lymph node metastasis (44.4±13.3) was also significantly higher than those without lymph node metastasis (18.5±8.1)(P<0.01). Significant positive correlations were found between the number of FAP-positive cells with the tumor TNM staging and lymph node metastasis(r=0.544 and r=0.793, respectively)(P<0.01). The number of FAP-positive cells was 25.2±8.9 in T2, 32.41±19.30 in T3, and 29.2±16.5 in T4. The association between number of positive cells and depth of invasion was not statistically significant(P>0.05).

CONCLUSIONSThe FAP mainly expresses in CAFs locating in colorectal cancer tissues. The number of FAP positive cells is positively correlated with TNM staging of colorectal cancer and lymph node metastasis.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Gelatinases ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Serine Endopeptidases ; metabolism

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment