Objective:To study the clinical effects of Yishen-Shengxue Decoction on the prevention and treatment of bone marrow suppression in acute myeloid leukemia after chemotherapy. Methods:A total of 60 patients in Beijing Longfu Hospital from June 2018 to March 2020 were randomly divided into control group and observation group, 30 cases in each group. The control group was treated with western medicine and blood transfusion, while the observation group was treated with Yishen-Shengxue Decoction on the basis of the control group. Both groups were treated for 2 weeks. We recorded the occurrence time and duration of Ⅳ degree myelosuppression of the two groups, compared the score of symptoms and signs, injection volume of recombinant human granulocyte colony stimulating factor (G-CSF) and blood transfusion volume, and recorded the incidence of adverse reactions after chemotherapy. Results:The occurrence of myelosuppression in the observation group was significantly later than that of the control group (5.07 ± 0.87 d vs. 3.83 ± 1.15 d; t=4.695, P<0.01), and the duration of grade Ⅳ myelosuppression was significantly shorter than that of the control group (7.20 ± 0.76 d vs. 10.03 ± 1.30 d; t=10.305, P<0.01); The quantity of granulocyte colony stimulating factor injection in the observation group was significantly less than that of the control group (7.2 ± 0.8 vs. 10.0 ± 1.3, t=10.305), and the quantity of red blood cell suspension (2.5 ± 1.5 U vs. 4.7 ± 1.5 U, t=7.749) and platelet transfusion (1.7 ± 0.5 U vs. 3.1 ± 0.9 U, t=5.879) were significantly less than that of the control group ( P<0.01); the quantitative score of symptoms and signs in the observation group were significantly lower than thoseof the control group ( t values were 18.208, 15.129, respectively, all Ps<0.01). The adverse reactions of the patients in the observation group after chemotherapy were significantly less than those of the control group, and the incidence of infection, bleeding and ECG abnormalities were statistically significant ( χ2 values were 7.500, 10.000, 4.286, respectively, all Ps<0.01). Conclusion:Yishen-Shengxue Decoction is helpful to delay the occurrence of myelosuppression, and promote its recovery, reduce various adverse reactions during myelosuppression, and improve the quality of life.

Department of Pediatrics, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. zhuchuanlong@jsph.org.cn.
Adolescent ; Child ; Female ; Glycyrrhizic Acid ; therapeutic use ; Humans ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Tablets

Objective To investigate the polymorphism of human cytomegalovirus (HCMV) UL144 gene of the low passage clinical isolates in Guangzhou and explore the role of UL144 gene in HCMV pathogenicity. Methods The clinical isolates of HCMV were obtained from the urine sample collected from those infants with intra-uterus HCMV infection in Guangzhou. The virus genome DNA was extracted. According to the genome sequence of Toledo, primers for UL144 gene were designed and used to amplify the complete open reading frames (ORF) of the UL144 gene in our 3 different clinical isolates. These ORFs of the UL144 gene were cloned into pMD18-T vector and their sequences were confirmed by sequencing. Bioinformatics methods were used subsequently to analyze the polymorphisms of these genes in different stains. Results Three HCMV low passage clinical isolates were successfully isolated, named D2, D3 and D52. As shown by PCR, all of these three strains contained UL144 ORF region. Three complete ORFs were amplified in total and their sequences were submitted to GenBank (Accession No.: DQ180368, DQ180382 and DQ180355). In D2, D3 and D52 isolates, their UL144 ORFs consisted of 531 nucleotides. DNA sequences were quite conservative,all variability were base substitution, and the amino acid sequences were high conservative, the rate of amino acid variability was 1.1%. There were no additional or deleted sites of posttranslational modification of UL144 protein in all clinical isolates. There were some differences in the secondary structure among different isolates. The isoelectric point of UL144 protein of all clinical isolates was 8.97. Conclusions All DNA and deduced amino acid sequences of UL144 gene share great similarity among Guangzhou HCMV clinical strains regardless of their polymorphism. It implies that maybe UL144 gene plays an important role in congenital infection.

Objective To evaluate the efficacy and safety of non-steroidal anti-inflammatory drugs (NSAIDs) in preventing heterotopic ossification after hip arthroscopy.Methods Literature search was conducted in PubMed,Embase,Cochrane Library,CNKI and Wanfang data with time range from January 1973 to November 2017.Clinical case control articles on NSAIDs in preventing heterotopic ossification after hip arthroscopy were screened based on the inclusion and exclusion criteria.Meta analysis was done using RevMan 5.3 software to investigate the incidence of complications such as heterotopic ossification and gastrointestinal bleeding after hip arthroscopy in patients taking NSAIDs orally.Results Six articles were included in the study,with a total of 754 cases and 536 controls.NSAIDs reduced the incidence of heterotopic ossification after hip arthroscopy (RR =0.09,95％ CI 0.03-0.27,P ＜ 0.05).Selective COX-2 inhibitor celecoxib (RR =0.17,95％ CI 0.03-0.91,P ＜ 0.05) and PG synthase inhibitor of naproxen (RR =0.17,95％ CI 0.09-0.32,P ＜ 0.05) were also effective in preventing heterotopic ossification.There was no significant difference in the incidence of gastrointestinal complications between the cases and controls after NSAIDs prophylaxis (RR =2.17,95％ CI 0.92-5.12,P ＞ 0.05).Conclusion NSAIDs can effectively reduce the incidence of heterotopic ossification after hip arthroscopy and does not increase the incidence of postoperative gastrointestinal complications.Therefore,it is effective and safe to use NSAIDs to prevent the occurrence of heterotopic ossification after hip arthroscopy.

OBJECTIVETo assess the efficacy of acupuncture for glaucoma.

METHODSThe search was conducted through database to identify randomized controlled trials of acupuncture for glaucoma until September 2010. The quality assessment, data extraction and Meta-analysis were performed by Cochrane Handbook for Systematic Re views of Interventions.

RESULTSEight articles were included. Meta-analysis showed that acupuncture did not decrease intraocular pressure compared with eye drops [SMD = -0.1 66, 95% CI (-1.45, 0.13)]. However, acupuncture increased the effectiveness rate of treatment for glaucoma [OR = 4.45, 95% CI (1.96,10.09)]. Compared with placebo, acupuncture did not decrease intraocular pressure 20 min after acupuncture (P = 0.13) and 24 hours after acupuncture (P = 0.21). Nonetheless, acupuncture increased the effectiveness rate of treatment for glaucoma [OR = 45.00, 95% CI (9.73, 208.08)]. Compared with acupuncture, quantitative acupuncture manipulation increased the effectiveness rates of treatment for glaucoma [OR = 2.23, 95% CI (1.14, 4.36)].

CONCLUSIONAcupuncture therapy has potential to increase effectiveness rates of treatment for glaucoma. It lacks reliable evidence to prove that acupuncture decreases intraocular pressure. More trials with high quality are needed to estimate adverse effects and cost effectiveness of acupuncture therapy.

Acupuncture Therapy ; Glaucoma ; drug therapy ; therapy ; Humans ; Ophthalmic Solutions ; therapeutic use ; Randomized Controlled Trials as Topic ; Treatment Outcome

OBJECTIVETo ascertain extraction technology condition for extract and flavonoids from Chrysanthum morifoliwn.

METHODThe optimizing ultrasonic extraction condition on the basis of extractive yield and flavonoids were determined by orthogonal design.

RESULTThe order of factors which affected the flavonoid extraction was extraction times > ethanol concentration > ultrasonic time > solvent quantity.

CONCLUSIONThe optimum ultrasonic extractions are A2B3C3D3. Compared with traditional extraction, ultraction method is timesaving, simple to operate, stable and need not be heated.

Chrysanthemum ; chemistry ; Drugs, Chinese Herbal ; analysis ; Flavonoids ; analysis ; Flowers ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo examine the effect of RhoA/Rho kinase signal pathway on TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.

METHODSThe 4th generation of primary cultured human dermal fibroblasts were stimulated with TGF-beta1, (10 ng/ml). The expression of alpha-SMA was detected after treatment with TGF-beta1, for 0, 3, 6, and 24 h. The expression of alpha-SMA was also detected after treatment with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml). Then the human dermal fibroblasts (4th generation) were stimulated with TGF-beta1, (10 ng/ml) after being treated with the RhoA/Rho kinase signaling pathway inhibitor Y-27632 (10 umol/ml). The fibroblasts were treated with nothing as sham control, or with Y-27632 (10 umol/L) only as negative control group, or with TGF-beta1 (10 ng/ml) only as positive control group. The expression of alpha-SMA was detected in all the groups. Protein expression was analyzed with ANOVA statistical method.

RESULTSalpha-SMA expression in fibroblasts with 10 ng/ml TGF-beta1 stimulation for 0, 3, 6, 24 h was 1.0, 1.9 0.2, 2.1 +/- 0. 1, 3. 1 +/- 0.1, respectively. Alpha-SMA expression in 24 h group was significantly higher than that in other three groups (n = 4, P < 0.05). alpha-SMA expression in human dermal fibroblasts after stimulation with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml) was 1.0, 1.4 +/- 0.2, 3.2 + 0.1, 3.1 +/- 0.2, respectively. alpha-SMA expression in 10 ng/ ml group was significantly higher than that in 2 ng/ml group and control group (n = 4, P < 0.05). There was no statistical difference in alpha-SMA expression between 10 ng/ml group and 50 ng/ml group (n = 4, P > 0.05). With both Y-27632 (10 micromol/L) and TGF-beta1 stimulation, the cell phenotype differentiation was inhibited. Alpha-SMA expression in experimental group (1.2 +/- 0.2) was significantly reduced, when compared with that in positive control group (2.9 +/- 0.1) (n = 5, P < 0.05). There was no significant difference (n = 5, P > 0.05) in alpha-SMA expression between control group (1.0) and negative control group (1.1 +/- 0.1).

CONCLUSIONSRhoA/Rho kinase signaling pathway should be involved in TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.

Actins ; metabolism ; Adolescent ; Cell Differentiation ; Cells, Cultured ; Fibroblasts ; cytology ; Humans ; Male ; Signal Transduction ; Skin ; cytology ; Transforming Growth Factor beta1 ; pharmacology ; rho-Associated Kinases ; metabolism ; rhoA GTP-Binding Protein ; metabolism

The purpose of this work was to investigate the effects of niflumic acid (NFA), a chloride channel blocker, on the proliferation of human hepatoma cell line (HHCC). Cell proliferation was analyzed by cell count and MTT assay. Cell cycle analysis was carried out by flow cytometry. [Ca(2+)](i) was determined by laser scanning confocal system. It was found that NFA decreased significantly the cell number and the MTT optical density (OD) of HHCC cells, and that the OD value was reversed after washout of NFA. Compared with control, NFA blocked cell cycle progression in G(1) phase. Extracellular application of NFA (100 micromol/L) induced a rapid decrease in [Ca(2+)](i). These findings demonstrate that blockage of chloride channels by NFA induces growth arrest of HHCC in G(1) phase, which may be due to the inhibition of Ca(2+)/CaM-dependent signaling pathways.
Calcium ; metabolism ; Calmodulin ; metabolism ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chloride Channels ; antagonists & inhibitors ; Humans ; Liver Neoplasms ; pathology ; Niflumic Acid ; pharmacology

BACKGROUNDBone morphogenetic protein (BMP) is a member of the superfamily of transforming growth factor-beta. Recent studies show that it is an indispensable factor in hematopoiesis. To better characterize the effect of recombinant human BMP (rhBMP)-2 in hematopoiesis, we set out to determine whether rhBMP-2 could promote the proliferation of mesenchymal stem cells (MSCs) and increase the levels of hematopoietic cytokines in MSCs.

METHODS2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino) carbonyl)-2H-tetrazolium hydroxide (XTT), real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the effects of rhBMP-2 on the proliferation and hematopoietic cytokine levels of MSCs. In addition, MSCs marked with Hoechst33342 were transplanted into BALB/c mice by the intravenous route or intra-bone marrow transplantation, and cluster numbers were counted.

RESULTSThe XTT test revealed that rhBMP-2 significantly induced proliferation of MSCs in doses ranging from 10 ng/ml to 0.1 mg/ml in a dose-dependent manner. The experiments in vivo showed that there were more clusters of donor cells in bone marrow, spleen, liver and lung of the BMP group than those in the control group after both intra-bone marrow transplantation (P < 0.001, P < 0.001, P < 0.001, and P = 0.001, respectively) and intravenous transplantation (P < 0.001, P < 0.001, and P < 0.001 respectively). The results of real-time PCR and ELISA revealed that rhBMP-2 significantly increased mRNA expressions and protein levels of IL-6, IL-7, IL-11, G-CSF, M-CSF and SCF.

CONCLUSIONSThe treatment with rhBMP-2 promotes the proliferation of MSCs in vivo and in vitro and increases the levels of hematopoietic cytokines in MSCs, which may contribute to the improvement of hematopoietic function.

Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Granulocyte Colony-Stimulating Factor ; genetics ; Humans ; Interleukin-11 ; genetics ; Interleukin-6 ; genetics ; Interleukin-7 ; genetics ; Macrophage Colony-Stimulating Factor ; genetics ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Polymerase Chain Reaction ; Recombinant Proteins ; pharmacology ; Stem Cell Factor ; genetics ; Transforming Growth Factor beta ; pharmacology

BACKGROUNDIncreased proliferation of pulmonary vascular cells and muscularisation of pulmonary vessels are frequently observed in human smokers and in animals exposed to cigarette smoke. To elucidate the molecular mechanisms leading to these changes, we studied the in vitro effect of cigarette smoke extract (CSE) on proliferation of pulmonary artery smooth muscle cells (PASMCs) and activation of protein kinase C (PKC), an important kinase implicated in cell proliferation.

METHODSPASMCs cultured from 12 normal Wistar rats were studied in the following conditions: (1) PASMCs were exposed to different concentrations of CSE for 24 hours, then MTT colorimetric assay was used for detection of cell proliferation. Cell viability was assessed by trypan blue exclusion. (2) PASMCs were pre-incubated with phorbol 12-myristate 13-acetate (PMA) for 24 hours or Ro31-8220 for 30 minutes before exposure to 5% CSE for 24 hours. Cell proliferation was examined by MTT colorimetric assay, cell cycle analysis and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. (3) PASMCs were exposed to 5% CSE for 24 hours. Then PKC-alpha mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) and protein expression by Western blotting, while PKC-alpha translocation was observed by immunofluorescence staining and confocal microscopy. (4) PASMCs were transfected with specific antisense oligodeoxynucleotides against PKC-alpha 6 hours before exposure to 5% CSE for 24 hours. PKC-alpha protein expression and cell proliferation were detected by methods described previously.

RESULTS(1) Low concentration of CSE (5%) increased proliferation of PASMCs, whereas high concentrations (20%, 30%) were inhibitory as a result of cytotoxicity. (2) The value of absorbance (Value A), proliferation index (PI), S-phase cell fraction (SPF) and average optical density of PCNA staining in PASMCs from 5% CSE exposure group (0.306 +/- 0.033, 0.339 +/- 0.033, 0.175 +/- 0.021, 0.315 +/- 0.038, respectively) were significantly increased compared with those of control group (0.249 +/- 0.018, 0.177 +/- 0.055, 0.092 +/- 0.023, 0.187 +/- 0.022, respectively) (P < 0.05). PKC down-regulation by PMA pretreatment or PKC inhibition by Ro31-8220 pre-incubation abolished the effect of 5% CSE on PASMCs proliferation. (3) After exposure to 5% CSE for 24 hours, PKC-alpha mRNA and protein expression in PASMCs (1.054 +/- 0.078, 1.185 +/- 0.041, respectively) were much higher than in untreated cells (0.573 +/- 0.054, 0.671 +/- 0.055, respectively) (P < 0.01). Moreover, 5% CSE induced a translocation of PKC-alpha from cytoplasm toward the perinuclear area and into the nucleus. (4) Specific antisense oligodeoxynucleotides against PKC-alpha reduced 5% CSE-induced expression of PKC-alpha protein (0.713 +/- 0.047 vs 1.180 +/- 0.056), also abolished the effect of 5% CSE on PASMCs proliferation significantly.

CONCLUSIONSCSE can be cytotoxic at high concentrations. But at low concentrations, it makes a mitogenic effect on cultured PASMCs. PKC, especially its alpha isozyme, seems to play an important role in CSE-induced proliferation of PASMC.

Animals ; Cell Proliferation ; Cells, Cultured ; Indoles ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Protein Kinase C-alpha ; antagonists & inhibitors ; physiology ; Pulmonary Artery ; cytology ; Rats ; Rats, Wistar ; Smoke ; Tobacco