Studies confirm that gefitinib treatment of epidermal growth factor receptor(EGFR)muta-tions for patients with non-small cell lung cancer(NSCLC)has a clear effect,and with radiotherapy sensitiza-tion effect. Many studies both at home and abroad show that gefitinib combined with radiotherapy can signifi-cantly improve the survival times of patients,especially for the elderly patients or the patients with brain metas-tases,which has fewer adverse reactions and with higher life qualities. Therefore,gefitinib combined with radiotherapy will be an effective treatment for the patients with advanced NSCLC.

Objective To investigate the effect of intrathecal gastrodin on skin cancer pain in mice.Methods Thirty-two female Balb/c mice,aged 8 weeks,weighing 20-25 g,were randomly divided into 4 groups (n=8 each) using a random number table:sham operation group (group S),skin cancer pain group (group SCP),gastrodin group (group G),and artificial cerebrospinal fluid (ACSF) control group (group ASCF).Skin cancer pain was produced by injecting phosphate buffer solution 20 μl containing about 2 ×105 4T1 breast cancer cells into the plantar surface of the left hindpaw.At 14th day after inoculation of cancer cells,ASCF 5 μl was injected intrathecally in S and ACSF groups,and gastrodin 150 μg/kg (5 μl) was injected intrathecally in group G.Before inoculation,at 30 min before intrathecal injection,and at 15,30,60,90,120 and 150 min after intrathecal injection,the thermal paw withdrawal latency (TWL) was measured.The expression of acid-sensing ion channel (ASIC)-3 mRNA in the spinal dorsal horn was detected using the real-time reverse transcriptase polymerase chain reaction after the last measurement of the pain threshold.Results Compared with group S,the TWL was significantly decreased at each time point before and after intrathecal injection in SCP,ACSF and G groups,and the expression of ASIC-3 mRNA in the spinal dorsal horn was significantly down-regulated in group G (P＜0.05).Compared with group SCP,the TWL was significantly increased at each time point after intrathecal injection,and the expression of ASIC-3 mRNA in the spinal dorsal horn was significantly down-regulated in group G (P＜0.05),and no significant change was found in the parameters mentioned above in group ACSF (P＞0.05).Conclusion Intrathecal gastrodin can reduce skin cancer pain and down-regulate ASIC-3 expression in the spinal dorsal horn which is helpful in maintaining the analgesic effect in mice.

Objective: The quality standards for Zhixuan Granule (Rhizoma Alismatis, Rhizoma Atractylodis Macrocephalae, etc.) were studied. Methods: The TLC methods for identification of Rhizoma Alismatis、 Rhizoma Atractylodis Macrocephalae were established. A simple HPLC was established for the determination of 23-acetate alisol B. The mobile phase was acetonitrile-water(70∶30). UV detecting wavelength was at 208nm. Results: Rhizoma Alismatis and Rhizoma Atractylodis Macrocephala could be detected. 23-acetate alisol B showed a linear relationship at the concentration range of 99～1388.8ng, r=0.9999. The average recovery was 103.05% and RSD was 2.41%(n=6). Conclusion: This method is suitable for the quality control of Zhixuan Granule.

ObjectiveTo establish a novel rapid detection method based on PCR-single-strand-conformational polymorphism (PCR-SSCP) to determine mutation of streptomycin-resistance associated rpsL and rrs genes in isolates of Mycobacterium tuberculosis (MTB).MethodsStreptomycin-resistance of 112 MTB isolates was detected using the routine drug susceptibility test,and a special PCR-SSCP assay was established.The mutations of rpsL and rrs genes in streptomycin-resistant MTB isolates were detected by PCR-SSCP and PCR direct sequencing (PCR-DS) ; the results from two techniques were compared.Results All isolates had both rpsL and rrs genes.Fifty-two isolates (46.4％) were streptomycin susceptible,in which only 1 isolate showed abnormal PCR-SSCP fragments from rrs gene,and the specificity of PCR-SSCP was 98.1％ (51/52).Sixty isolates (53.6％) were streptomycin-resistant,in which 46 (76.6％) and 11 ( 18.3％ ) isolates presented the abnormal PCR-SSCP fragments of rpsL and rrs gene,respectively.One streptomycin-resistant isolate showed abnormal PCR-SSCP fragments from both rpsL and rrs genes.The sensitivity of PCR-SSCP was 93.3％ (56/60).ConclusionThe PCR-SSCP that established in this study is a specific and sensitive method for rapid detection of the streptomycin-resistance associated mutations in rpsL and rrs genes of MTB.

Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

Objective To detect the expression of Zbtb7 gene in Phorbol esters(PMA) induced differentiation of leukemia cell lines.Methods 50 nmol/L PMA was used to induce the differentiation of U937 and K562 cell lines.The expression of Zbtb7 was detected by real-time PCR in the cells after induction on day 0,1,3 and 5,respectively. Results The expression of Zbtb7 in leukemia cell lines was markedly enhanced after treated by PMA(P

Objective To detect the mRNA and protein expression of downstream quinine nicotinamide adenine dinucleotide (NADH) dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) induced by blood nuclearrelated factor E2 (Nrf2) in the peripheral blood of those exposed to arsenic in the endemic area of coal arsenic poisoning in Guizhou Province,and to discuss its role in the process of occurrence and development of liver injury due to coal arsenic poisoning.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey sites,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group based on physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.Peripheral blood samples were collected from all subjects.And mRNA expression of NQO1 and HO-1 were detected by RT-qPCR.Content of NQQ1 and HO-1 were detected by enzyme linked immunosorbent assay (ELISA).Results (①)Results of mRNA expression of NQ01 and HO-1:the relative expression level of mRNA of NQO1 and HO-1 in peripheral blood went up gradually as the degree of liver injury of population exposed to arsenic increased (F =5.548,10.961,all P ＜ 0.05);thereinto,the relative expression level of mRNA of NQO1 of mild hepatosis group (median:0.918) was higher than that of the control group (0.576,P ＜ 0.05),the relative expression level of mRNA of NQO1 of moderately severe hepatosis group (1.243) was higher than those of control group,non-patient group (0.653),non-apparent hepatosis group (0.636) and mild hepatosis group (all P ＜ 0.05);the relative expression level of mRNA of HO-1 of non-patient group (1.059),non-apparent hepatosis group (1.225),mild hepatosis group (1.553) and moderately severe hepatosis group (1.604) were higher than that of control group (0.767,all P ＜ 0.05);the relative expression level of mRNA of HO-1 of mild hepatosis group was higher than that of non-patient group (P ＜ 0.05);the relative expression level of mRNA of HO-1 of moderately severe hepatosis group was higher than those of non-patient group and non-apparent hepatosis group (all P ＜ 0.05).②)Results of protein expression of NQO-1 and HO-1:the level of protein expression of NQO1 and HO-1 in serum went up gradually as the degree of liver injury of population exposed to arsenic increased (F =19.685,17.725,all P ＜ 0.05).Thereinto,the protein expression of NQO1 and HO-1 of non-apparent hepatosis group,mild hepatosis group and moderately severe hepatosis group [NQO1:(6.272 ± 0.744),(6.336 ± 0.628),(6.714 ± 0.540) U/L;HO-1:(45.150 ± 4.813),(45.283 ± 5.049),(48.610 ± 5.365) U/L] were higher than those of control group and non-patient group [NQO1:(5.550 ± 0.730),(5.750 ± 0.427) U/L;HO-1:(39.856 ± 5.249),(42.375 ± 3.014) U/L,all P ＜ 0.05],the protein expression of NQO1 and HO-1 of moderately severe hepatosis group were higher than those of non-apparent hepatosis group and mild hepatosis group (all P ＜ 0.05).Conclusion The expression of mRNA and protein of NQO1 and HO-1 is closely related to the occurrence and development of liver injury due to arsenic exposure in coal arsenic poisoning areas.

Objective To investigate the effects of sodium arsenite (NaAsO2) on the expressions of Cyclin D1 and Cyclin dependent kinase 4 (CDK4) in human normal liver cells (L-02).Methods L-02 cells were exposed to different doses of NaAsO2 (0,50,100,150 μmol/L) for 24 h,flow cytometry was used to detect the cell cycle,real time quantitative PCR and Western blotting were used to detect the Cyclin D1,CDK4 mRNA and protein expression.Results Cell cycle detection:compared with the control group G0-G1 phase [(60.33 ± 0.40)％],except 50 μmol/L NaAsO2 group [(54.58 ± 0.40)％],the numbers of cells in 100 and 150 μmol/L NaAsO2 groups [(64.60 ± 0.62)％,(83.13 ± 0.25)％] were increased,the differences were statistically significant (all P ＜ 0.05);cell proportion of S phase in the control group,50,100 and 150 μmol/L NaAsO2 groups [(34.35 ± 0.30)％,(29.89 ± 0.32)％,(29.50 ± 0.44)％,(11.93 ± 0.12)％] were decreased,the differences were statistically significant (all P ＜ 0.05);cell proportion of G2-M phase was compared between the control group,50,100 and 150 μmol/L NaAsO2 groups [(5.32 ± 0.11)％,(15.54 ± 0.14)％,(5.90 ± 0.20)％,(4.93 ± 0.15)％],the difference was statistically significant (F =908.359,P ＜ 0.05).Cyclin D1 and CDK4 detection:the expressions of Cyclin D1 mRNA (0.48 ± 0.17,1.00 ± 0.31,1.00 ± 0.21,2.06 ± 0.53) and protein (0.65 ± 0.02,0.64 ± 0.05,0.71 ± 0.10,0.84 ± 0.05) were compared betwee the control group,50,100 and 150 μmol/L NaAsO2 groups,the differences were statistically significant (F =167.886,46.575,all P ＜ 0.05);the expressions of CDK4 mRNA (1.04 ± 0.19,1.00 ± 0.21,1.29 ± 0.22,2.31 ± 0.31) and protein (0.67 ± 0.04,0.74 ± 0.03,0.83 ± 0.07,0.94 ± 0.04) were compared betwee the control group,50,100 and 150 μ mol/L NaAsO2 groups,the differences were statistically significant (F =95.171,145.848,all P ＜ 0.05).Conclusions Treatment of L-02 cells with NaAsO2 has changed the expressions of Cyclin D1,CDK4 mRNA and protein,which leads to L-02 cell cycle arrested at G0-G1 phase,ultimately leads to cell damage.

AIM and METHODS: To investigate the expression of adhesion molecule ? 2 integrins (CD11a、CD11b) and L-selectin(CD62L )on Acute Lymophocyte Leukemia(ALL) cells and its Clinical Implications. Adhesion molecules CD11a、CD11b、 CD62L of 45 ALL patients and 25 health people were measured by flow-cytometric analysis. RESULTS:①CD11a and CD11b expression were lower on ALL cells than the normal hematopoietic cells. The rate of low expression was 100% for CD11b, 50% for CD11a,respectively. CD62L expression were higher on ALL cells than the normal hematopoietic cells.②The CD11a was lower expressed on B-ALL than T-ALL. CD62L was higher on T-ALL than B-ALL. ③ The expression of CD11a in the invasion group was much higher than that in the non-invasive group(P

[Objective] To compare the effect and safety between Tripterygium Wilfordii Film (TNF) and Compound Tripterygium Wilfordii Film (CTWF) for osteoarthritis of knees. The latter was mainly composed of Radix Folium seu Flos Tripterygii Wilfordii, Rhizoma Chuanxiong, Olibanum, Myrrha, etc. [Methods] Sixty cases of osteoarthritis of knees were randomized to two groups: group A (30 cases) treated with CTWF and group B (30 cases) with TWF. Both of the two groups were given oral use of herbal medicine on the basis of syndrome differentiation. Pain and stiffness of the knees and knee function were observed before and after treatment and their side reactions were also observed. [Results] Pain and stiffness of the knees and the impaired function were relieved in both groups ( P