Objective To detect the effect of osthole on proliferation and apoptosis of HL-60 cells and its molecular mechanism.Methods HL-60 cells proliferation was measured through the CCK8 assay method.The cell morphological changes were observed by Hoechst33342 staining after 8 h of drug effect.Induction of apoptosis was determined by flow cytometry and fluorescent microscopy.Expressions of Bcl-2 and Bax mRNA were evaluated by RT-PCR,and the expressions of cleaved caspase-3,caspase-8,caspase-9,Fas and FasL were evaluated by using western bolt assay.Results Osthole could inhibit the proliferation of HL-60 cells,the maximum inhibiting rate was (90.7 ±4.5)％,F =138.46,P =0.000; the apoptosis rate was 33.6％,F =27.75,P =0.006.The changes of apoptosis of cells and nucleus were shown in cell morphological observation.Osthole affected the decrease of the mRNA levels of Bcl-2 and the increase of the Bax mRNA levels via a dosedependent manner(F =210.12,P =0.000).Western blotting demonstrated that osthole could lead to the increase of the expression levels of cleaved caspase-3,caspase-8,caspase-9,Fas and FasL in the HL-60 cell line via a time-dependent manner.Conclusion Data suggests that osthole inhibits proliferation and induces apoptosis of HL-60 cells through mitochondria-dependent pathway and death-receptor pathway.

Objective To explore the correlation between the strength of trunk flexors and extensors obtained by isometric and isokinetic concentric tests. Methods The strength of trunk extensors and flexors were measured by isometric and isokinetic concentric tests during trunk flexion and extension in 50 healthy subjects, and the peak torque of flexion and extension (FPT and EPT) and the ratios of FPT/EPT were analyzed with correlation and regression analysis. Results The FPT and EPT of trunk flexors and extensors were positively correlated (r_F=0.827,P0.05, respectively), the regression equations of FPT and EPT were y^=8.158+1.097X and y^=-12.375+0.781X, respectively. Conclusion The isometric test of the trunk extensors and flexors could assess accurately the changes of the trunk muscle strength without aggravating pain and diseases. It is more reasonable to evaluate the stability of trunk with the FPT/EPT ratio at neutral position.

BACKGROUND:With the development of modern pathological techniques, the misdiagnosis rate has been reduced remarkably, but special stains are still the most important method for pathological diagnosis. OBJECTIVE:To compare the advantages and disadvantages of different special stains used for observing the structure of human lumbar facet joints. METHODS:The specimens of facet joint cartilage at L4/5 level were collected from patients undergoing lumbar surgery, and then stained with hematoxylin-eosin, safranin O, toluidine blue, Masson, and saranin-O-fast green for structure observation. RESULTS AND CONCLUSION:The structure of the articular cartilage could be observed clearly through hematoxylin-eosin, toluidine blue, and saranin-O-fast green staining. The cartilage surface, tidemark, and subchondral bone were shown by the hematoxylin-eosin staining, with the presence of violet chondrocyte nuclei. Safranin-O-fast green staining showed the four layers of the cartilage clearly, including the shallow layer (cartilage surface), middle layer (spherical cells arranged in disorder), columnar cell layer (large and multinucleated chondrocytes arranged neatly), tidemark, subchondral bone layer; and the cartilage matrix was reddish uniformly, the subchondral bone was green, and the cartilage and bone tissue showed a striking contrast. The cartilage structure was unclear in toluidine blue staining, with clear nuclei and almost no coloring cytoplasm, but the matrix appeared with slight purplish blue. Safranin O staining showed that the cartilage was red, which had no obvious boundary with the cartilage matrix, and chondrocytes were stained lightly. Masson staining showed clear collagen fibers, but the structures of the cartilage and subchondral were obscure. To conclude, safranin-O-fast green staining can achieve the best results, followed by hematoxylin-eosin staining and Masson staining in turn.

Objective To detect genetic polymorphism of myeloperoxidase (MPO) gene and catalase (CAT) gene and their activities, and to analyze their relationship with arsenic poisoning caused by coal-burning. Methods One hundred and thirty arsenic poisoning patients were chosen as case group in Jiaole Village, Xingren County, Guizhou Province(an endemic area). One hundred and forty healthy residents living in 13 km away were chosen as control group. Their blood was collected. Polymerase chain reaction-restriction fragment length polymorphism technique(PCR-RFLP) was used to detect polymorphism of MPO-463G/A and CAT-262C/T. Ultraviolet spectmphotometer method was used to detect myeloperoxidase activity. Chromatometry method was used to detect catalase activity. Results The genotype frequency of MPO-463G/A at GG, GA, AA site was 47.24%(60/127), 44.09%(56/127),8.67% (11/127) in case group and 42.34% (58/137),48.17% (66/137)1,9.49% (13/137) in control group, respectively. The difference between the two groups was not significant(χ2 = 0.642, P > 0.05). The genotype frequency of CAT-262C/T, at CC, CT, TT site was 65.60%(82/125),28.80%(36/125),5.60%(7/125) in case group and 76.51%(101/132), 18.94% (25/132) ,4.55% (6/132) in control group, respectively, without significant difference (χ2 =3.845, P>0.05). The relationship between polymorphism of MPO-463G/A and CAT-262C/T and the risk of arsenic poisoning was not found in this study(ORadj= 1.36, 95%CI: 0.74-2.50 for MPO; ORadj=1.35, 95%CI: 0.69-2.63 for CAT). The activities of MPO and CAT were (25.30±8.70)U/L and (2.80± 1.09)×103 U/L in case group, while (22.76±7.59)U/L and (3.90±1.01)×103U/L in control group with a significant difference(F=0.760 for MPO, F=0.855 for CAT, all P < 0.05). The genotype of MPO-463G/A and CAT-262C/T was not found to have relationship with the activities of MPO, CAT(F=1.312,2.822 for MPO; F= 0.151,0.036 for CAT, P>0.05). Conclusions Genetic polymorphism of MPO-463G/A and CAT-262C/T is not found to have relationship with arsenic poisoning. Arsenic can lead to the change of MPO and CAT activity, which, however, may not be affected by MPO-463G/A and CAT-262C/T polymorphism.

Objective To explore the influence of coal-arsenic exposure on human T cells proliferation and its mechanism.Methods Blood samples colleoted from individuals which lived in arsenism area of coal-burning type and non-arsenism area in Guizhou Province were divided into exposed group(17),mild(35),moderate(38) and severe arsenism group(19)and control group(35)according to Diagnosis Smndard for Endemic Arsenism (WS/T 211-2001).T cell stimulation index wag determined by methyl thiazolyl tetrazolium(MTT)colorimetric method.The intracellular Ca2+ exponential(IECa2+)in peripheral blood mononuclear cell(PBMC)was analyzed by Fho-3/AM dye and flow cytometry.DNA binding activity of actively T cells nuclear factor(NF-AT)in PBMC was evaluated by electrophoretie mobility shift assay(EMSA).Results Concanavalin A(ConA)stimulation decreased the T cells stimulation indexes in exposed group,mild,moderate and severe arsenism groups(1.315±0.962, 1.611±1.224,1.114±0.545,1.289±0.875)compared with control group(2.322±1.241),all the differences being statistically significant(P＜0.01).After stimulated by anti-CD3 monoclonal antibody(McAb),the T cells stimulation index in exposed group,mild,moderate and severe arsenism group(0.997±0.177,1.103±0.291,1.007±0.221, 0.957±0.205) were lower than that of control group(1.842±0.429,P ＜ 0.01 ). IECa2+ of PBMC after treated by anti-CD3 McAb in mild,moderate and severe arsenism group( 110.130±49.637,92.429±31.191,77.640± 35.372) were lower compared with control group(145.986±59.450,P ＜0.01 ). Moreover,IECa2+ in moderat and severe arsenism group were lower than exposed group(121.337±46.410,P ＜ 0.05). DNA binding activity of PBMC NF-AT in mild,moderate and severe arsenism group(1.354±0.446,1.290±0.291,1.159±0.411 ) were lowered than that of control group(1.722±0.291,P ＜ 0.01) and exposed group(1.611±0.294,P ＜ 0.05). Conclusions The coal-arsenic exposure can reduce the human T cells stimulation indexes,IECa2+ in PBMC and the DNA binding activity of NF-AT. It suggest that arsenic may suppress the proliferation ability of human T cells,which may be partly related to the influence of arsenic on T cell receptor(TCR)/CD3 signal transduetion pathway.

Objective To investigate the activation of T lymphocytes in human peripheral blood and the signaling molecules in protein kinase C/nuclear factor KB(PKC/NF-κB) pathway expressivity or activity changes in human peripheral blood mononuclear cells(PBMCs) exposed to coal-arsenic,to explore the role of PKC/NF-κB signal pathway in activation of T cells in human exposed to coal-arsenic. Methods Blood samples were collected from individuals who lived in arsenism area of coal-burning in Guizhou province, and were divided into asymptomatically exposed group (12),mild arsenism group (33),moderate arsenism group (34) and severe arscnism group (15) according to Diagnosis Standard for Endemic Arsenism (WS/T 211-2001). The individuals who lived in non-arsenism area were control group(27). The ratio of activated T ceils was analyzed by flow cytometry. DNA binding activity of NF-κB in PBMCs was evaluated by electrophoretic mobility shift assay(EMSA). The expression of PKCθ and phospho-PKCθ(pPKCθ) in PBMCs were detected with western blotting analysis. Results The ratio of activating T cells in asymptomatically exposed group[(21.76±15.31)%],mild arsenism group[(18.41±11.36)%],moderate arsenism group[(17.78±11.93)%]and severe arsenism group[(18.79±13.38)%]were all higher than that of control group[(3.19±2.12)%],the difference among all groups being statistically significant(F = 7.893,P < 0.05). DNA binding activity of PBMCs NF-κB in asymptomatically exposed group,mild arsenism group,moderate arsenism group and severe arsenism group(1.49±0.24,1.58±0.30,1.57±0.34,1.51±0.16) were higher than that of the control group(1.30±0.17),the difference being statistically sign/ficant(P < 0.05 or < 0.01). The expression of PBMCs pPKCθ in mild arsenism group,moderate arsenism group and severe arsenism group(0.64± 0.14,0.64±0.27,0.62±0.12) were all lower than that of the control group(0.93±0.20),the difference being statistically significant(P < 0.05). There were significant negative correlations between the expression of pPKCθ and the activity of NF-κB(r =-0.565,P < 0.01). There were significant positive correlations between the activity of NF-κB and the ratio of activating T cells(r = 0.546,P < 0.01). Conclusion Coal-arsenic enhances the DNA binding activity of NF-κB,reduces the expression of PBMCs pPKCθ in human PBMCs and up-regulates the activity of T cells. It suggests that the PKC/NF-κB signal might be one of transduction pathway via activating of T cells by coal-arsenic.

Objective: To investigate the breakthrough points and methods of pharmaceutical care performed by clinical pharmacists for chemotherapy-induced Ⅳ degree myelosuppression.Methods: One advanced lung adenocarcinoma patient suffering from chemotherapy-induced Ⅳ degree myolosuppression was selected as the example, clinical pharmacists provided suggestions on the chemotherapy regimen, assisted physicians in making reasonable treatment plan and implemented comprehensive pharmaceutical care for the patient.Results: Clinical pharmacists played a positive role in the clinical treatment through the comprehensive pharmaceutical care in oncology department.Conclusion: With the professional knowledge of pharmacy, clinical pharmacists can improve the level of clinical rational drug use.

Hepatitis B virus (HBV) infection could lead to different clinical presentations and disease progresses, including acute and chronic hepatitis B, liver cirrhosis, hepatocellular carcinoma, and occult HBV infection, in which the interaction between virus and host plays an important role. Because HBV reverse transcriptase is lack of correction function, HBV is prone to generate mutations under the pressure of host immune response and antiviral drug treatment. Some mutations in the HBV S gene-encoding region can significantly attenuate antibody immune response against HBV and therefore affect clinical presentations and disease progression. Such mutations are termed immune escape-related mutations. In this paper, we mainly review the structure and functional characteristics of HBV S gene, the causes and forms of the immune escape-related mutation, its influence on clinical presentations and antiviral treatment response, as well as clinical detection methods.

OBJECTIVETo compare the clinical efficacy differences between fire needling technique of filiform needle at high stress points and regular acupuncture on chrondromalacia patellae so as to provide the better therapy for the treatment of this disease.

METHODSSixty cases of chrondromalacia patellae were randomized into a fire needling group (28 cases) and a routine acupuncture group (32 cases). In the fire needling group, 5 to 6 high stress points were localized according to the symptoms, palpation and imaging condition and were stimulated with fire needling technique of filiform needle. The treatment was given once every two days, 5 treatments made one session. In the routine acupuncture group, the regular acupuncture was applied at Dubi (ST 35), Xiguan (LR 7), Yanglingquan (GB 34), Yinlingquan (SP 9), Zusanli (ST 36), etc. The treatment was given once every day, 5 treatments made one session. Lysholm score, VSA score, patella title angle (PTA) and lateral patella angle (LPA) of the affected knees before and after treatment, as well as the clinical efficacy after treatment were observed in the two groups.

RESULTSAfter treatment, Lysholm score, VSA score, PTA and LPA were all improved apparently in the two groups (all P < 0.01). After the treatments, the improvements in Lysholm score, VSA score, PTA and LPA in the fire needling group were more obvious than those in the routine acupuncture group (all P < 0.05). The effective rate was 92.9% (26/28) in the fire needling group, better than 87.5% (28/32) in the routine acupuncture group (P < 0.01).

CONCLUSIONThe fire needling technique of filiform needle at the high stress points relieves the clinical symptoms of chrondromalacia patellae and recovers the biodynamical structure of patellae.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adult ; Chondromalacia Patellae ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To analyze the mutation characteristics of immune escape-associated mutations in the main hydrophilic region (MHR) of HBsAg in patients with drug-resistant mutations in HBV RT. Methods The clinical data were analyzed from 6917 patients with C-genotype HBV infection attending to the hospital from July 2007 to August 2017. And these patients were treated with nucleoside/nucleotide analogs (NAs) and received drug resistance tests. The detection rate of drugresistant mutations in HBV RT was determined and the mutation characteristics of MHR immune escape-related mutations in HBsAg between drug-resistant mutations of HBV RT group and wild-type of HBV RT group were compared. Results Classical drug-resistant mutations in HBV RT occurred in 2585 chronic hepatitis B (CHB) patients treated with antiviral therapy, and the overall detection rate was 37.37% (2585/6917). The overall mutation rate of MHR in the drug-resistant mutations group of HBV RT was significantly higher than that in the wild-type group (30.00% vs. 17.13%, P<0.05). It was found that the detection rates of sQ101K/R/H, sS114T/A/L, sT/I126S/N/A, sG130N/R/K/A, sM133T/I, sS143T/L, sA159V/G, sE164D/G in HBV RT drug-resistant mutations group were higher than those in RT wild-type group (P<0.05). Conclusions Patients with HBV RT resistant mutations have a higher detection rate of MHR immune escape mutations in HBsAg, which suggested that MHR immune escape related mutations were closely related to HBV RT drug-resistant mutations.