Objective To determine the acetylation level of nucleophosmin(NPM)in female breast cancer and to discuss its function through mutation of modified lysine sites.To construct positive and negative NPM mutants on its acetylated lysine sites and to express them in breast cancer cells.Methods Acetylation level and acetylated lysine sites of NPM in three breast cancer tissues and para-carcinoma tissues were detected by acetylome technology;NPM mutants were constructed by site-directed mutagenesis PCR,specific PCR products were digested by DpnI and transformed into Escherichia coli(E.coli)to obtain specific plasmids for mutants;The accuracy of mutants were verified by double restriction enzyme digestion and sequencing;The mutants were expressed in BT-549 cells by transient transfection and verified by RT-PCR method.Protein expression and acetylation level of NPM were validated by Western blotting;Function of NPM acetylation was analyzed by proteomic detection and bioinformatic analysis.Results The 27th and 32nd lysine of NPM were highly acetylated in breast cancer tissues,which were 2.76 and 2.22 times higher than those in adjacent normal tissues,respectively;The NPM mutants showed the same molecular weight as that of wild type NPM and contained expected mutation sites;Corresponding NPM mRNA levels of BT-549 cells transfected with NPM mutants were significantly increased.With the increase of wild type NPM expression level,NPM acetylation level increased,while decreased after 27th lysine underwent negative mutation.NPM acetylation can significantly change the expression levels of 101 proteins in BT-549 cells,which are enriched in regulation of cellular macromolecule biosynthesis,DNA-template transcription,RNA biosynthesis and RNA metabolism process.Conclusion NPM is highly acetylated in breast cancer and can play a key role in cellular macromolecule biosynthesis,DNA-templated transcription,RNA biosynthesis and RNA metabolism process.

OBJECTIVE: To compare the therapeutic effect of Tongdu Tiaoqi acupuncture (acupuncture for unblocking governor vessel and regulating qi ) combined with warming acupuncture, Tongdu Tiaoqi acupuncture, abdominal moxibustion and oral tamsulosin hydrochloride sustained release capsule on postoperative urinary retention. METHODS: A total of 120 patients with postoperative urinary retention were randomized into an acupuncture-moxibustion group, an acupuncture group, a moxibustion group and a medication group, 30 cases in each group. Tongdu Tiaoqi acupuncture combined with warming acupuncture were applied in the acupuncture-moxibustion group. Tongdu Tiaoqi acupuncture was applied at Baihui (GV 20), Shuigou (CV 26) etc. in the acupuncture group. Moxibustion was applied at Qihai (CV 6), Guanyuan (CV 4), Shuidao (ST 28) and Sanyinjiao (SP 6) in the moxibustion group. Tamsulosin hydrochloride sustained release capsule was given orally in the medication group. The treatment was once a day, and 5-day treatment was required in each group. Before and after treatment, the residual urine volume of bladder, the visual analogue scale (VAS) score and the time of first urethral catheter removal were observed, and the clinical efficacy was compared in the 4 groups. RESULTS: After treatment, the residual urine volume of bladder was decreased compared before treatment in the 4 groups (P<0.05), and that in the acupuncture-moxibustion group was less than the other 3 groups (P<0.05). After treatment, the VAS scores were decreased compared before treatment in the acupuncture-moxibustion group, the acupuncture group and the moxibustion group (P<0.05), and those in the 3 groups were lower than the medication group (P<0.05). The time of first urethral catheter removal in the acupuncture-moxibustion group was earlier than the other 3 groups (P<0.05). The total effective rate was 93.3% (28/30) in the acupuncture-moxibustion group, which was superior to 63.3% (19/30) in the acupuncture group, 60.0% (18/30) in the moxibustion group and 66.7% (20/30) in the medication group (P<0.05). CONCLUSION The therapeutic effect of Tongdu Tiaoqi acupuncture combined with warming acupuncture on postoperative urinary retention is superior to simple acupuncture, abdominal moxibustion and tamsulosin hydrochloride sustained release capsule.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Moxibustion ; Treatment Outcome ; Urinary Retention/therapy*

To illustrate the intrinsic dissolution rate(IDR) involved in biopharmaceutics classification system of Chinese materia medica(CMMBCS), investigate the effect of artificial multicomponent environment on IDR of berberine, by using a non-disintegrating disk, and summarize related rules by using the obtained data. Progressive levels design was used to investigate the effects of single component environment (puerarin, baicalin, and glycyrrhizic acid); double-component environment(puerarin+baicalin, puerarin+ glycyrrhizic acid, baicalin+glycyrrhizic acid); and triple-component environment(puerarin+baicalin+glycyrrhizic acid) on IDR of berberine, laying a foundation for further researches on the absorption mechanism of multiple components.

OBJECTIVETo explore the influence of FLT3-ITD mutation and ITD length on the overall survival (OS) and relapse free survival(RFS) in patients with non-M3 acute myeloid leukemia.

METHODSClinical features and therapeutic effect were retrospectively analyzed in 75 AML patients with FLT3-ITD mutation and 76 FLT3-ITD AML patients with a normal karotype from June 2011 to April 2016. Genomic DNA was amplified by PCR, and FLT3-ITD mutation length was analyzed by DNA sequencing in 40 patients.

RESULTSAML patients with FLT3-ITD mutation had higher WBC count and the ratio of BM blast cells at initial diagnosis was also higher than those in AML patients without FLT3-ITD mutation (95.13 vs 10.85)(P<0.01); 72% vs 59%(P<0.01). The CR rates in AML patients with FLT3-ITD mutation less than those in AML patients without FLT3-ITD mutation(70.42% vs 94.7%)(P<0.01). OS (P<0.01) and RFS (P<0.01) were significantly increased in patients with AML who received allo-HSCT as compared with the patients who received consolidation chemotherapy and similar to AML patients without FLT3-ITD mutation who received HSCT. Patients with maintenance sorafenib after HSCT had longer OS (P<0.05) and RFS (P<0.05) than controls. ITDs exceeding 60 bp in length were associated with decreasing OS as compared with shorter ITD in AML patients with FLT3-ITD mutation (P<0.05). OS and RFS were similar among the 2 groups receiving consolidation chemotherapy. Besides, the patients with allo-HSCT had shorter ITDs and longer OS than ITDs exceeding 60 bp (P<0.05) and similar to AML patients without FLT3-ITD mutation.

CONCLUSIONAML patients with FLT3-ITD mutation has poorer outcome, among which the prognosis was worse in patients with ITD exceeding 60 bp, and the chemotherapy alone can not improve the prognosis of FLT3-ITD. Allo-HSCT is an effective treatment for AML patients with FLT3-ITD mutation; Sorafenib appears to be an effective maintenance therapy after allo-HSCT in FLT3-ITD AML.

Objective To analyze the possibilities of screening SPF rabbits and guinea pigs from conventional animals, viral antibodies of the conventional rabbits and guinea pigs bred by licensed companies in Guangdong province during 2014-2016 were determined according to the standard of SPF animals in GB14922. 2. Methods Nine batches of 167 rabbit sera and 155 guinea pig sera were sampled from 6 companies. Serum antibodies to virus were determined by ELISA according to GB14922. 2. Results Positivity of antibody to rabbit hemorrhagic disease virus (RHDV) was 82. 2%(129/157) for the vaccinated rabbits, and negative result were obtained for unvaccinated rabbits. Positive rate of rabbit rotavirus (RRV) was 42. 5% (71/167). No positive antibody responses to Sendai virus were detected out in all rabbits. The positive rates of guinea pig reovirus type III (REO-3) and pneumonia virus of mice (PVM) were 52. 9%(82/155)and 20% (31/155) respectively. Antibody responses to Sendai virus ( SV) and lymphocytic choriomeningitis virus ( LCMV) were negative in all guinea pigs. Conclusions Although the conventional rabbits and guinea pigs could meet the national standard, higher infection rates of virus excluded that SPF animals emerged in conventional animals, indicating that selection of SPF animals from conventional colonies is impracticable.

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine

Objective To understand the status of health literacy of residents in Qiandao Lake town in order to complete the surveillance system.Methods A total of 484 residents aged 15 to 69 years old from 9 communities were selected using cluster random sampling method.Then the household interview was conducted.Results The overall level of health literacy of residents was 26.03%.As for the 4 health literacy related aspects including basic health knowledge,health lifestyle and behaviors,health related skill and health beliefs,the rates were 32.54% ,4.77% ,60.30% and 92.62%,respectively. The multivariate logistic regression analysis showed that the level of health literacy of residents was related to age and education.Conclusion The health education in Qiandao Lake town has got some achievements.

This study was aimed to construct the targeting AATF shRNA eukaryotic expression vector and establish the stably transfected U937 cell lines. The sequence of AATF mRNA was obtained from GenBank. After excluding homology, three plasmid expression vectors coding shRNA targeting 228 ∼ 249, 303 ∼ 324 and 443 ∼ 464 of AATF gene sequence were synthesized. Two terminals of shRNA carried BamHI and HindIII restriction sites. The selected nucleotides were cloned into the plasmid pSilencer 3.1-H1 neo respectively, and the resultant recombinant plasmids were named as pSA-1, pSA-2, pSA-3. The sequences of the recombinant plasmids were identified by DNA sequencing. The recombinant plasmids were transfected into the cell line U937 by electroporation with Neon(TM) Transfection System. The transfected cells were persistently screened under G418 (500 mg/L), and isolated with a limited dilution for 8 weeks. The inhibition of AATF mRNA and protein expression was respectively detected by RT-PCR and Western blot. The results indicated that RNAi eukaryotic expression vectors targeting AATF had correct reading frame and nucleotide sequence. Real-time PCR revealed that AATF shRNA effectively silenced mRNA expression of AATF. Western blot analysis found that AATF shRNA obviously suppressed protein expression of AATF (P < 0.05). It is concluded that the shRNA eukaryotic expression vector has been successfully constructed which can inhibit the expression of AATF, and the establishment of stably transfected U937 cell lines provide a original route for exploring the mechanism of AATF in human Leukemia further.
Apoptosis Regulatory Proteins ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Repressor Proteins ; genetics ; Transfection ; U937 Cells

Advanced technologies are used to clarify the meridian tropism theory of traditional Chinese medicine is an important part of theoretical studies of traditional Chinese medicine. In this article, modern pharmacokinetic method was used to investigate tissue distribution characteristics of psoralen and isopsoralen of Psoraleae Fructus decoction in rats, in order to provide research ideas and experimental basis for the meridian tropism theory. In this study, various tissue samples such as heart, liver, spleen, lung, kidney, brain and spermary were collected at different times after oral administration with FP decoction, in order to determine concentration of psoralen and isopsoralen by HPLC. Pharmacokinetic parameters were calculated by DAS 2.0 software. The study results showed that HPLC indexes of psoralen and isopsoralen in various tissues of rats met the determination requirements of biological samples. Both components were distributed in all of the tissues, with AUC(0-t) order of liver > lung approximately kidney > heart > brain approximately spleen > spermary. There was significant difference between liver, kidney, lung and other tissues (P < 0.05). MRT(0-t) of both psoralen and isopsoralen were about 10 h. Therefore, psoralen and isopsoralen showed stronger targeting selection in liver, kidney and lung.
Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Fruit ; chemistry ; Furocoumarins ; pharmacokinetics ; Kinetics ; Male ; Plant Extracts ; administration & dosage ; chemistry ; pharmacokinetics ; Psoralea ; chemistry ; Rats ; Tissue Distribution

The aim of this study was to detect the expression of interleukin-22 (IL-22) and relative CD4(+) T cell subsets in untreated adult patients with immune thrombocytopenia (ITP), and to explore their roles in the pathogenesis of ITP. Fourty adult active ITP patients were enrolled in this study and 40 healthy subjects were taken as control. Plasma IL-22 level was quantified by ELISA. The percentages of Th1, Th17 and Th22 cells in peripheral blood were determined by flow cytometry. The Pearson correlation test was used to evaluate correlations between IL-22 level and Th1 cells, Th17 cells and Th22 cell percentages. The results showed that the plasma IL-22 levels in untreated ITP patients [(364.12 ± 94.22) pg/ml] were significantly higher than that in healthy controls (P < 0.001). The percentages of both Th1 [(18.92 ± 6.03)%] and Th22 [(2.28 ± 0.51)%] cells in ITP patients were elevated as compared to healthy controls (P < 0.05). Elevated plasma concentration of IL-22 positively correlated to the percentages of Th1 (r = 0.42, P = 0.022) and Th22 (r = 0.40, P = 0.030) cells in untreated ITP patients. The percentage of Th17 cells was not significantly different between untreated patients and normal controls, and there was no statistical correlation between the IL-22 level and the percentage of Th17 cells in active ITP patients. It is concluded that elevated IL-22 level correlates to Th1 and Th22 cells percentage, which may play a synergistic effect in the immunopathogenesis of ITP, while Thl7 cells may not be associated with the occurrence of active ITP.
Adolescent ; Adult ; Aged ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; Case-Control Studies ; Female ; Humans ; Interleukins ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; metabolism ; Th1 Cells ; Th17 Cells ; Young Adult