Objective To detect the proliferation inhibition and apoptosis of HL-60 cell induced by APS-1 and APS-2 isolated from Polyporus sp. M05 and to investigate its mechanism. Methods The proliferation inhibition was detected by living cells count method,and chosed proper concentration.Flow cytometry with propidium iodide staining was used to detect cell apoptosis. Semi-quantitative RT-PCR was used to detect the expression of apoptosis related gene. Results APS-1 and APS-2 could significantly inhibit the proliferation of HL-60 cells on a time and dose dependent manner. Apoptosis ratio increased to 47. 9% and 26. 8% after HL-60 cells were exposed to APS-1 and APS-2 respectively for 48 h,and the differences had statistical significance (P <0.01). After being induced by APS-1,mRNA of Bax,Fas,Caspase-3 was upregulated. And after being induced by APS-2,mRNA of Bax,Caspase-3 was upregulated,while Fas mRNA did not change. Conclusion APS-1 and APS-2 can inhibit the proliferation and induce apoptosis of HL-60 cells. Mechanism of HL-60 cell apoptosis induced by APS-1 is related to both mitochondrial pathway and Fas signaling pathway,while apoptosis induced by APS-2 is only related to mitochondrial pathway.

Objective To investigate the effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA. Methods After establishing the in vitro differentiation model with HMBA on K562 cells, the MTT assay was used to detect the proliferation of K562 cells, the cell cycle profile was detected by flow cytometry, and the mRNA expression of TGF-β1, SMAD3, SMAD4 and EVI1 was measured by RT-PCR assay. Results HMBA could inhibit the proliferation and promote the differentiation of K562 cells obviously, which was time and concentration-dependent, and the 72 h corresponding IC50, was about 2 mmol/L. Within 72 h, flow cytometry assay indicated that the ration of G0-G1 phase cells was up-regulated, and the results of RT-PCR showed that relative mRNA expression of TGF-β1, SMAD3 and SMAD4 at mRNA level was increased gradually while that of EVI1 was decreased gradually. Conclusion HMBA can inhibit K562 cells proliferation through TGF-β1/SMAD signaling pathway.

OBJECTIVETo explore the anesthetic effect of preemptive analgesia of frequency acupoint electrical stimulation on painless-induced abortion as well as its effect on anesthetics dosage.

METHODSNinety cases of early pregnancy who selected painless-induced abortion were randomly divided into two groups, 45 cases in each group. Frequency acupoint electrical stimulation at Ciliao (BL 32) and Shenshu (BL 23), disperse-densewave, 2 Hz/100 Hz in frequency for 15 to 20 min, was applied in the group A, which was followed by intravenous anesthesia of propofol. The intravenous anesthesia of propofol was applied in the group B. The blood pressure (BP), heart rate (HR) and SpO2 before, during and after surgery, anesthetic effect and dosage, waking time and adverse events were observed in the two groups.

RESULTSThe BP and HR during and after the surgery in the group A were not statistically different from those before the surgery (all P > 0.05). The BP was reduced and HR was slowed down during the surgery in the group B, which was significantly different from those before the surgery as well as those in the group A (all P < 0.05). The dosage of propofol was (114. 3-+6. 1) mg in the group A. obviously less than (193.2 +/- 8.9) mg in the group B (P < 0.05). The waking time was (5.6 +/- 1.2) min in the group A, obviously less than (10.1 +/- 3.9) min in the group B (P < 0.05). As for anesthetic effect, the incidence of Grade I in the group A was more than the group B (P < 0.05). The adverse events, including nausea, vomiting and contractions pain in the group A were evidently less than those in the group B (all P < 0.05).

CONCLUSIONThe preemptive analgesia of frequency acupoint electrical stimulation could significantly improve anesthetic effect of painless-induced abortion, reduce dosage of anesthetics, shorten waking time of surgery and guarantee the safety of surgery.

Abortion, Induced ; Acupuncture Analgesia ; Acupuncture Points ; Adult ; Electric Stimulation ; Female ; Humans ; Pain Management ; Pregnancy ; Young Adult

OBJECTIVETo investigate the effect of Gli1 gene silencing by RNA interference (RNAi) on proliferation of K562 cells and its mechanisms.

METHODSThe small interference RNA (siRNA) was synthesized in vitro. K562 cells were transfected with Gli1 siRNA by the way of lipofection (lipofectamine 2000). Non-specific siRNA transfected cells were used as control. Transfection efficiencies of different siRNA concentrations were detected by flow cytometry and the best siRNA concentration was selected. The silencing effect of siRNA was demonstrated by real time PCR and Westem blot analysis. Cell proliferation was measured by MTT method, cell cycle by PI assay, c-myc and p21 mRNA level was detected by real time PCR analysis.

RESULTSTransfection efficiency of siRNA was increased in a dose-dependent manner when siRNA concentration was below 200 pmol, and the highest transfection efficiency reached (80.11 ± 5.63)%. Both the mRNA and protein level of Gli1 was down-regulated in Gli1 specific siRNA group, the mRNA level was (52.60 ± 3.57)% of that of control group after 24 h (t = 20.33, P < 0.01) and the protein level was (79.31 ± 5.58)% of that of control group after 48 h (t = 6.54, P < 0.01). The cell proliferation rate in Gli1 siRNA group was (94.41 ± 3.58)% (t = 2.40, P = 0.05) and (90.22 ± 3.34)% (t = 4.37, P < 0.01) of that of control group after 24 h and 48 h, respectively. G(2)/M cell cycle arrest was observed, the mRNA level of c-myc was down-regulated while p21 was up-regulated in Gli1 siRNA group after 24 h and 48 h (P < 0.05).

CONCLUSIONSTargeted silencing of Gli1 gene by RNAi inhibits the proliferation of K562 cells, which acts through the down-regulation of c-myc and up-regulation of p21 expression.

Cell Proliferation ; Gene Silencing ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transcription Factors ; genetics ; Transfection ; Zinc Finger Protein GLI1

Objective To explore the methods of nursing quality management in order to provide safety,high quality and satisfactory nursing for patients,Methods To establish one grid management model of nursing quality combined with " double three-level".Then,the report of security events,the implemention of nursing consultation,the incidence of adverse events,and patients' satisfaction were observed and compared before and after the management model.Results Mter implementing the grid management model of nursing quality,satisfaction average scores of hospitalized patients,emergency department patients,discharged patients were ( 98.22 ± 1.12 ),( 91.97 ± 1.67 ),(97.16 ± 2.41 ) respectively,and before it was implemented the average scores respectively were ( 91.59 ± 1.67 ),( 87.92 ± 1.98 ),( 87.89 ± 1.81 ),and the differences were statistically significant ( t =3.57,2.69,4.72,respectively ; P ＜ 0.05 ).There was a significant change in the report of security events,implemention of nursing consultation and checking.The incidence of administration errors,falling down and pressure sores were significantly reduced.Conclusions Construction of the grid management system of nursing quality can ensure the implemention of nursing core institution,improve risk awareness of nurses,improve the nursing quality and increase patients' satisfaction.

The purpose of this study is to investigate the enantioselectivity of norgestrel (NG) transdermal permeation and the potential influence of linalool and lipids on the enantioselectivity. In vitro skin permeation studies of NG across the excised rat skins were performed with Valia-Chien diffusion cells, and the permeation samples were analyzed by enantioselective HPLC. The possible enantioselective permeation of NG across intact rat back skin and lipids extracted rat back skin and the influence of linalool were evaluated. The skin permeation rate of dl-NG was two times higher than that of l-NG when donor solutions (EtOH/H2O 2 : 8, v/v) containing l-NG or dl-NG. It may be mainly attributed to the solubility discrepancy between enantiomer and racemate. The enantioselective permeation of dl-NG across intact rat skin was observed when the donor solutions containing dl-linalool. The permeation flux of l-NG was 22% higher than that of d-NG. But interestingly, the enantioselective permeation of dl-NG disappeared under the same experimental condition except that the lipid extracted rat skin was used. Attenuated total reflection-fourier transform infrared spectroscopy analysis of stratum corneum showed that the wave number for asymmetric CH2 stretching vibrations of lipids treated with dl-linalool was greater than that of the control. The results indicated that the enantioselective permeation of NG may be contributed by the interaction between dl-linalool and lipids. More than half of lipids were composed of ceramides. The stereospecific interaction maybe existed among chiral enhancer (linalool), lipids (ceramides) and/or chiral drugs (NG).
Administration, Cutaneous ; Animals ; Lipids ; pharmacology ; Monoterpenes ; pharmacology ; Norgestrel ; pharmacokinetics ; Rats ; Skin Absorption ; drug effects ; Spectroscopy, Fourier Transform Infrared ; Stereoisomerism

Objective To evaluate the positive effects of cisplatin on sensitivity of human glioma U251 to tumor necrosis factor-related apoptosis inducing ligand and to investigate the potential mechanism. Methods The expression of green fluorescent protein (GFP) in U251 which was transfected with pAdxsi-GFP-TRAIL was observed by inverted fluorescent microscope ×400) and to ascertain the MOI. The proliferation inhibition was studied by MTT method. Morphological change was detected through inverted florescent microscope and the Hoechst33342 staining assay was used to verify whether cell apoptosis could be induced or not. The cell apoptosis was also analyzed by flow cvtometry with propidium iodide staining. Semi-quantitative RT-PCR was introduced to detect the mRNA expression of apoptosis related gene.Results The expression of TRAIL mRN A was significantly upregulated after transfection. Compared with treatment group of cisplatin and TRAIL alone, the proliferation of U2S1 was significantly inhibited in the cisplatin sensitizing TRAIL group (P < 0.05 ). Nuclear shrinkage and pyknosis fragmentation were observed by Hoechst 33342 staining assay; Apoptotic peak was detected from the results of flow cvtometry and there were significant differences between the sensitizing group and the other two groups ( P < 0.05 ) ; Moreover, the relatively high expression of TRAIL, DR5, caspaseS and down - regulated survivin genes were also observed. There was no significant changes in DR4 expression. Conclusion Cisplatin could extremely enhance the sensitivity of U251 cells to TRAIL And the potential mechanism may related to the increase of TRAIL, DRS, caspaseS genes while the reduction of surivivin gene.

Abstract: Objective　To investigate the clinical characteristics and incidence of Brucella encephalitis and meningitis in children. Methods We report the clinical data of a child with Brucella melitensis meningitis in children, and summarize the incidence, diagnosis methods and treatment of Brucella encephalitis or meningitis in children, taking into account the relevant domestic and foreign literature from January 2014 to December 2020. Results　A 4-year-old girl was admitted to the hospital with status epilepticus on March 15, 2021 because of interrupted right limb numbness for 16 hours and convulsions for 2 hours. She had 2 non-febrile convulsions three months before admission and was diagnosed with epilepsy. This incident was acute, accompanied by low fever, with epilepsy as the main manifestation. Cerebrospinal fluid test suggested central nervous system infection, but the nature of infection could not be determined by routine and biochemistry of cerebrospinal fluid.The cerebrospinal fluid next generation sequencing confirmed that the pathogen of the infection was B. melitensis, which was further verified by the peripheral blood antibody test. After effective antibiotics combined with a full course of treatment, the patient recovered after six months of treatment. A total of 60 articles were retrieved in the database, including 29 in Chinese. During this period, a total of 7 cases of brucellosis in children with nervous system involvement were reported, one of which was a case report, and the other 6 cases were mentioned in the comprehensive analysis of children with brucellosis. Conclusions　Brucella encephalitis or meningitis in children has a low incidence and various clinical features, which are easy to be misdiagnosed or missed.

Objective To survey the sleep quality status of patients with ankylosing spondylitis,and explore the integrative methods of care to improve the sleep quality of them with sleep disorders.Methods Using the Pittsburgh Sleep Quality Index (PSQI) to evaluate the sleep quality level of 36 patients with ankylosing spondylitis.Then,the combined nursing of traditional Chinese and Western medicine was implemented.When they discharge,their sleep quality was evaluated again.Results When they discharge,the total sleep hours,sleeping habits and sleep quality of 32 cases were significantly improved,and their life habits,mental state and treatment compliance were improved in some degree,and had a good quality of sleep by PSQI evaluation (PSQI ＜ 7).Only 4 cases had different degree of sleep disorder.The PSQI score of patients was significantly decreased after and before admission [(5.46 ± 1.45) vs (10.35 ± 3.22) ;t =8.3 1,P =0.000].Conclusions The humanized combined nursing interventions of traditional Chinese and Western medicine can improve sleep quality of patients with ankylosing spondylitis significantly,and improve the patient's life and survival quality.

The study was aimed to investigate the mobilization effect of medium dose of granulocyte colony stimulating factor (G-CSF) in allogeneic peripheral stem cell transplantation and changes of T lymphocyte subgroup in PBMNC before and after mobilization. G-CSF was administered at 600 microg/d (i.e. 300 microg i.v. twice a day) for successive 5 days to 31 matched sibling or unrelated donors for the mobilization. Stem cells were harvested on the fourth day. FACS was used to analyze the NC, MNC and T lymphocyte subgroups. The results showed that the number of NC, MNC, CD34(+) cells and CFU-GM in dose of 600microg/d significantly increased (P < 0.05), compared with 300 microg/d; the time for hematological reconstruction was significantly shortened (P < 0.05); the ratio of adverse effects was not obviously increased (P > 0.05) and the median percentage of CD3(+) lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization, while the ratio of CD4(+)/CD8(+) did not significantly changed. It is concluded that the administration of G-CSF 600 microg/d in allo-PBSCT has a good effect in the mobilization of PBSC with minor side effects, which can markedly promote hematopoietic reconstitution after transplantation. The relative amount of CD3(+) lymphocytes significantly decreased and the ratio of CD4(+)/CD8(+) remained unchanged, which may lead to alleviation of a GVHD after PBSCT.
Adolescent ; Adult ; Antigens, CD34 ; analysis ; Blood Donors ; Female ; Flow Cytometry ; Graft vs Host Disease ; prevention & control ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; methods ; Recombinant Proteins ; T-Lymphocyte Subsets ; cytology ; drug effects ; immunology ; Time Factors