BACKGROUND:Accumulative evidence supports that vitreous cryopreservation can improve the cel survival rate. OBJECTIVE:To investigate the effect of vitreous cryopreservation on the tenocytes co-cultured with the porous polydimethylsiloxane (PDMS) scaffold. METHODS:Tenocytes were co-cultured with the porous PDMS scaffold for 9-14 days, and then preserved and resuscitated in the 10%dimethyl sulfoxide (DMSO), 21%DMSO and VS55, respectively. One hour later, the survival rate of post-resuscitated tenocytes versus pre-resusciated tenocytes was analyzed by live/dead double color fluorescent staining and flow cytometry. RESULTS AND CONCLUSION:Live/dead double color fluorescent staining revealed that tenocytes in the 10%DMSO group appeared to be irregular and double stained, and a large number of cel s shedding from the scaffold. The VS55 and 21%DMSO groups showed some spindle and hemispherical cel s single stained for green fluorescence and few double stained irregular cel s. Additional y, the cel density in the two groups was significantly lower than that in the control group. Flow cytometry results found that there were homogenous cel s in the control group;the number of cel s in the 10%DMSO group was too low to undergo flow cytometry;smal cel particles were visible in the VS55 group;in the 21%DMSO group, the cel volume was similar with the control group, and smal particles also existed. The survival rate in the VS55 group (64.9%) was significantly lower than that in the 21%DMSO group (76.2%;P<0.05). Conversely, the survived cel s were rare in the 10%DMSO group. To conclude, 21%DMSO vitreous cryopreservation improves the cel survival rate and is beneficial for tenocyte adherence to the scaffold.

Objective To investigate the clinical characteristics, diagnosis and treatment of acute mesenteric venous thrombosis(MVT) in the elderly. Methods The clinical features, diagnosis, treatments and prognosis of 10 aged cases with acute MVT were retrospectively analyzed. Results The chief complaints of the 10 cases were different degrees of abdominal pain, which not paralleled with abdominal signs. The accompanying symptoms were nausea, vomiting and bloody stools and so on. All of these patients were misdiagnosised as pancreatitis, appendicitis or intestinal obstruction and so on. diagnosis of two cases was confirmed by ultrasound, 8 by CT. At the same time, 2 cases underwent angiography examination. Of the 8 cases who underwent operation, 5 cases were cured, 3 cases died (1 died of toxic shock and 2 died of multiple organ failure ). Two cases underwent conservative intervention thrombolysis. Conclusions It is essential to improve the knowledge of acute MVT,especially its intricate clinical characteristics, high rates of misdiagnosis and mortality. Early proper diagnosis is crucial. The main treatment is operation and conservative intervention thrombolysis can be performed in the patients whose bowel has not necrosed yet.

OBJECTIVETo evaluate and compare the efficiency and safety of laparoscopic surgery (LS) and open surgery (OS) in the treatment of colorectal carcinoma.

METHODSRandomized controlled trials on laparoscopic surgery and open surgery for colorectal carcinoma from January 2000 to October 2010 were searched in the databases of EMbase, PubMed, Cochrane Library, Sciencedirect, Springer, VIP, CNKI, CBMdisc. The methodological quality was assessed according to the standard of Cochrane systematic review. For homogeneous studies, RevMan5.0 software was used for meta-analysis.

RESULTSA total of 13 RCTs involving 4603 patients were included in this study, and among those 6 were multi-center randomized controlled trials. The meta-analysis showed that: the operation time of the LS group was longer than that of the OS group (WMD = 38.91, 95%CI: 33.89 - 43.93, P < 0.001), the blood loss (WMD = -138.14, 95%CI: -195.79 - -80.50, P < 0.001) and the length of hospital stay (WMD = 2.91, 95%CI: -4.65 - -1.17, P = 0.001) of the LS group was less than those in OS group. There was no significant differences between the two groups in the number of dissected lymph nodes (WMD = -0.62, 95%CI: -1.47 - 0.23, P = 0.150). There was no significant differences between the two groups in terms of the postoperative complications (30 days) (RR = 0.78, 95%CI: 0.59 - 1.01, P = 0.06). There was no significant differences between the two groups in 3-year overall survival (RR = 1.00, 95%CI: 0.96 - 1.04, P = 0.970). There was no significant differences between the two groups in 5-year overall survival (RR = 1.03, 95%CI: 0.99 - 1.08, P = 0.140). There was no significant differences between the two groups in 5-year overall recurrence (RR = 0.89, 95%CI: 0.74 - 1.07, P = 0.200).

CONCLUSIONSLaparoscopic surgery for colorectal carcinoma is a safe and effective therapy as open surgery in the short term or long term outcomes. It could be an acceptable alternative to open surgery for colorectal carcinoma.

Colorectal Neoplasms ; surgery ; Humans ; Laparoscopy ; Randomized Controlled Trials as Topic ; Treatment Outcome

OBJECTIVETo research the constituents in needles of Pinus sylvestris.

METHODRepeated column chromatography and preparation HPLC are used for compound isolation, and their structures were elucidated on the basis of spectral data analysis.

RESULTSix compounds, pinifolic acid (1), 15-oxo-8 (17) -labden-18-oic acid (2) , 15-acetoxy-labd-8 ( 17)-en-18-oic acid (3), dehydroabietic acid (4), 7alpha-hydroxydehydroabietic acid (5), 7beta-hydroxydehydroabietic acid (6) were isolated from the needles of P. sylvestris.

CONCLUSIONCompound 3-6 were isolated from the needles of P. sylvestris for the first time, and compound 3 is a new natural product. The petroleum ether and EtOAc extracts showed significant cytotoxic effects to Hela and A549. Compounds 2, 4-6 revealed a positive distinction compared to the control group.

Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Chromatography, High Pressure Liquid ; Diterpenes ; chemistry ; isolation & purification ; pharmacology ; Diterpenes, Abietane ; chemistry ; isolation & purification ; pharmacology ; HeLa Cells ; Humans ; Molecular Structure ; Pinus sylvestris ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Structure-Activity Relationship

OBJECTIVETo study the chemical constituents of Potentilla chinesis and their anticancer activities.

METHODChemical constituents were isolated by repeated column chromatography (Toyopearl HW-40C and preparative HPLC). The structures were elucidated on the basis of spectral data analysis. The isolated compounds were screened with two anticancer models.

RESULTFifteen triterpenes, alpha-amyrin (1) , beta-amyrin (2) , ursolic acid (3) , corosolic acid (4), euscaphic acid (5) , pomolic acid (6) , tormentic acid (7), 2alpha, 3alpha-dihydroxyurs-12-en-28-oic acid (8), 2beta, 3beta, 19alpha-trihydroxyurs-12-en-28-oic acid (9), asiatic acid (10) , 24-hydroxy tormentic acid (11) , myrianthic acid (12), oleanolic acid (13), maslinic acid (14) and 2alpha, 3alpha-dihydroxyolean-12-en-28-oic acid (15) , were isolated from P. chinesis.

CONCLUSIONCompounds 1, 2, 4 -15 were isolated from the plant for the first time. Compounds 4, 8 - 10, 12, 14 and 15 show anticancer activities. Compounds 4, 9 show strong activities.

Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line ; Cell Survival ; drug effects ; Chromatography, High Pressure Liquid ; Fibroblasts ; cytology ; drug effects ; HeLa Cells ; Humans ; Mice ; Molecular Structure ; Plants, Medicinal ; chemistry ; Potentilla ; chemistry ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

In order to establish the stable andreliable ISSR-PCR System of Lysimachia christinae, L16 (4(5)) orthogonal design, which based on 7 levels of single factor experiment, were used in this study. The variance analysis was carried out by SPSS 19.0, and 5 main factors affecting the reaction system were optimized in 4 levels. The best annealing temperature was selected by the optimized reaction system. And the stability and reliability of this system was tested by 23 samples from different origins. The results showed that the five factors (DNA template, primer, dNTP, Mg2+ and Taq enzyme) were the most impacts on the amplified results of ISSR-PCR of L. christinae. The order of the influence was: primer > Taq enzyme > DNA template > Mg2+ > dNTP. The optimal system, which was determined by multiple comparison on different levels of each factor, was total volume of 25 microL, including DNA template 60 ng, primer 0.3 micromol x L(-1), dNTP 0.2 mmol x L(-1), Mg2+ 1.8 mmol x L(-1), Taq enzyme 1.25 U. The optimal system was stable and reliable tested by 23 samples from different origins. This study lays the foundation for genetic diversity analysis, fine varieties selection and molecular identification of L. christinae, and provides reference for optimization on ISSR-PCR system of other speciesin future.
DNA Primers ; genetics ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Primulaceae ; classification ; genetics ; Quality Control

Objective: To investigate the incidence of CMV infection(CMV-I) and CMV related diseases (CMV-D) after allogeneic hematopoietic stem cells transplantation in 70 consecutive allogeneic hematopoietic stem cells transplantation(allo-HSCT) patients and to search for the optimal prophylactic strategy.Methods: Blood samples were monitored using the CMV pp65 antigenemia assay.Of the 70 patients observed,30 patients with chronic myeloid leukemia［CML:CP(27),AP(2),BC(1)］,12 with acute myeloblastic leukemia(AML),10 with acute lymphoblastic leukemia(ALL)and other cases were NHL(3), AA(5), MDS(7), SCLC with pancytopenia (1),CLL(1), and MF (1). Sixty six patients received HLA - identical siblings transplantation and four received tranplants from their HLA- haploidentical donors. Seventy cases included allo-PBPCT (64 cases) , allo-BMT (4 cases) and allo-PB+BMT (2). Before transplantation, all patients and donors received CMV serological examination except 4 pairs of donors/recepients. All 66 patients (3 cases were CMV IgM positive) and 64/66 donors were CMV IgG positive. Results：After transplantation, 64/70 patients developed CMV viremia during monitoring period. Forty three of 70 patients developed CMV-D.Thirty five of them suffered from CMV-associated interstitial pneumonia(CMV-IP). The high peak levels of CMV antigenemia were associated with development of CMV disease　. Close correlation was found between acute graft vs host disease(GVHD) and CMV disease. The patients were followed up for 2 to 24 months. The patients who received preemptive therapy(group A)had significantly better outcome than CMV disease group(group B, P=0.0001). Conclusions: The results suggest that CMV antigenemia has high predictive value for subsequent CMV disease and CMV pp65 antigenemia -guided early therapy has particular advantage for avoiding morbidity and mortality caused by CMV disease.

OBJECTIVETo classify the Chinese isolates of Coltiviruses.

METHODSThree sets of primers were selected among them two were specific to the 9th and 12th segments of subgroup B2, and one was for the 12th segment of subgroup B1-All the Chinese isolates of Coltivirus selected in the experiment were classified according to the lengths of different amplicons of the reverse transcriptase-polymerase Chain reaction (RT-PCR). The homogenicity of the nucleic acids of the isolates BJ95-75 and YN-6 was also compared with other Coltivirus strains belonging to subgroup B2.

RESULTSWith the primers 12-854-S/12-B2-R, which were specific to the 12th segment of Coltivirus subgroup B2-850 bp amplicons were obtained from Beijing isolate BJ95-75 and all the Yunnan isolates such as YN-6, -67-1, -68-1, -69, -70-1, -70-2, -90, -92-2, -93 of Coltivirus 492 bp DNA fragments were also amplified from all of them with the segment 9th specific primers 9-JKT-S/9-JKT-R. However no positive results were obtained from Northeast isolates NE97-12, NE97-31 and control viruses YN-99(Orbivirus),YN-151-1(JEV) with the same two sets of primers. With 12-B1-S/12-B1R primers specific to the 12th segment of subgroup B1, no amplicons of right length were obtained from any of the Chinese isolates of Coltivirus and the control viruses. When compared the nucleic acid sequences of BJ95-75 and YN-6 with other Coltivirus strains such as Bannavirus, JKT6423, JKT6969, JKT7043, the amplicons from segment 12th of these two strains had more than 89.4% homology with the other strains, especially to the earlier Chinese isolate Bannavirus, the homolog was more then 98.9%. Nearly 96.5% and 99.2% of the nucleic acids of the amplicons from segment 9th of the two strains were being homologous to Bannavirus and about 84.0% to JKT6423, which had been classified into type B2a. But the maximal homogenicity was about 53% when compared with the other two coltivirus strains. JKT6969 and JKT7043 which had been classified into type B2b.

CONCLUSIONGenotyping the recent Chinese isolates of coltivirus for the first time in our country. Most of the Chinese isolates belong to subgroup B2, more exactly type B2a. The Northeast isolates NE97-12 and NE97-31 were not correctly grouped with the available primers.

Animals ; Base Sequence ; China ; Coltivirus ; classification ; genetics ; isolation & purification ; Culicidae ; virology ; Genotype ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Nucleic Acid

Objective To explore the significance of urine deoxypyridinoline to diagnosis on osseous metastasis of lung neoplasms.Methods.182 cases with lung carcinoma was divided into two groups.One group was case with osseous metastasis,the other group was case without osseous metastasis,uDPD/uCr, uCa/Cr,sCa and sAKP in two groups were respectively compared.Sensitivity and specificity of these indexes to diagnosis on osseous metastasis of lung cancer were also acalculated and compared.80 cases without osseous metastasis were follow-up for 6 months.Results The ratio of uDPD/uCr with osseous metastasis group[(12.35?2.65)nmol/mmol]was significantly higher than that of without osseous metastasis group [(7.76?2.11)nmol/mmol](t=2.46,P