To explore a new way to treat CML, inhibitory effect of small interfering RNA (SiRNA) on bcr-abl fusion gene expression of K562 cell line was studied. SiRNA for bcr-abl gene was designed and transfected into K562 cells, bcr-abl gene expression was tested by RT-PCR. The results showed that bcr-abl gene expression was inhibited by using siRNA in dose-dependent manner and reduced to 19.9% and 26.6% of the control at 24 and 48 hours after transfection with 0.2 micro g siRNA respectively. K562 cells proliferation was suppressed finally, but bcr-abl gene expression restored at 72 hours. In conclusion, anti-bcr-abl siRNA can effectively inhibit bcr-abl gene expression of K562 cell line.
Dose-Response Relationship, Drug ; Genes, abl ; Humans ; K562 Cells ; metabolism ; RNA, Messenger ; analysis ; RNA, Small Interfering ; pharmacology ; Transfection

Citrus tristeza virus (CTV) was purified from a citrus sample by a modified protocol, and the yield was about 1 mg from 100 g citrus tissues. Polyclonal antibody was prepared by immunizing rabbits with the purified CTV preparation with a titer 1:25600 in indirect ELISA test. Eighteen hybridoma-cell lines secreting monoclonal antibodies (MAbs) against CTV were screened after the fusion of mouse myeloma cells (SP2/0) with spleen cells from BALB/c immunized with the virus preparation. Four hybridoma-cell lines were selected randomly for later analysis. The results indicated that the titers of ascetic fluids against these hybridoma cell lines ranged from 1:51200 to 1:204800 in indirect ELISA, and their isotypes and subclasses were IgG2a for 2G and 3H and IgG2b for IE and 4H. These four Mabs were used to detect CTV in citrus samples in different sources. Results showed that TAS-ELISA with polyclonal antibody as trapping antibody and monoclonal antibody as testing antibody had a higher specificity and sensitivity than PAS-ELISA. Four Mabs showed different intensities of serological reaction with different CTV isolates. However, much work remains for realizing the characteristics and the serological relationships among these isolates.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Citrus ; virology ; Closterovirus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; Mice ; Mice, Inbred BALB C ; Rabbits

Objective To detect the positive rates of antibodies against avian influenza virus (AIV) subtypes H5, H6, H7 and H9 among people in poultry occupations in Guangdong province and to analyze the transmission of various subtypes of AIV from poultry to human contacts for the prevention and control of novel AIV infection in human beings.Methods Serum specimens were collected from 1066 peo-ple in poultry occupations ( occupational group) and 205 people not in poultry occupations ( non-occupational group) in 10 cities of Guangdong province.The inactivated AIV strains, isolated from poultry or environment of Guangdong province, were used as antigens to detect antibodies against AIV subtypes H5, H6, H7 and H9 by using the hemagglutination inhibition ( HI) assay.Results The positive rates of antibodies against AIV subtypes H5, H6, H7 and H9 carried by people from the occupational group were respectively 0.44%, 0%, 0.30%and 0.30%in 2013 and 1.08%, 0.0%, 0.0%and 0.27%in 2014.Only the anti-H9 anti-bodies were detected in serum samples collected form people in the non-occupational group in 2013 with a positive rate of 0.95%.No significant differences with the positive rates of anti-AIV antibodies were found between the occupational group and the non-occupational group.However, the geometric mean titer ( GMT) of anti-AVI antibodies in people from the occupational group was higher than that of the non-occupational group.Conclusion Although a grand spread of AIV from avian to human is not likely to happen yet, con-tacting with poultry is the risk factor for AIV infection in Guangdong population.A long-term surveillance of anti-AIV antibodies in serum should be strengthened among people in poultry occupations for the timely pre-vention and control of novel AIV outbreak.

OBJECTIVETo investigate the cytotoxic effects of mTOR inhibitor rapamycin (Rapa) and idarubicin (IDA) on human T-cell acute lymphoblastic leukemia Jurkat cell line.

METHODSThe proliferation of Jurkat cells was observed by CCK-8 assay. The combined index was analyzed by Isobologram method. Apoptosis was detected by electron microscopy and flow cytometry with Annexin V/PI staining. Protein expressions of Caspase 3, PARP, Caspase 8, Caspase 9, Akt, p-Akt, P85S6K, p-P85S6K, P70S6K, p-P70S6K, ERK1/2 and p-ERK1/2 were determined by Western blotting.

RESULTSThe IC(50) of IDA for Jurkat cells was significantly reduced when combined with 10 nmol/L rapamycin. The combined index was <1. Both electron microscopy and Annexin V/PI staining flow cytometry revealed that rapamycin significantly increased apoptotic sensitivity to IDA. The combination of IDA with rapamycin enhanced the expressions of Caspase 3, PARP, Caspase 8 and Caspase 9. Rapamycin significantly inhibited mTOR signaling upstream Akt and downstream S6K activation triggered by IDA, and the combination of the two agents led to synergistic inhibition of ERK phosphorylation.

CONCLUSIONCombination of IDA with rapamycin exerted a synergistic anti-proliferative effect and promoted apoptosis by both extrinsic and intrinsic apoptotic pathways in Jurkat cells. Inhibition of ERK phosphorylation and mTOR signaling by rapamycin may play a certain role in this synergistic effect.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Drug Synergism ; Humans ; Idarubicin ; pharmacology ; Jurkat Cells ; metabolism ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Sirolimus ; pharmacology

Objective To investigate the effect of ursolic acid (UA) on the colorectal tumor and microenvironment in mice,and to provide a theoretical basis for the clinical application of UA.Methods The models of subcutaneous transplanted tumor of mouse CT26 cells was established.The models were divided into four groups:control group,tumor bearing group,tumor beating dimethyl sulfoxide (DMSO) group and tumor beating UA group.Tbe serum levels of interleukin-6 (IL-6) were detected by enzyme linked immunosorbent assay (ELISA).The number and percentage of myeloid-derived suppressor cell (MDSC) in the spleen of mice were analyzed by flow cytometry.The mRNA levels of IL-6 and signal transducer and activator of transcription 3 (STAT3) in tumor were examined by real-time quantitative polymerase chain reaction (RT-PCR).The protein levels of STAT3 and p-STAT3 in tumor were detected by Western blotting.Results The results showed that UA could significantly decrease the number of spleen MDSC.The accounts of spleen MDSC of tumor bearing UA group (249.60 ± 17.12) was lower than that of tumor beating DMSO group (366.40 ± 34.08),and the difference was statistically significant (P =0.021).The serum level of IL-6 in tumor bearing UA group [(46.40 ± 17.05) pg/ml] was decreased than that in tumor bearing DMSO group [(94.27 ±21.51) pg/ml],and the difference was statistically significant (P =0.012).The expression levels of IL-6 and STAT3 mRNA in tumor tissues of tumor bearing UA group (0.12 ±0.01,0.21 ±0.08) were lower than those of tumor bearing DMSO group (0.69 ± 0.14,0.79 ± 0.06),and the differences were statistically significant (P =0.008;P =0.003).The protein expression levels of STAT3 and p-STAT3 in tumor tissues of tumor bearing UA group (0.81 ±0.02,0.28 ±0.04) were lower than those of tumor bearing DMS0 group (0.98 ±0.02,0.91 ±0.22),and the differences were statistically significant (P =0.027;P =0.029).Conclusion UA may inhibit the activation of STAT3 signaling pathway and the amplification of MDSC in microenvironment by reducing IL-6,thus to enhance the function of immune-killing tumor cells to regulate tumor immune microenvironment and inhibit the immune escape of mouse colorectal cancer cells.

Objective To study the changes of plasma normetanephrine(NMN) and metanephrine(MN) during the resection of adrenal pheochromocytoma. Methods Fourteen patients with adrenal pheocromocytoma and 9 patients with adrenal cortex tumor were recruited in our study. Blood samples were obtained at these time points: after anesthesia induction,the beginning of incision of skin, when exploring the tumor,resection of the tumor, and the end of anesthesia. The NMN and MN were determined by high performance liquid chromatogram (HPLC). Results NMN were obviously different among 5 time points in the patients with adrenal pheocromocytoma (P0.05). No significant difference was found between NMN and MN in the patients with adrenal cortex tumor. Conclusion NMN has markedly changed during the resection of adrenal pheochromocytoma, while MN has been relatively stable. The anesthesia induction and exploring of the tumor are the key of a successful operation. MN is the stable index in the diagnosis of adrenal pheochromocytoma.

OBJECTIVETo evaluate the clinical effects of Fu Fang Ya Tong Ding on treatment of gingivitis and pericoronitis.

METHODS120 clinical patients with gingivitis or pericoronitis were randomly divided into 3 groups (40 patients in each group). After routine rinse treatment for all patients, patients in the test group were treated with Fu Fang Ya Tong Ding, patients in the positive group were treated with iodine glycerol, while that time patients in the negative group received no treatment anymore. Ten minutes after treatment, visual analogue scale (VAS) was used to record the severity of pain for each patient. 3 days and 7 days later, pain and inflammation degree were also recorded by pain three-degree scoring method and index of gingivitis. The total treatment effects were evaluated under a comprehensive clinical treatment standard.

RESULTS10 minutes after treatment, 40.0% of patients in the test group had almost no pain, while no obvious reduction of pain was found in the control group. 3 days, 7 days after the treatment, 92.5%, 95.0% of patients in the test group had no pain, and 55.0%, 90.0% of patients in the positive group had no pain. In the negative group, there were 47.5% of patients which pain was still remained in 7 days. 7 days after treatment, gingival index in the test group reduced by 25.0% and 42.8% compared with the positive and negative groups (P<0.05). 3 days after treatment, 62.5%, 45.0% and 30.0% patients separately in the test, positive and negative groups manifested good effects under the comprehensive clinical treatment standard; after 7 days, 97.5%, 92.5% and 77.5% patients in the 3 groups manifested good effect. The group using Fu Fang Ya Tong Ding had better effects than groups using iodine glycerol or only applying routine rinsing treatment group (P<0.05).

CONCLUSIONFu Fang Ya Tong Ding can treat gingivitis and pericoronitis through significantly reducing inflammation and pain.

Adult ; Female ; Gingivitis ; Humans ; Male ; Mouthwashes ; Pericoronitis ; Periodontal Index

Objective To evaluate the performance of modified Hodge test on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Methods Fortynine Enterobacteriaceae isolates with decreased susceptibility to carbapenems ( MIC of imipenem, meropenem or ertapenem was ≥ 2 μg/ml ) were collected from 16 teaching hospitals from 2004 to 2008. MICs of imipenem, meropenem and etapenem were determined by agar dilution method. Carbapenemases were detected by modified Hodge test. Carbepenemase-causing positive results and AmpCs-causing positive results were differentiated by phenyl boronic acid and oxacillin. Beta-lactamases encoding genes including blaNDM-1were detected by PCR and sequencing. Results Thirty-six of 49 isolates were non-susceptible to imipenem (MIC >4 μg/ml), 31 were non-susceptible to meropenem (MIC > 4 μg/ml) and 47 were non-susceptible to ertapenem (MIC > 2 μg/ml). Twenty-three isolates showed positive modified Hodge test result, including 9 weak-positive results and 14 strong-positive results. Through PCR detection and sequencing, 2 out of 9 isolates showing weak-positive results carried blaKPC-2 and other 7 did not carry any carbapenemase genes but AmpCs/ESBLs genes. Among the 14 isolates showing strong-positive results, 4 carried blaKPC-2, 8 carried blaIMP-4 and 2 caried blaIMP-8. All 26 isolates with negative modified Hodge test result didn't carry any carbapenemase genes. No isolate carried blaNDM-1. Carbapenemases genes PCR detection was regarded as a gold standard, and the sensitivity, specificity, positive predictive value and negative predictive value of modified hodge test was 100%, 79%, 70% and 100% on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Conclusions Modified Hodge test revealed great sensitivity but showed a few false positive results. True and false positive results can be effectively differentiated by phynel boronic acid and oxacillin.

Objective To investigate the susceptibility and resistance of clinical isolates collected from hospitals in several regions of China . Methods Fourteen general hospitals and two children ’ s hospitals were involved in this program . Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated Systems .Results were analyzed according to CLSI 2013 breakpoints .Results A total of 84 572 clinical isolates were collected from January to December 2013 ,of which gram negative organisms and gram positive cocci accounted for 73 .0% and 27 .0%respectively .Methicillin-resistant strains in S .aureus (MRSA) and coagulase negative Staphylococcus (MRCNS) accounted for an average of 45 .2% and 73 .5% respectively .The resistance rates of methicillin-resistant strains to β-lactams and other antimicrobial agents were much higher than those of methicillin-susceptible strains .However ,92 .2% of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole while 87 .4% of MRCNS strains were susceptible to rifampin . No staphylococcal strains were found resistant to vancomycin ,teicoplanin or linezolid .In Enterococcus spp .,the resistance rates of E . f aecalis strains to most tested drugs (except chloramphenicol) were much lower than those of E . f aecium .Some strains of both species were resistant to vancomycin .Vancomycin-resistant strains of E . f aecalis and E . f aecium were mainly VanA type based on their phenotype .Regarding non-meningitis S . pneumoniae strains ,the prevalence of penicillin-susceptible S . pneumoniae and penicillin-intermediate S . pneumoniae strains isolated from both adults and children were lower than those isolated in 2012 ,but the prevalence of penicillin-resistant S .pneumoniae strains increased .The prevalence of ESBLs producing strains was 54 .0% in E .coli ,31 .8% in Klebsiella spp .(K .pneumoniae and K .oxytoca) and 16 .5% in Proteus mirabilis isolates on average . ESBLs-producing Enterobacteriaceae strains were more resistant than non-ESBLs-producing strains in terms of antibiotic resistance rates .The strains of Enterobacteriaceae were still highly susceptible to carbapenems .Overall less than 7 .0% of these strains were resistant to carbapenems .About 62 .8% and 59 .4% of Acinetobacter spp .(A .baumannii accounts for 89 .2% ) strains were resistant to imipenem and meropenem ,respectively .Compared with the data of year 2012 , extensively-drug resistant strains in K . pneumoniae and A . baumannii decreased .Conclusions The antibiotic resistance of clinical bacterial isolates is growing in 2013 .The disseminated multi-drug or pan-drug resistant strains in a special region poses a serious threat to clinical practice and implies the importance of strengthening infection control .

Objective To analyze the resistance proifle of bacterial strains isolated from geriatric patients in 17 hospitals across China from 2005 to 2014.Methods A total of 17 representative general hospitals were involved in this program. Bacterial susceptibility testing was carried out by means of a uniifed protocol using Kirby-Bauer method and MIC determination. The results were analyzed according to CLSI 2014 breakpoints.Results The proportion of the strains isolated from geriatric patients among all the clinical isolates increased with time from 30.0% in 2005 to 32.7% in 2014. A total of 159 888 clinical isolates were collected from geriatric patient during the period from 2005 to 2014, about 33.1% of the whole patient population. Gram negative organisms and gram positive cocci accounted for 77.1% (123 229/159 888) and 22.9% (36 659/159 888), respectively. Majority (92.8%, 148 376/159 888) of the isolates were from inpatients and more than half (55.2%, 88 201/159 888) of the isolates were from sputum or other respiratory tract specimens. Methicillin-resistant strains inS. aureus (MRSA) and coagulase negativeStaphylococcus (MRCNS) accounted for an average of 67.1% and 75.9%, respectively. The resistance rates of methicillin-resistant strains to β-lactams and other antimicrobial agents were much higher than those of methicillin-susceptible strains. No staphylococcal strains were found resistant to vancomycin, teicoplanin or linezolid. Some strains ofE. faecalis (0.4%) andE. faecium (4.6%) were resistant to vancomycin, which was higher than average national level (0.3%, 3.2%). Vancomycin-resistant strains ofE. faecalisandE. faecium were mainly VanA type and VanB type based on their phenotype. The prevalence of penicillin-susceptibleS. pneumoniae strains was 78.2%, slightly lower than the 95.0% in Chinese adults in year 2014. The prevalence of ESBLs-producing strains was 67.5% inE. coli, 40.4% inKlebsiella (K. pneumoniae andK. oxytoca) and 34.3% inProteus mirabilis isolates on average. The strains ofEnterobacteriaceae were still highly susceptible to carbapenems (<10% resistant), followed by amikacin and the beta-lactam and beta-lactamase inhibitor combinations. Overall, 35.9% and 33.0% of theP. aeruginosa strains were resistant to imipenem and meropenem. More than 50% of theA. baumannii strains were resistant to imipenem and meropenem. The prevalence of extensively drug-resistant (XDR)P. aeruginosa (4.0%-1.8%) was higher than the average national level (2.1%-1.6%). The prevalence of XDR A. baumannii (19.2%-15.5%) and XDREnterobacteriaceae (0.1%-1.0%) was lower than the average national level (21.4%-19.7% and 0.3%-3.2%).Conclusions The proportion of clinical isolates from geriatric patients is different from average national level at the same period. The isolates from geriatric patients are more likely from inpatients, respiratory tract specimens and more likely non-fermentative gram-negative bacilli compared to average national level. The proportion of fastidious bacteria andEnterobacteriaceae species is generally lower than average national level. MRSA, VRE, ESBLs-producing strains and XDRP. aeruginosa are more prevalent in geriatric patients than in general Chinese patient population. This study suggests that surveillance of antimicrobial resistance for the clinical isolates from geriatric patients is very important for rational antimicrobial therapy.