Objective To investigate the clinical significance of peripheral blood parameters including immature reticulocyte fraction (IRF) in monitoring the hematopoietic recovery of patients with acute myeloid leukemia aftcr receiving consolidation chemotherapy with high dose cytarabine (HDAC).Methods The peripheral blood parameters,including absolute neutrophil (ANC) and platelet (Plt) counts as well as IRF,were measured by Sysmex XE-2100 automated hematology analyzer before and after consolidation chemotherapy with HDAC in 30 patients with acute myeloid leukemia.These peripheral blood parameters were further statistically analyzed and compared.Results Following consolidation chemotherapy,the median time to recovery for ANC,Plt and IRF was (9.2±2.5) days,(11.3±3.7) days and (5.0±2.6) days,respectively.In comparison with ANC and Plt recovery,IRF showed significantly early recovery (P ＜ 0.05).Additionally,ANC raised preceding Plt recovery (P ＜ 0.05).However,the median recovery time of ANC,Plt and IRF showed little difference among these patients further subgrouped according to FAB and risk based classification (P ＞ 0.05).Conclusion IRF is an early indicator for the hematopoietic recovery of patients with acute myeloid leukemia following consolidation chemotherapy with HDAC,which could provide important clinical information to evaluate therapeutic efficacy.

Clarithromycin ; therapeutic use ; Humans ; Lymphoma, B-Cell, Marginal Zone ; drug therapy ; Male ; Middle Aged ; Tomography, X-Ray Computed

OBJECTIVE:To obverse the efficacy and safety of tolynicate and napthylacetic acid and atorvastation combined with Danshen injection in the treatment of alcoholic hyperlipemia syndrome. METHODS:21 patients with alcoholic hyperlipemia syndrome were enrolled. All patients were given maintaining acid-base balance,oxygen inhalation and other conventional treatment. Based on it,they were orally given 75 mg Tolynicate and napthylacetic acid tablet,3 times a day+20 mg Atorvastatin calcium tab-let,once a day+4 ml Compound danshen injection,adding into 150 ml 5% Glucose injection by intravenous infusion,once a day. The treatment course for both groups was 30 d. Clinical efficacy,ALT,AST,TBIL,TC,TG,RBC and Hb before and after 6, 12,18,24 and 30 d,and incidence of adverse reactions were observed. RESULTS:After treatment,the total effective rate was 95.3%and incidence of adverse reactions was 28.5%. The ALT,AST and TBIL after 6,12,18,24 and 30 h and TC and TG after 18,24 and 30 h were significantly lower than before and gradually decreased by time;RBC and Hb after 12,18,24 and 30 h were significantly higher than before and gradually increased by time,the differences were statistically significant(P<0.05). CON-CLUSIONS:tolynicate and napthylacetic acid and atorvastation combined with Danshen injection is effective in the treatment of al-coholic hyperlipemia syndrome,with good safety.

Objective To clone and express bradyzoite antigen 1(BAG1) gene of T. gondii,and analyze the immunoreactivity of the recombinant product. Methods The differentiation of T. gondii RH strain tachyzoites into bradyzoites was induced in vitro,and the coding sequence of BAG1 was amplified from bradyzoites by RT-PCR. The PCR product was analyzed by sequencing. The BAG1 coding sequence was further subcloned into the plasmid pET32a(+). The plasmid pET32a(+)-BAG1 was then transformed into BL21(DE3) to express after IPTG induction. The expression product was purified with Ni-NTA agarose and the purified BAG1 was further analyzed by Western blotting and ELISA. Results BAG1 cDNA was amplified from bradyzoites. After IPTG induction,BAG1 was expressed in a fusional form in E. coli. Western blotting showed that the purified recombinant protein could be specifically recognized by sera from mice chronically infected by T. gondii B36 strain. ELISA showed that the positive rate of T. gondii IgG antibodies of 350 human sera detected by the recombinant BAG1(17.4%) was higher than by recombinant SAG1 (12.6%)(P

9α-hydroxy-4-androstene-3,17-dione (9-OH-AD) is an important intermediate in the steroidal drugs production. 3-ketosteroid-9α-hydroxylase (KSH), a two protein system of KshA and KshB, is a key-enzyme in the microbial steroid ring B-opening pathway. KSH catalyzes the transformation of 4-androstene-3,17-dione (AD) into 9-OH-AD specifically. In the present study, the putative KshA and KshB genes were cloned from Mycobacterium smegmatis mc(2)155 and Gordonia neofelifaecis NRRL B-59395 respectively, and were inserted into the expression vector pNIT, the co-expression plasmids of kshA-kshB were obtained and electroporated into Mycobacterium sp. NRRL B-3805 cells. The recombinants were used to transform steroids, the main product was characterized as 9α-hydroxy-4-androstene-3,17-dione (9-OH-AD), showing that kshA and kshB were expressed successfully. Different from the original strain Mycobacterium sp. NRRL B-3805 that accumulates 4-androstene-3,17-dione, the recombinants accumulates 9α-hydroxy-4-androstene-3,17-dione as the main product. This results indicates that the putative genes kshA, kshB encode active KshA and KshB, respectively. The process of biotransformation was investigated and the results show that phytosterol is the most suitable substrate for biotransformation, kshA and kshB from M. smegmatis mc(2)155 seemed to exhibit high activity, because the resultant recombinant of them catalyzed the biotransformation of phytosterol to 9-OH-AD in a percent conversion of 90%, which was much higher than that of G. neofelifaecis NRRL B-59395. This study on the manipulation of the ksh genes in Mycobacterium sp. NRRL B-3805 provides a new pathway for producing steroid medicines.
Androstenedione ; analogs & derivatives ; biosynthesis ; Bacterial Proteins ; genetics ; metabolism ; Biotransformation ; Ketosteroids ; Mixed Function Oxygenases ; genetics ; metabolism ; Mycobacterium ; metabolism ; Mycobacterium smegmatis ; enzymology ; Plasmids

Serum folate and vitamin B12 were measured in 171 healthy children under 14 years of age, 30 normal adults and 30 samples of normal cord blood. All the 171 children had been supplied with adequate folic acid and vitamin B12 before measurements. The results -were: 1. The lower limits of normal serum folate values were 6.1 nmol/L under 4 years, 8.4 nmol/L at age of 4-14 years and 5.0 nmol/L in adults. 2. The lower limits of normal serum vitamin B12 values were 459 pmol/L under 1 year and 107 pmol/L at age of 1 year to adults.

AIM:To investigate the effect of small interfering RNA(siRNA)-mediated progranulin(PGRN) gene silencing on the proliferation and migration abilities of human non-small-cell lung cancer A549 cells and its mecha-nism.METHODS:The mRNA and protein expression levels of PGRN in the A 549 cells and human bronchial epithelial (HBE)cells were detected by qPCR and Western blot.A549 cells were transfected with PGRN-siRNA by liposome meth-od.The expression of PGRN at mRNA and protein levels in the A 549 cells transfected with PGRN-siRNA was detected by qPCR and Western blot, respectively.The cell viability was measured by MTT assay.The cell proliferation ability was measured by living cells counting and crystal violet staining assays.The cell migration ability was measured by wound-heal-ing and Transwell assays.The protein levels of proliferating cell nuclear antigen(PCNA),cyclin D1,Bcl-2 and Bax were determined by Western blot.The protein levels of phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2) and phosphorylated protein kinase B(p-Akt)were also determined by Western blot.RESULTS:The expression of PGRN at mRNA and protein levels was higher in the A 549 cells than that in the HBE cells(P<0.05).The expression of PGRN at mRNA and protein levels in the A549 cells transfected with PGRN-siRNA was significantly decreased,and the cell pro-liferation and migration abilities were significantly decreased.The protein expression levels of PCNA,cyclin D1 and Bcl-2 were significantly reduced and the protein expression level of Bax was significantly increased(P<0.05).Meanwhile,the protein levels of p-ERK1/2 and p-Akt were down-regulated(P<0.05).CONCLUSION:PGRN gene silencing obviously inhibits the proliferation and migration abilities of human non-small-cell lung cancer A549 cells.The PI3K/Akt and MAPK/ERK signaling pathways may play an important role in these processes.

OBJECTIVETo investigate the effects of survivin antisense oligodeoxynucleoties (ASODN) transfection mediated by cytofectin on apoptosis and cisplatin resistance in cisplatin resistant human lung adenocarcinoma A549/CDDP cells in vitro.

METHODSA549/CDDP cells were cultured routinely in RPMI-1640 medium. Survivin ASODN mediated by cytofectin was transfected into the A549/CDDP cells. Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry SABC assays were performed to determine the regulation of survivin expression by ASODN. The influence of ASODN transfection on apoptosis was determined by fluoroscence microscopy and Hoechst staining, agarose gel electrophoresis, flow cytometry and caspase-3 colorimetric assay. MTT assay was performed to detect the cell viability, half-maximum inhibitory concentration (IC50) and cisplatin resistance index (RI) were thereby calculated.

RESULTSTransfected by survivin ASODN for 48 h, down-regulation of survivin expression was measured, of which mRNA and protein expression was significantly down-regulated to 41.56% and 0.864 +/- 0.045, respectively (P < 0.05). Transfection with survivin ASODN caused typical apoptotic changes, including characteristic chromatin condensation, nuclear shrinkage, nuclear cleavage and the cells grew more regularly, and some cells were floating. Typical DNA ladder pattern was observed by agarose gel electrophoresis. Furthermore, apoptotic index and caspase-3 activity was enhanced to 34.03% and 1.1298 +/- 0.2502, respectively (P < 0.05). It was significantly different as compared with the control group. While combination with ASODN and 10 micromol/L cisplatin caused far more distinctive apoptotic alterations, of which AI and caspase-3 activity reached to 65.85% and 1.6805 +/- 0.2758, respectively (P < 0.05), and even compared with the single ASODN group, the difference was still significant (P < 0.05). Transfected with survivin ASODN only or with combination of cisplatin for 48 h, the inhibitory rate of cell growth was enhanced to 59.3% and 83.7% (P < 0.05), respectively, while inversely, the cell viability reduced to a lowest value. The half-maximum inhibitory concentration of cisplatin was reduced from 225.03 +/- 10.59 micromol/L to 158.84 +/- 4.26 micromol/L, and the resistant index was conversely reduced from 11.9 to 8.39. Non-sense oligodeoxynucleotides (NSODN) and liposome had no effect on the cells growth (P > 0.05).

CONCLUSIONTransfection with survivin ASODN can to a great extent reverse the cisplatin resistance in human cisplatin resistant lung adenocarcinoma cells A549/CDDP in vitro, and can thereby significantly inhibit the growth of the cells. The mechanism of reversal of resistance to cisplatin by this transfection can be associated with specific down-regulation of survivin expression, which decreases the threshold of apoptosis, induces more pronounced apoptosis,and reverses the resistance to apoptosis induced by cisplatin in A549/CDDP cells in vitro.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Down-Regulation ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Microtubule-Associated Proteins ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

The samples of sulfur-fumigated Paeoniae Alba Radix acquired both by random spot check from domestic market and self-production by the research group in the laboratory were used to evaluate the effects of sulphur fumigation on the quality of Paeoniae Alba Radix by comparing sulfur-fumigated degree and character, the content of paeoniflorin and paeoniflorin sulfurous acid ester, and changes of the fingerprint. We used methods in Chinese Pharmacopeia to evaluate the character of sulfur-fumigated Paeoniae Alba Radix and determinate the content of aulfur-fumigated paeoniflorin. LC-MS method was used to analyze paeoniflorin-converted products. HPLC fingerprint methods were established to evaluate the differences on quality by similarity. Results showed that fumigated Paeoniae Alba Radix became white and its unique fragrance disappeared, along with the production of pungent sour gas. It also had a significant effect on paeoniflorin content. As sulfur smoked degree aggravated, paeoniflorin content decreased subsequently, some of which turned into paeoniflorin sulfurous acid ester, and this change was not reversible. Fingerprint also showed obvious changes. Obviously, sulfur fumigation had severe influence on the quality of Paeoniae Alba Radix, but we can control the quality of the Paeoniae Alba Radix by testing the paeoniflorin sulfurous acid ester content.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; methods ; Paeonia ; chemistry ; Quality Control ; Sulfur ; chemistry

ObjectiveTo estimate the clinical efficacy and immunomodulatory effect of polysaccharidenucleic acid fraction of bacillus calmette guerin(BCG-PSN) combined with a traditional Chinese medicine in patients with vitiligo.MethodsThis study recruited 99 patients with vitiligo aged from 13 to 65(35,6 ± 5.8) years.The patients were classified into 3 groups to be treated with BCG-PSN and a traditional Chinese medicine (Baidianfeng granules) alone or in combination.BCG-PSN was intramuscularly injected at a dose of 2 ml every other day and baidianfeng granules were given orally thrice a day,for 3 months.Peripheral blood samples were obtained from the patients at the baseline and after the end of treatment and from 30 healthy controls.Flow cytometry and enzyme linked immunosorbent assay(ELISA) were performed to detect T cell subsets and expression level of Fas and Fas ligand(FasL),respectively.Data were analyzed by t test and chi-square test.Results The response rate was significantly higher in patients treated with BCG-PSN combined with Baidianfeng granules than in those with BCG-PSN alone(82.86％ vs.40.63％,P ＜ 0.01 ).Before the treatment,patients showed a lower percentage of CD3+ cells,CD3+CD4+ cells and CD3+CD8+ cells in peripheral blood(all P ＜0.01 ),weaker expression of Fas (P ＜ 0.01 ),but a higher CD4/CD8 ratio (P ＜ 0.01 ) compared with the controls.The treatment with BCG-PSN and Baidianfeng granules alone or in combinatiou all induced an increase in the percentage of CD3+ cells,CD3+CD4+ cells and CD3+CD8+ cells in peripheral blood(P ＜ 0.05) and in the expression of Fas(P ＜ 0.01),but a decrease in CD4/CD8 ratio(P ＜ 0.05).ConclusionsBCG-PSN may induce the normal apoptosis in lymphocytes via reversing the abnormality in the expression of Fas/FasL by peripheral blood lymphocytes,and the effect of BCG-PSN may be enhanced by a traditional Chinese medicine,Baidianfeng granules.