Objective To assess the value of central venous-to-arterial carbon dioxide difference [ P( cv-a) CO2 ] in evaluation of disease severity and prognosis in patients with septic shock.Methods There were 45 consecutive resuscitated septic shock patients from April 2009 to October 2010 included immediately after their admission into our ICU.The patients were divided into low P(cv-a) CO2 group and high P(cv-a) CO2 group according to a threshold of 6 mm Hg ( 1 mm Hg=0.133 kPa).All patients were treated by early goal directed therapy (EGDT).The parameters of hemodynamics,lactate clearance rate,the Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ ) score,the sequential organ failure assessment (SOFA)score,6 h rate of EGDT achievement,the ICU mortality and 28 days in-hospital mortality were recorded for all patients.Results There were 30 patients in the low P(cv-a) CO2 group,and 15 in the high P(cv-a)CO2group.There were no significant differences between low P(cv-a) CO2 and high P(cv-a) CO2 patients in age,APACHE Ⅱ score and SOFA score (all P ＞ 0.05 ).Compared with the high P(cv-a)CO2 group,the low P(cv-a) CO2 group had higher cardiac index ( CI ) and 24 h CI,higher delivery O2 ( DO2 ) and 24 h DO2,higher central venous oxygen saturation ( ScvO2 ) [ (74 ± 9) ％ vs (67 ± 8) ％ ],lower lactate [ ( 3.4 ± 2.1 )mmol/L vs (5.7 ± 4.5 ) mmol/L] and higher △SOFA score [ (0.7 ± 1.8 ) vs ( - 0.4 ± 1.1 ) ],lower 24 h SOFA score [ (7.8 ± 2.0) vs (9.8 ± 2.0 ) ],higher 6 h rate of EGDT achievement ( 83.3％ vs 53.3 ％ )(P ＜ 0.05 ),however,there were no differences in 28 days mortality and ICU mortality between the two groups ( P ＞ 0.05 ).Conclusion P(cv-a) CO2 might be an indicator for predicting the severity of patients with septic shock and evaluating tissue perfusion.

Objective To express a recombinant protein TFPR1 ( the functional region of the snake venom proteins from Trimeresurus flavoviridis) in Pichia pastoris expression system. Methods The target gene was codon-optimized and synthesized according to the sequence of the conserved structural do-main of triflin and then cloned into the Pichia pastoris expression vector pPICZαA to construct the recombi-nant expression plasmid pPICZαA-TFPR1. The recombinant plasmid pPICZαA-TFPR1 was electroporated into the yeast strain X33. The transformed strains carrying expression plasmid were screened out with Zeocin and then induced by methanol to express the recombinant protein TFPR1. ELISA was performed for the screening of positive clones. SDS-PAGE and Western blot were used for further identification of the ex-pressed products. Results The recombinant plasmid pPICZαA-TFPR1 was successfully constructed. The recombinant protein TFPR1 was expressed in a secreted form at a molecular weight of 16×103. Conclusion The recombinant protein TFPR1 was successfully expressed in Pichia pastoris expression system, which laid a foundation for further researches on its biological function and application as an adjuvant.

OBJECTIVE: To determine the biological activity of ellagic acid extracted from gallnut against nasopharyngeal carcinoma and its molecular mechanism. METHODS: Nasopharyngeal carcinoma 5-8F cells were treated with 2, 4, 6 μg/mL ellagic acid for 48 h in vitro. The cell proliferation and cell apoptosis were analyzed by MTT and Hoechst33258 stain. The cell cycle and protein expression were measured by flow cytometry and Western blot. RESULTS: Ellagic acid inhibited the proliferation of 5-8F cells. The inhibition rates were (29.35±4.95)%, (53.32 ±4.44)% and ( 61.75 + 6.93)%, respectively, with significant difference from the control group (P<0.01). S phase cells in the experimental groups were (25.47±0.74)%, (28.08±1.41)% and (35.49±0.66)%, respectively, with significant difference (P<0.01) from the control group (21.26±0.70)%. Cells in the experimental groups showed nuclear pyknosis, karyorrhexis and poptotic cell morphology. The expression of COX-2 and stathmin in 5-8F cells was down-regulated with increased drug concentration. CONCLUSION Ellagic acid extracted from gallnut has activity against nasopharyngeal carcinoma cells, and its mechanism may be related to down-regulated expression of COX-2 and stathmin.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; genetics ; metabolism ; Down-Regulation ; drug effects ; Ellagic Acid ; pharmacology ; Humans ; Nasopharyngeal Neoplasms ; pathology ; Plant Extracts ; pharmacology ; Stathmin ; genetics ; metabolism

OBJECTIVETo study clinical characteristics of refractory or relapsed DNMT3A⁺ cytogenetically normal acute myeloid leukemia（CN-AML）patients, and to explore the overall response rate（ORR）and side effects of these patients followed the therapy including decitabine with CAG or CAGlike regimen.

METHODSIn this study we retrospectively analyzed 53 refractory or relapsed CN- AML patients receiving the therapy including decitabine combined with CAG and CAG- like regimen in our center from April 2011 to October 2014. The clinical characteristics and ORR were further analyzed. Based on gene mutations, these patients could be divided into 2 groups: DNMT3A⁺ AML patients（n=24）and DNMT3A- AML patients（n=29）.

RESULTSThe median age of DNMT3A⁺AML patients was 46 years old, higher white blood cells and bone marrow blasts were observed in DNMT3A+ AML group. The ORR and complete response（CR）rate of DNMT3A+ group were 62.50% and 54.17%, respectively. No differences were observed in ORR and CR rates（P>0.05）between these two groups. DNMT3A⁺/FLT3-ITD⁺ CN-AML patients（n=14）had higher ORR and CR rates than DNMT3A-/FLT3-ITD⁺CN- AML patients（n=15）（P= 0.040 and 0.042, respectively）. The one- year overall survival （OS） of DNMT3A⁺ AML group and DNMT3A- AML group were 59.58% , 54.09% , no differences were observed （P=0.438）. 25 patients received further therapy of allo-HSCT, the one-year OS of DNMT3A⁺ CN-AML was 87.50% and one-year disease free survival（DFS）was 72.73%, while the one- year OS was 61.54% and one- year DFS was 58.02% in DNMT3A⁻ group. No differences were observed between 2 groups （P=0.456, 0.217）.

CONCLUSIONDecitabine combined with CAG or CAG-like regimen was an effective and safe treatment for refractory or relapsed CN- AML patients. Compared to DNMT3A⁻/FLT3- ITD⁺ CN- AML patients, DNMT3A⁺/ FLT3-ITD⁺ CN-AML patients had higher ORR and CR rates. Decitabine bridged hematopoietic stem cells transplant could likely improve the survival of refractory or relapsed CN-AML patients.

Aclarubicin ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Azacitidine ; analogs & derivatives ; therapeutic use ; Cytarabine ; therapeutic use ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; Disease-Free Survival ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Leukemia, Myeloid, Acute ; genetics ; therapy ; Mutation ; Remission Induction ; Retrospective Studies

OBJECTIVETo explore the clinical and laboratory features of chronic myeloid leukemia (CML) with atypical e14a3 and e19a2 BCR-ABL fusion gene subtypes.

METHODSWe retrospectively analyzed a cohort of CML patients with Ph chromosome positive confirmed by cytogenetic and FISH but classical e13a3(b2a2), e14a2(b3a2)and e1a2 fusion transcripts negative identified by conventional real-time quantification RT-PCR (RQ-PCR). Further RQ-PCR was done with the forward primer and reverse primer designed to detect rare atypical BCR-ABL fusion genes including e14a3 and e19a2 transcripts. Direct sequencing analysis was performed on the PCR products and mutations in the BCR-ABL kinase domain were detected. The clinical data of patients were retrospectively analyzed.

RESULTSSix CML patients were found to carry t(9;22) abnormality and BCR-ABL rearrangement confirmed by FISH but classical BCR-ABL fusion genes negative detected by RQ-PCR. Further RQ-PCR and sequencing analysis confirmed the fusion of BCR exon 14 and ABL exon 3 in five CML patients (case 1-5) and the fusion of BCR exon 19 and ABL exon 2 in one CML patient (case 6). E255K and I293T IM-resistant mutations were detected in case 1 and 2, respectively. Among five cases with e14a3 transcripts, four were CML-CP, one CML-AP. Four patients were male and one was female. The median age was 48 years. The patient (case 6) with e19a2 transcripts was 40-year-old female with a diagnosis of CML-CP and PLT count was more than 1 000×10⁹/L. Imatinib (IM) therapy was administer in case 1, 2, 3, 4 and hematopoietic stem cell transplantation (HSCT) was undergone in case 5 after hydroxyurea (Hu) or interferon failure. Case 1 who had E255K IM resistant mutation, responded poorly to IM but obtained a complete cytogenetic remission (CCyR) after a substitution of dasatinib for IM. Case 2 and 3 achieved CCyR 6 months later after IM treatment and had been maintained well with IM despite I293T mutation in case 2. Case 4 attained CCyR 3 months later after IM treatment but relapsed and died soon. Case 5 was still in CCyR after HSCT. Case 6 with e19a2 transcripts got complete hematologic response after Hu treatment and CCyR was achieved soon after IM therapy.

CONCLUSIONIncidence of CML with atypical transcripts is extremely low. They could benefit from tyrosine kinase inhibitors or HSCT. Rare and atypical BCR- ABL fusion gene subtypes could be missed by conventional RQ-PCR.

Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; classification ; diagnosis ; genetics ; Male ; Middle Aged ; Retrospective Studies

Adolescent ; Adult ; Aged ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; therapy ; Male ; Middle Aged ; Mutation ; Pilot Projects ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; therapy ; Retrospective Studies

OBJECTIVETo investigate the epidemiology characteristics of norovirus among diarrheal outpatients in China.

METHODSDiarrhea cases were monitored at emergency/outpatient departments at 173 hospitals in 27 provinces of China, with clinical and epidemiological data, and fecal specimens collected and sent to 58 network-laboratories to detect norovirus by RT-PCR method, and to analyze the positive rate of norovirus in various regions, population and time during 2009-2013.

RESULTS11.6% of the 34 031 diarrheal cases under surveillance were found with norovirus. Age group of 6-23 month-old children and that of people over 45 years old were found with the highest positive percentage, 13.7% and 12.4% respectively. Positive percentage of norovirus peaks in autumn and winter in a year; it peaks in mid-temperate zones (10.7%) and warm-temperate zones (11.6%) in winter. It peaks in sub-tropical zones in autumn (14.3%). The most prevalent genogroups detected were norovirus G II, accounting for 89.9% of identified strains.

CONCLUSIONNorovirus affects all ages and was most prevalent in children and the elderly among diarrhea outpatients. Norovirus' positive percentage showed strong seasonal pattern, and peaks at different times of a year in different climate zones of China. Since no effective preventive measures existed, further study on norovirus epidemiology and intervention strategies should be conducted in future.

Adolescent ; Adult ; Aged ; Caliciviridae Infections ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Genotype ; Hospitals ; Humans ; Laboratories ; Middle Aged ; Norovirus ; Outpatients ; Prevalence ; Seasons