Objective To investigate the clinical efficacy of intraoperative methylprednisolone used in combination with perioperative enteral nutrition support in improving liver synthetic function after hepatectomy and postoperative recovery.Methods In this prospective controlled trial,79 patients treated with hepatectomy at Departement of Hepatobiliary Pancreatic Surgery in Drum Tower Hospital between June 2013 and May 2014 were included and divided into control group (n =39) and steroid group (n =40) according to the order of surgery.The control group underwent hepatectomy only,while the steroid group received intraoperative methylprednisolone (500 mg) at the start of liver excision.Both of the two groups were given the same perioperative enteral nutrition support.Liver function,prealbumin (PA),and C-reactive protein (CRP) levels were measured before and on days 1,3,and 5 after the operation.Human blood albumin infusion volume,postoperative gas passing,postoperative hospital stay,and incidence of complications were recorded.Results The serum level of PA on day 3 after hepatectomy was significantly higher in the steroid group than in the control group [(101.26 ±61.17) mg/L vs.(81.84 ±43.58) mg/L,t =-1.607,P =0.049].The serum level of cholinesterase on day 1 after hepatectomy was significantly higher in the steroid group than in the control group [(5.60±1.54) kU/Lvs.(4.68±1.01) kU/L,t=-3.136,P=0.004].On day1 and day 3 after hepatectomy,the serum levels of CRP were significantly lower in the steroid group than in the control group [(41.79 ±20.86) mg/L vs.(62.08 ±38.33) mg/L,t =2.933,P=0.027;(64.14 ±32.38) mg/L vs.(102.64 ± 49.05) mg/L,t =4.127,P =0.006].The postoperative hospital stay was significantly shorter in the steroid group than in the control group [(12.62 ±5.74) d vs.(15.41 ± 10.00)d,t =1.514,P =0.002].Conclusion Intraoperative use of methylprednisolone combined with perioperative enteral nutrition support may inhibit postoperative inflammatory response,promote early recovery of liver synthetic function,and help to promote rehabilitation after hepatectomy.

This study was purposed to construct three siRNA eukaryotic expression vector specific to mouse Qa-1 gene, to investigate its silencing effect on Qa-1 gene and to select the most efficient siRNA plasmid specific to mouse Qa-1 gene. Three siRNA peptides specific to mouse Qa-1 through siRNA Web design tools of Ambion company were chosed. Jingsai Company helped to complete the siRNA eukaryotic expression vector. The mouse NIH3T3 cells cultured in RPMI 1640 medium with 10% fetal bovine serum were divided into four groups: three groups of the cells were transfected with lipofectamine 2000 reagent and three different siRNA eukaryotic expression vectors, while one group cells were transfected with lipofectamine 2000 reagent alone as negative control. Cells were collected at 24, 48, 72 hours after transfection; the RNA level of Qa-1 was detected by RT-PCR, and the expression position was examined with flow cytometry analysis by using anti-Qa-1 monoclonal antibody. The results indicated that the constructed three siRNA eukaryotic expression vectors were found to be specific to mouse Qa-1 gene. The sequence analysis showed that the sequence was identical to what chosed from web tools. NIH3T3 cells in vitro were adhered in culture that cell shape appeared to change after transfection. RT-PCR and flow cytometry analysis by using anti-Qa-1 monoclonal antibody approved that both Qa-1 RNA and the expression of Qa-1 on cell surface decreased. The decreased levels in the three groups were different. At 24, 48 and 72 hours, the expression of Qa-1 on NIH3T3 cells decreased as in the following: H2-T231: 60.9%, 81.9%, 43.6%; H2-T232: 64.5%, 73.9%, 61.1%; H2-T233: 61.9%, 71.2%, 47.5%. H2-T232 was most efficient one in all three time points. It is concluded that all three siRNA eukaryotic expression vectors selected can successfully suppress the expression of the Qa-1, and from them H2-T232 is most efficient.
Animals ; Base Sequence ; Cells, Cultured ; Gene Silencing ; Genetic Vectors ; Histocompatibility Antigens Class I ; genetics ; Mice ; NIH 3T3 Cells ; Plasmids ; RNA, Small Interfering ; genetics ; Transfection

Gene Expression ; Graft Occlusion, Vascular ; prevention & control ; Humans ; Liver ; blood supply ; Stents ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate cyclooxygenase-2 (COX-2) expression and its relationship with mismatch repair (MMR) protein expression and microsatellite instability (MSI) in hereditary nonpolyposis colorectal cancer (HNPCC).

METHODSA total of 28 cases of colorectal adenoma and 14 cases of colorectal carcinoma were collected between July 2003 and July 2007 from 33 HNPCC families. Sporadic colorectal adenoma (n=32) and carcinoma patients (n=24) served as controls. With samples of tumor tissues and normal colonic mucosa collected from the patients, the protein expressions of COX-2 and MMR (hMLH1, hMSH2, and hMSH6) were examined with immunohistochemical assay. Frequency of MSI in five standard MSI loci BAT25, BAT26, D2S123, D5S346, and D17S250 were analyzed by means of polymerase chain reaction.

RESULTSThe rate of COX-2 high-expression was 53.6% (15/28) and 42.9% (6/14) in HNPCC adenoma and carcinoma; 62.5% (20/32) and 91.7% (22/24) in sporadic adenoma and carcinoma, respectively. That rate was lower in HNPCC carcinoma than in sporadic carcinoma (Pü0.05). MMR-deletion rate and percentage of high-frequency MSI (MSI-H) in HNPCC carcinoma were higher than those in sporadic colorectal carcinoma [both 71.4% (10/14) vs. 12.5% (3/24), both Pü0.01]. Among the 10 MMR-deficient HNPCC carcinoma patients, COX-2 low-expression was observed in 8 cases (80.0%), while COX-2 high-expression was observed in all of the 4 MMR-positive HNPCC carcinoma cases (Pü0.05). In comparison to MMR positive HNPCC carcinoma, HNPCC adenoma, and sporadic carcinoma, COX-2 expression was significantly lower in corresponding MMR-deficient cases (all Pü0.05). The rates of COX-2 low-expression in HNPCC adenoma, HNPCC carcinoma, and sporadic carcinoma with MSI-H were significantly higher than those in the cases with microsatellite stability (all Pü0.05).

CONCLUSIONCOX-2 is expressed at a low level in HNPCC carcinoma, different from the high COX-2 expression in sporadic carcinoma.

Adult ; Aged ; Base Pair Mismatch ; Base Sequence ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; Cyclooxygenase 2 ; genetics ; DNA Primers ; DNA Repair ; Female ; Humans ; Immunohistochemistry ; Male ; Microsatellite Repeats ; genetics ; Middle Aged

Background and Objective: Long-term use of cyclooxygenase-2 (COX-2) inhibitors can reduce the incidence of digestive cancers, such as colorectal cancers. Proliferating cell nuclear antigen(PCNA) is an important marker of cellular abnormal proliferation.This study was to evaluate the roles and correlation of COX-2 and PCNA in the onset and development of familial adenomatous polyposis(FAP) diseases.Methods: Thirty-six specimens of FAP adenomas tissues and 32 specimens of FAP carcinoma tissues from 11 FAP families, and 34 specimens of normal colonic mucosa were collected under colonoscopy from November 2004 to July 2007 in the General Hospital of Beijing Military Command.Immunohistochemistry was used to detect the expressions of COX-2 and PCNA. Results: The positive expression rates of COX-2 in normal colonic mucosa, FAP adenoma,and carcinoma tissues were 0(0/34),80.6%(29/36),and 93.8%(30/32),respectively.Proliferation index (PI) in normal mucosa,FAP adenoma,and carcinoma tissues were 17.79±7.49,34.47±10.57,and 71.75±9.22,respectively.Expressions of COX-2 and PCNA were significantly higher in the FAP adenoma and the carcinoma tissues than in the normal colonic mucosa (P＜0.01). The expression of PCNA was significantly higher in the FAP carcinoma tissues than in the FAP adenoma (P＜0.01). The expression of PCNA was higher in the FAP adenoma tissues with positive COX-2 than in the FAP adcnorma tussues with negative COX-2 (P＜0.01). Conclusions: COX-2 may play an important role in the development of FAP adenomas and colorectal carcinogenesis. COX-2 and PCNA may be important factors in the research on colorectal precancerous lesions and interventional therapy for colorectal neoplasm.

Objective To investigate the effect of laser artificial shrinkage(LAS)on pregnancy outcome in vitrification of human expanded blastocysts.Methods We selected 3859 frozen-thawed blastocyst-stage embryo transfers from January 2014 to December 2015.The transfers were divided into LAS group(n=3 176)and non-LAS group(n=683),which were then subdivided into ＜36 y subgroup and ≥36 y subgroup according to their age.Main outcomes measures were thawing rate,implantation rate and clinical pregnancy rate.Results Thawing rate, clinical pregnancy rate and implantation rate were 97.32%(5 453/5 603),66.81%(2 118/3 170),and 53.55%(2 912/5 438)in LAS group.In non-shrink group,they were 95.13%(1 173/1 233),62.70%(427/681),and 49.74%(582/1 170),which did not significantly differ from those in the former group(P＜0.05).Further analysis of the subgroups showed that thawing rate was significantly higher in LAS group than in non-shrink group of patients＜36 y(97.27% vs.95.33%;P＜0.05).Thawing rate and biochemical pregnancy rate were significantly higher in LAS group than in non-shrink group in patients ≥36 y(97.75% vs.93.66%;65.45% vs.50.65%,P＜0.05). Cancellation rate was not significantly different between the two groups(0.19% vs.0.29%, P ＞ 0.05). Conclusion LAS technique can increase thawing rate,clinical pregnancy rate and implantation rate before cryopreservation of blastocysts.

The purpose of this study was to investigate the regulatory effects of biofilm associated non-coding small RNA(sRNA)00085 on the survival and environmental fitness of Listeria monocytogenes.Homologous recombination technology was used to construct a deletion mutant strain(△sRNA 00085)and a complementary strain(C △sRNA 00085)of the sRNA00085 target gene.The differences in biological characteristics were compared among the standard strain,△sRNA 00085,and C△sRNA 00085.The deletion of sRNA00085 led to a significant decrease in biofilm formation capacity and sensitivity to several antibiotics,including penicillin,piperacillin,doxycycline,tetracycline,vancomycin,and cotrimoxazole.However,only the minimum inhibitory concentration(MIC)of tetracycline exhibited a significant decrease in △sRNA00085.Meanwhile,the decreased biofilm formation and antibiotic resistance of the sRNA00085 mutant were restored in the C△sRNA00085 strain.Furthermore,we investigated the transcription levels of tetracycline resistance-related genes in L.monocytogenes.Down-regu-lated transcription of the tetS gene but no significant difference in transcription of the tetA gene were observed in △sRNA 00085 compared with the standard strain and C△sRNA00085.Moreover,the elimination of sRNA00085 did not affect bacterial growth ability or sensitivity to disinfectants.These findings highlight that sRNA00085 plays an important role in the environ-mental adaptability of L.monocytogenes by affecting bacterial biofilm formation and resistance.

OBJECTIVETo study the feasibility of MRI of human colon adenocarcinoma cell line (Lovo) labeled with superparamagnetic iron oxide(SPIO) nanoparticles in vitro.

METHODSLovo cells (5 × 10(5) and 1 × 10(6)) were cultured in medium containing different SPIO nanoparticles (50 microl and 500 microl). Transmission electron microscopy was used to observe cellular ultrastructure and to determine the uptake and distribution of particles in Lovo cells at 1-, 3-, 6-hours. MRI of Lovo cells was performed with T1WI, T2WI sequences. Unlabeled cells were used as controls.

RESULTSUptake of SPIO nanoparticles occurred within 6 hours. On T1 weighted imaging, there was no significant difference in signal intensity between the experimental groups and the control group. On T2 weighted imaging, there was no significant difference in signal intensity between the experimental groups and the control group after culture of 1 h. Signal intensity began to decrease in 1 × 10(6) Lovo cells labeled with 500 microl SPIO nanoparticle after 3 hours culture. Signal intensity decreased in all the experimental groups after 6 hours culture.

CONCLUSIONHuman colon adenocarcinoma cell line (Lovo) can be labeled with SPIO nanoparticles, and the labeled cells can be imaged with MRI equipment.

Adenocarcinoma ; pathology ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Humans ; Iron ; Magnetic Resonance Imaging ; methods ; Magnetics ; Nanoparticles ; Oxides ; Pilot Projects ; Staining and Labeling ; methods

OBJECTIVETo evaluate the prevalence of TET2 gene mutation in acute myeloid leukemia (AML) patients, and analyze their clinical characteristics and prognosis.

METHODSPolymerase chain reaction (PCR) and direct sequencing were used to sequence exon 3 to 11 of TET2 gene.

RESULTSAmong 96 AML patients, TET2 gene mutation was detected in 13 (13.54%) patients (95%CI 6.70% - 20.38%). The median age was 54 years in mutated group and 41 years in unmutated group (P = 0.010). Mutated and unmutated patients did not significantly differ in gender, white blood cells (WBC) count at diagnosis, platelet count, PB and BM blast percentage and chromosome karyotype, excepting for hemoglobin level 84 (70 - 108) g/L in mutated group versus 70 (55 - 87) g/L in unmutated group (P = 0.032). TET2 gene mutation had no significant correlation with C-KIT, FLT3, JAK2V617F mutations, but did with NPM1 mutation. TET2 mutated patients had lower CR1 rate and 2-year overall survival than unmutated in non-M(3) patients (P < 0.05).

CONCLUSIONSTET2 gene mutation is more prevalent in older AML patients and has a certain correlation with clinical characteristics and outcome. It may be a molecular marker for poor prognosis in AML.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Exons ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Proto-Oncogene Proteins ; genetics ; Young Adult

OBJECTIVETo establish multiple short tandem repeat (STR) amplification by fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis for quantitative determination of chimerism, and to evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThirty-one patients received bone marrow transplantation (BMT) or nonmyeloablative allogeneic stem cell transplantation (NST) were evaluated. Peripheral blood and bone marrow were co-llected before and after transplantation in different period. Nine different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus PCR amplification kit. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype software were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient.

RESULTS48.4% of the patients received sex-matched transplantation and the quantification of donor chimerism could only be performed by STR-PCR method. Comparison of values obtained by FISH analysis with that by STR-PCR in patients transplanted from sex-mismatched donors showed an excellent correlation. The median number of informative alleles was 6.7 (range 2 - 10). The donor's alleles appeared in all the patients on day 7 post-transplant. The median values of donor chimerism in BMT group were inferior to that in NST group on day 7, day 14 and 1 month post-transplant. However the difference disappeared in the midterm or later period of transplant. On day 21, all of the 31 patients had stable engraftment and the percentage of donor chimerism was more than 92%. Median follow-up was 17 (3.5 - 29.0) months after transplantation. Twenty-six of 31 patients had durable engraftment and donor chimerism ratio was more than 90%. So for all of them survived leukemia-freely. Four of the 31 patients had unstable mixed chimerism and relapsed within 6 months post allo-HSCT. Another patient with unstable mixed chimerism appeared graft rejection. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and disease relapse. The incidence of GVHD was much higher in the group of full donor chimerism.

CONCLUSIONSequential and quantitative monitoring of STR is a valuable tool for studying engraftment dynamics, graft rejection, and relapse and for predicting GVHD. Furthermore it can provide a basis for early intervention of clinical treatment.

Adolescent ; Adult ; Child ; Electrophoresis, Capillary ; Female ; Graft Rejection ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Polymerase Chain Reaction ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous