Objective: To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on cell proliferation and apoptosis after transferred into SW1990 cell line. Methods: K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR. Clones with inverted insertion were selected and co-transferred into E. coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination. Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus. Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified; the virus titer was determined. Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining. SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay. Results: A 282 bp target gene fragment was acquired by PCR; the titer of recombinant adenovirus was 7.6 × 108 pfu/ml before purification by CsCl2 gradient centrifugation and 5.0 × 1910 pfu/ml after CsCl2 gradient centrifugation. When the recombinant adenovirus was at 100 MOI, the infection efficiency of SW1990 cells nearly reached 100%. The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation (P<0.05), with a maximal inhibitory rate of 40.5% 4-5 days after infection; it also significantly increased SW1990 cell apoptosis, with the apoptotic rate being (22.54 ± 5.38) % 72 hours after infection. Conclusion: We have successfully constructed an anti-sense RNA adenovirus vector targeting K-ras exon 1 of SW1990 cell line, paving a way for the anti-sense K-ras gene therapy of pancreatic carcinoma.

AIMTo study the chemical constituents of the stems and leaves of Aconitum coreanum (Lèvl.) Rapaics.

METHODSThe constituents of Aconitum coreanum were isolated by using various kinds of modern chromatographic methods. The new alkaloid was identified on the basis of spectral analysis.

RESULTSTwo compounds were isolated and identified as: 13-dehydro-1beta-acetyl-2alpha,6beta-dihydroxyhetisine (I) and Guanfu base G (II).

CONCLUSIONCompound I is a new alkaloid.

Aconitum ; chemistry ; Diterpenes ; Heterocyclic Compounds, Bridged-Ring ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the bacterial distribution and drug resistance in patients with surgical infections, and provide the basis for the standardization treatment of the surgical infection.

METHODSRetrospectively analyzed from January 2008 to December 2011 surgical infection in our samples bacteria identification and drug sensitivity test results.

RESULTSA total of 3829 nonduplicate isolates from 3257 samples, Gram-negative bacteria accounted for 62.4% (the main microbes were P.aeruginosa, K. pneumonia and E.coli etc) and Gram-positive bacteria accounted for 37.6% (the main microbes were Enterococcus, Staphylococcus and coagulase negative Staphylococcus). Incidence of Staphylococcus aureus and Enterococcus faecalis were on an obvious increase. For the performance of the high level of sensitive to Imipenem, Amikacin, Piperacillin and Tazobactam by E. coli and K. pneumonia. The Pseudomonas aeruginosa and Acinetobacter baumannii to cephalosporins, Carbapenems and Fluoroqinolones were higher resistant with Multidrug resistance. No vancomycin and teicoplanin resistant Enterococcus faecium were found. The prevalence of ESBL E.coli was 45.6%-61.5% and ESBL K.pneumoniae isolates were fluctuated. The methicillin-resistant S.aureus (MRSA) isolates were relatively high (21.1%-55.8%), and methicillin-resistant Staphylococcus epidermidis was higher than the other Gram-positive cocci. Vancomycin for Staphylococcus performance was highly sensitive.

CONCLUSIONSThe main composition of surgical clinical infection pathogens are Gram-negative bacillus, and the emergency of resistance of bacteria to antibacterial drugs is a common phenomenon. The resistant rate shows ascendant trend; Drug resistance is significantly higher in Pseudomonas aeruginosa and Acinetobacter baumannii. Antimicrobial resistance is a serious and challenging issue.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anti-Bacterial Agents ; pharmacology ; Bacteria ; drug effects ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Retrospective Studies ; Surgical Wound Infection ; microbiology ; Young Adult

BACKGROUNDBloodstream infections (BSIs) remain a major cause of morbidity and mortality in patients undergoing surgery. This study aimed at elucidating the clinical characteristics of community-acquired BSIs (CABs) and nosocomial BSIs (nBSIs) in patients admitted to the surgical wards of a teaching hospital in Beijing, China.

METHODSThis cross-sectional study compared 191 episodes of BSIs in 4074 patients admitted to the surgical wards between January 2008 and December 2011. Cases of BSIs were classified as CABs or nBSIs, and the characteristics, relevant treatments, and outcomes of CABs and nBSIs were compared.

RESULTSOf the 191 BSIs, 52 (27.2%) and 139 (72.8%) were CABs and nBSIs, respectively. Escherichia coli, coagulasenegative staphylococci, and Klebsiella spp, were the most frequently isolated microorganisms. There were significant differences between CABs and nBSIs with respect to the use of hormonal drugs, ventilation, acute physiology and chronic health evaluation (APACHE) II and American Society of Anesthesiologists scores, and prevalence of cancer (P < 0.05). Empirical antibacterial therapy did not decrease the crude mortality, but multivariate analysis showed that high APACHE II was independently associated with a risk of mortality (odds ratio = 0.97, 95% confidence interval: 0.93-1.02 for APACHE II).

CONCLUSIONSWe found significant differences in the clinical characteristics of surgical patients with CABs and nBSIs. The outcome of patients seems to be related to high APACHE II scores.

Anti-Bacterial Agents ; Bacteremia ; epidemiology ; China ; Community-Acquired Infections ; epidemiology ; microbiology ; Cross Infection ; epidemiology ; microbiology ; Cross-Sectional Studies ; Escherichia coli ; pathogenicity ; Female ; General Surgery ; statistics & numerical data ; Hospitals ; Humans ; Male ; Staphylococcus ; pathogenicity

OBJECTIVETo investigate the ultrastructural changes of duodenal mucosas and their significance in patients with liver cirrhosis (PLC).

METHODSEndoscopic biopsy duodenal mucosa specimens of 60 PLC and 18 healthy volunteers as controls were obtained. Ultrastructural changes of them were studied with transmission electron microscopy. These PLC were divided into groups A, B and C according to the Child-Pugh classification. The ultrastructural changes in the duodenal mucosas of each group were rated and compared with those of the other groups. PLC with and without ultrastructural changes of duodenal mucosas were divided into a positive group and a negative group. Levels of plasma Alb, TBil, PT, plasma endotoxin, and blood ammonia of the PLC were detected and compared.

RESULTSThere were 20 PLC each in groups A, B, and C. Ultrastructural changes of duodenal mucosas were found in 5 PLC of group A, 9 in group B and 17 in group C. Among the 60 PLC, 52% had some changes in their duodenal mucosas. The changes included decrease and rupture of the microvilli; also karyopyknosis, karyorrhexis, widening of the gaps of the tight junction and tumefactions of mitochodrion of duodenal mucosa epithelial cells. No ultrastructural changes of duodenal mucosas were found in the control group. The rate of changes in the three Child-Pugh class groups and in the control group were 25%, 45%, 85%, 0% respectively (P < 0.01). The level of Alb of the positive group was significantly lower than that of the negative group (P < 0.01). Levels of plasma TBil, PT, endotoxin and blood ammonia of the positive group were significantly higher or longer than those of the negative group (P < 0.01). Levels of plasma Alb of the positive and negative groups were significantly lower than those of the control group (P < 0.01). Levels of TBil, PT, plasma endotoxin and blood ammonia of the positive and negative groups were significantly higher or longer than those of the control group (P < 0.01).

CONCLUSIONThere were ultrastructural changes of duodenal mucosas in PLC, especially in end-stage PLC. Ultrastructural changes of intestinal mucosas in the PLC may have important pathophysiological and clinical significance.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Intestinal Mucosa ; pathology ; ultrastructure ; Intestine, Small ; pathology ; ultrastructure ; Liver Cirrhosis ; pathology ; Male ; Middle Aged

BACKGROUNDSince January 2002, adult-to-adult living donor liver transplantation (AALDLT) has gained increasing popularity in China in response to the shortage of cadaveric donor livers. This study presents a detailed analysis of the outcomes of AALDLT in a single center.

METHODSA total of 70 patients underwent AALDLT at our center between January 2002 and January 2007. Among these, 67 patients received a right lobe graft without the middle hepatic vein and 3 patients received dual grafts. Three-dimensional volumetric computed tomography, magnetic resonance imaging with angiography and cholangiography were performed preoperatively. Recipient operation time, intraoperative transfusion requirement, length of intensive care unit stay, length of hospital stay, liver function tests, coagulation tests and surgical outcomes were routinely investigated throughout this study.

RESULTSAll donors survived the procedure with an overall complication rate of 15.3%. Overall recipient 1-year survival and complication rates were 87.1% and 34.2%, respectively. Among the 70 cases, average graft recipient weight ratio was 0.94% (0.72% - 1.43%) and average graft volume/standard liver volume ratio was 46.42% (31.74% - 71.68%). All residual liver volumes exceeded 35%. Liver function and coagulation recovered rapidly within the first 7 days after transplantation.

CONCLUSIONSAALDLT is a safe procedure for the donors and an effective therapy for patients with end-stage liver disease. Patient selection and timely decision-making for transplantation are essential in achieving good outcomes. With accumulation of experience in surgery and clinical management, timely feedback and proper modification, we foresee better outcomes in the future.

Adolescent ; Adult ; Aged ; Female ; Humans ; Liver Transplantation ; methods ; Living Donors ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective: To investigate the association of the abnormal length of human Y chromosome with semen quality and the outcome of assisted reproductive technology (ART). METHODS: Based on the karyotype, we assigned the patients undergoing ART to a normal control, a long Y chromosome (Y>18), and a short Y chromosome group (Y<22). We compared the semen parameters and numbers of embryos and high-quality embryos among the three groups of patients and performed statistical analysis of the obtained data using Chi-square distribution and t-test. RESULTS: Compared with the control, the Y>18 group showed a significantly lower incidence rate of asthenozoospermia (31.03% vs 8.33%, P <0.05) and a larger number of high-quality embryos (5.46 ± 4.54 vs 7.40 ± 5.49, P<0.05). Both the incidence rate of azoospermia and number of total embryos were remarkably lower in the control than in the Y<22 group (1.87% vs 16.47%, P <0.05; 8.60 ± 7.03 vs 10.00 ± 6.58, P<0.05). No statistically significant differences were found in the pregnancy rate between the Y>18 and Y<22 groups (P>0.05). CONCLUSIONS Short Y chromosome may affect spermatogenesis, but the length of Y chromosome does not negatively influence the outcome of ART.
Asthenozoospermia ; genetics ; Azoospermia ; genetics ; Chi-Square Distribution ; Chromosomes, Human, Y ; Female ; Humans ; Karyotype ; Karyotyping ; Male ; Pregnancy ; Pregnancy Rate ; Reproductive Techniques, Assisted ; Semen ; Semen Analysis ; standards ; Sex Chromosome Aberrations ; Spermatogenesis ; Treatment Outcome

Acute myeloid leukemia (AML) is a malignant clonal disease of hematopoietic stem cells, characterized by the proliferation of abnormal primordial cells of myeloid origin in bone marrow, blood and other tissues. At present, the standard induction therapy for AML mainly includes “3+7” standard treatment(anthracycline combined with cytarabine), allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and targeted drug therapy. However, AML cells usually express high levels of P-glycoprotein, which mediates the efflux of chemotherapeutic drugs, which makes AML cells resistant to chemotherapy, resulting in many patients who are not sensitive to chemotherapy or relapse after complete remission. And some patients can not tolerate intensive therapy or lack of donors and can not use Allo-HSCT therapy. Therefore, it is of great clinical significance to find new drugs to improve the efficacy of AML patients. Epigenetic disorders play a key role in the pathogenesis of many diseases, especially cancer. Studies have shown that most AML patients have epigenetic regulatory gene mutations, such as DNMT3A, IDH and TET2, and these mutations are potentially reversible, which has become one of the therapeutic targets of AML. Histone deacetylase inhibitors (HDACi) can regulate the balance between histone acetylation and deacetylation, change the expression of proto-oncogenes or tumor suppressor genes that control cancer progression from epigenetics, and play an important role in many kinds of tumor therapy. At present, HDACi has shown the ability to induce differentiation, cell cycle arrest and apoptosis of AML cells. The mechanism may be mainly related to HDACi inducing chromatin conformation opening of tumor suppressor gene by inhibiting HDAC activity, promoting oncogene damage and preventing oncogene fusion protein from recruiting HDAC. Although the preclinical outcome of HDACi is promising, it is not as effective as the conventional therapy of AML. However, the combination strategy with various anticancer drugs is in clinical trials, showing significant anti-AML activity, improving efficacy through key targeting pathways in a typical synergistic or additive way, increasing AML sensitivity to chemotherapy, reducing tumor growth and metastasis potential, inhibiting cell mitotic activity, inducing cell apoptosis, regulating bone marrow microenvironment, which provides a good choice for the treatment of AML. Especially for those AML patients who are not suitable for intensive therapy and drug resistance to chemotherapy. This review introduces the relationship between HDAC and cancer; the classification of HDAC and its function in AML; the correlation between HDAC and AML; the clinical application of five types of HDACi; preclinical research results and clinical application progress of six kinds of HDACi in AML, such as Vrinota, Belinostat, Panobinostat, Valproic acid, Entinostat, and Chidamide, the mechanism of HDACi combined with other anticancer drugs in AML indicates that the current HDACi is mainly aimed at various subtypes of pan-HDAC inhibitors, with obvious side effects, such as fatigue, thrombocytopenia, nausea, vomiting, diarrhea. In recent years, the next generation of HDACi is mainly focused on the selectivity of analogues or isomers. Finding the best combination of HDACi and other drugs and the best timing of administration to balance the efficacy and adverse reactions is a major challenge in the treatment of AML, and the continued development of selective HDACi with less side effects and more accurate location is the key point for the development of this drug in the future. It is expected to provide reference for clinical treatment of AML.

OBJECTIVETo evaluate the safety of meningococcal group AC bivalent polysaccharide conjugate vaccine among children aged 5-24 months old.

METHODSFrom July 2011 to June 2012, a total of 34 411 children aged 5-24 month-old who voluntarily vaccinated meningococcal group AC bivalent polysaccharide conjugate vaccine in Zhongshan city were included. The adverse effects within 72 hours were recorded and analyzed.

RESULTS34 411 children were recruited, including 18 708 boys (54.36%), whose mean age were ( 11.4 ± 3.9 ) months old.Within 72 hours, the incidence rates of local adverse effects were 0.76% (261/34 411) for erythema,0.57% (197/34 411) for sclerosis,0.56% (191/34 411) for swelling,0.42% (143/34 411) for pain,0.15% (53/34 411) for pruritus, and 0.15% (50/34 411) for rash on the injection site. The overall incidence rate of local adverse effects was 1.61% (554/34 411; 95%CI:1.48%-1.74%). The incidence rates of systemic adverse effects were 0.98% (312/34 411) for fever,0.48% (164/34 411) for anorexia,0.31% (108/34 411) for diarrhea,0.29% (100/34 411) for malaise,0.20% (70/34 411) for nausea and vomiting, and 0.08% (26/34 411) for headache. The overall incidence rate of systemic adverse effects was 1.64% (565/34 411; 95%CI:1.51%-1.78%).25 children (0.07%) had hyperpyrexia ( > 39°C), and the time of duration lasted less than 48 hours.16 children (0.05%) had symptoms of cold, such as cough and catarrh.No accident and other serious events were reported. The incidence rate of systemic adverse effects among boys was 1.79% (334/18 708), which was higher than that of girls (1.47%, 231/15 703), the difference showed statistical significance (χ(2) = 5.22, P < 0.01). The incidence rate of systemic adverse effects among children aged 5-12 month-old was 1.78% (411/23 113), which was higher than that among children aged 13-24 month-old (1.36%, 154/11 298), the difference showed statistical significance (χ(2) = 8.10, P < 0.01). The incidence rate of local adverse effects in children vaccinated the first dose was 1.72% (536/31 129), which was higher than that in children vaccinated the second or third dose (0.55%, 18/3282), the difference showed statistical significance (χ(2) = 25.81, P < 0.01). The incidence rate of systemic adverse effects in children vaccinated the first dose was 1.73% (539/31 129), which was higher than that in children vaccinated the second or third dose (0.79%, 26/5282), whose difference also showed statistical significance (χ(2) = 16.22, P < 0.01).

CONCLUSIONThe safety of meningococcal group AC bivalent polysaccharide conjugate vaccine among children aged 5-24 months old is relative good.

Female ; Humans ; Infant ; Male ; Meningitis, Meningococcal ; microbiology ; prevention & control ; Meningococcal Vaccines ; administration & dosage ; adverse effects ; immunology ; Neisseria meningitidis, Serogroup A ; Neisseria meningitidis, Serogroup C ; Polysaccharides, Bacterial ; immunology ; Vaccines, Conjugate ; administration & dosage ; adverse effects ; immunology

Objective To investigate the epidemiological characteristics of malaria and implementation of the “1-3-7” approach in malaria elimination in Yunnan Province, so as to provide the data support for the development of post-elimination surveillance interventions. Methods All data pertaining to malaria cases in Yunnan Province from 2014 to 2019 were captured from the Notifiable Disease Reporting System of Chinese Center for Disease Control and Prevention, and the changes in the epidemic situation of malaria were analyzed during the 5-year period. In addition, the core indexes regarding the “1-3-7” approach in malaria elimination of Yunnan Province from 2014 to 2019 were retrieved from the Malaria Control System in the Parasitic Disease Information Reporting System, and all changes in the indexes were descriptively analyzed. Results During the period from 2014 to 2019, a total of 2 283 malaria cases were reported in Yunnan Province, including 1 927 cases with vivax malaria, 326 cases with plasmodium malaria, 29 cases with other species of malaria, and one case with unidentified species. There were 64 local cases, 2 219 overseas imported cases. Among the 2 283 malaria cases, the male/female ratio was 4.58∶1, and 80.25% of the cases were aged from 15 to 50 years. Farmer (70.00%) was the predominant occupation, and 76.70% (1 751/2 283) of the cases were identified in 25 border counties (districts). Malaria cases were reported in each month during the 5-year period, and the number of malaria cases increased from April, peaked on May to July, and started to decline on August. From 2014 to 2019, the reporting rate of malaria cases within 24 hours upon diagnosis was 100%, and the detection of malaria cases was 99.69% (2 276/ 2 283) in the laboratory, with a 99.65% (2 275/2 283) rate of definite diagnosis. In addition, the percentage of individual epidemiological investigations within 3 days was 100.00% (2 283/2 283), and the number of epidemic foci survey and treatment within 7 days was 576 during the 3-year period from 2017 to 2019. The goal of malaria elimination was achieved in Yunnan Province on June, 2020. Conclusions Malaria has been eliminated in Yunnan Province, and management of overseas imported malaria is the primary challenge to consolidate the malaria elimination achievements in the future. However, the approach in malaria elimination remains to be maintained, and the role of the Yunnan Provincial Malaria Diagnostic Reference Laboratory requires to be strengthened.