DNMT1 protein promotes retinoblastoma proliferation by silencing MEG3 gene / 南方医科大学学报

OBJECTIVE@#To investigate whether DNMT1 protein induces retinoblastoma proliferation by silencing MEG3 gene.@*METHODS@#Two retinoblastoma cell lines (HXO-RB44 and SO-RB50) and a normal human retinal pigment epithelial (RPE) cell line were transfected with the plasmid pcDNA-DNMT1 or si-DNMT1 for up-regulating or interference of DNMT1 expression, and with pcDNA-MEG3 or si-MEG3 for up-regulating or interference of MEG3 expression. Western blotting was used to detect the changes in the expression of DNMT1 protein in the transfected cells, and CCK-8 and EdU assays were used to detect the changes in cell proliferation. Real-time quantitative PCR (qRT-PCR) was performed to detect MEG3 expression in SO-RB50 and HXO-RB44 cells after transfection, and the methylation level of MEG3 gene promoter after interference of DNMT1 expression was detected using methylation-specific PCR.@*RESULTS@#SO-RB50 and HXO-RB44 cells showed significantly increased expression of DNMT1 protein as compared with normal RPE cells ( < 0.05). In HXO-RB44 cells, transfection with pcDNADNMT1 resulted in significantly increased expression of DNMT1 protein, enhanced cell proliferation ability, and significantly reduced expression of MEG3 ( < 0.05). In SO-RB50 cells, transfection with si-DNMT1 significantly reduced the expression of DNMT1 protein, suppressed the cell proliferation, and increased MEG3 expression ( < 0.05). Interference of DNMT1 significantly reduced the methylation level of MEG3 gene promoter. After reversing the regulatory effect of DNMT1 on MEG3 gene, DNMT1 protein showed significantly weakened ability to regulate retinoblastoma cell proliferation ( < 0.05).@*CONCLUSIONS@#In retinoblastoma cells, the up-regulation of DNMT1 protein induces promoter methylation and inactivation of MEG3 gene and eventually leads to abnormal cell proliferation.

Simultaneous detection of 3 kinds of polycyclic aromatic hydrocarbons in the air of workplace by high performance liquid chromatography / 中国职业医学

OBJECTIVE: To establish a method for simultaneous detection of 3 kinds of polycyclic aromatic hydrocarbons(PAHs) including phenanthrene,anthracene and 3,4-benzo( a) pyrene in workplace air by high performance liquid chromatography( HPLC). METHODS: The phenanthrene,anthracene and 3,4-benzo( a) pyrene were collected in the air of the workplace using glass fiber filter paper. The membrane sample was added with 5. 00 m L of acetone,and the samples were extracted by ultrasonic wave for 70 mintues and eluted with acetonitrile-water gradient. Liquid chromatography-UV detector with tandem fluorescence detector was used for determination. RESULTS: The good linearity rang of phenanthrene,anthracene and 3,4-benzo( a) pyrene was 0. 050-1. 000 mg/L with the correlation coefficients ≥0. 999 5; the detection limits were 0. 010,0. 007 and 0. 008 mg/L,and the minimum detectable concentrations were 0. 130,0. 090 and 0. 110μg/m3 respectively( air collection of 375 L). The desorption efficiencies were 86. 10%-99. 20% of phenanthrene,88. 60%-96. 40% of anthracene,and 84. 80%-99. 60% of 3,4-benzo( a) pyrene,respectively. The within-run relative standard deviation( RSD) were 1. 29%-3. 25%,1. 90%-3. 61% and 1. 30%-4. 82% respectively,the between-run RSD were 2. 41%-4. 07%,2. 02%-5. 12% and 2. 08%-4. 77% respectively. The standard recovery rates were 95. 00%-106. 14% of phenanthrene,93. 14%-106. 50% of anthracene, and 92. 86%-105. 50% of 3,4-benzo( a) pyrene,respectively. The samples could be stored at 4 ℃ for 7 days. CONCLUSION: This method is simple,accurate and reliable for the simultaneous detection of phenanthrene,anthracene and 3,4-benzo( a) pyrene in the air of workplace.

Clinicopathological characteristics of infectious granulomas: an analysis of 39 cases / 中华全科医师杂志

Objective To analyze the clinocopathological characteristics of infectious granulomas.Methods The clinical features,histopathological manifestations of 39 patients with infectious granulomas were analyzed retrospectively.Results Among 39 cases of infectious granulomas,there were 15 males and 24 females,and 17 cases of fungal granuloma and 22 cases of tuberculous granuloma.There was no statistically significant difference in gender and age between fungal granulomas and tuberculous granulomas.The mean course of tuberculous granuloma aud tuberculous granuloma was (0.88 ± 0.67) years and (5.54 ± 3.49) years,respectively (t =4.51,P =0.00);there was no significant difference in mean age of onset in fungal granuloma patients and tuberculous granuloma patients [(54.6 ± 19.6) vs.(47.6 ± 18.1) years,P ＞0.05)].There were 4 and 18 cases of fungal and tuberculous granulomatosis at the face,and 13 and 3 cases at the extremities (all P =0.00);the lesions occurred in the trunk in one case of tuberculous granuloma.The clinical manifestations of fungal and tuberculous granulomas as plaques/nodules were in 14 cases aud 22 cases (P =0.08);as ulcers and pus exudates were in 10 and 2 cases,respectively (P =0.00).The histopathological features showed epidermal hyperplasia in 12 and 4 cases,infiltrative patterns in 4 and 21 cases,infiltration of neutrophils in 14 and 3 cases,infiltration of plasma cells in 15 and 5 cases,infiltration of eosinophils in 10 and 0 cases,necrosis in 1 and 10 cases in fungal granulomas and tuberculous granulomas,respectively (P =0.00,0.00,0.00,0.00,0.00,0.01).Conclusion Fungal granuloma and tuberculous granuloma are different in the lesion sites,clinical manifestations and histopathological features.

Effect of narrow-band ultraviolet B radiation on the autophagy of cultured human melanocytes in vitro / 中华皮肤科杂志

Objective To evaluate the effect of narrow-band ultraviolet (NB-UVB) radiation on the autophagy of cultured human melanocytes in vitro,and to explore possible mechanisms underlying the treatment of vitiligo by NB-UVB.Methods In vitro cultured human melanocytes were divided into 4 groups to be irradiated with NB-UVB at different irradiation doses of 0 (control group),50,100 and 200 mJ/cm2 (50-,100-and 200-mJ/cm2 NB-UVB groups) respectively.After 24-hour treatment,the cells were collected,and monodansylcadaverin (MDC) staining was conducted to detect changes of autophagosomes in melanocytes.Western blot analysis was performed to determine the protein expression of autophagy signals including phosphorylated AMP-activated protein kinase (p-AMPK),phosphorylated mammalian target of rapamycin (p-mTOR),microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ (LC3 Ⅱ/Ⅰ) and P62,and transmission electron microscopy to observe ultrastructural changes of autophagosomes and melanosomes in the melanocytes.Statistical analysis was done by using one-way analysis of variance (ANOVA) for the comparison of Western blot results,and by Kruskal-Wallis H test for the comparison of the number of melanosomes,autophagosomes and autolysosomes.Results MDC staining showed that the percentages of autophagosome-positive melanocytes were significantly higher in the 100-,200-mJ/cm2 NB-UVB groups (38.08％ ± 4.10％,40.23％ ± 1.45％,respectively) than in the control group (21.83％ ± 3.50％,both P ＜ 0.05) and 50 mJ/cm2 NB-UVB group (23.66％ ± 4.12％,both P ＜ 0.05).As Western blot analysis revealed,the 100-,200-mJ/cm2 NB-UVB groups showed significantly increased expression of p-AMPK and LC3 Ⅱ/Ⅰ,but significantly decreased expression of p-mTOR and P62 compared with the control group (all P ＜ 0.05).Transmission electron microscopy showed that the number of autophagosomes and autolysosomes was significantly higher in the 100-,200-mJ/cm2 NB-UVB groups (5.12 ± 1.13,5.25 ± 1.04) than in the control group (1.88 ± 1.18,both P ＜ 0.05).Meanwhile,the number of melanosomes was significantly higher in the 50-,100-and 200-mJ/cm2 NB-UVB groups (39.12 ± 9.42,57.38 ± 7.11,59.75 ± 15.15,all P ＜ 0.05) than in the control group (18.50 ± 4.18,all P ＜ 0.05).Conclusion NB-UVB radiation can not only promote the formation of melanosomes,but also activate the autophagy signal pathways in the melanocytes and promote the formation of autophagosomes and autolysosomes,which may be one of the mechanisms underlying the treatment of vitiligo by NB-UVB.

Clinical analysis of 38 cases of steroid-induced diabetes mellitus due to glucocorticoid treatment / 中华皮肤科杂志

Objective To investigate risk factors for and clinical features of steroid-induced diabetes mellitus due to glucocorticoid treatment.Methods Clinical data were collected from 798 patients who received systemic glucocorticoid treatment in Department of Dermatology of Hangzhou Third People's Hospital from 2013 to 2016,and analyzed retrospectively.Logistic regression analysis was performed to analyze the factors influencing the occurrence of steroid-induced diabetes mellitus (SDM),repeatedmeasures analysis of variance to compare peripheral blood glucose levels of patients with SDM after breakfast,lunch and dinner,and t test to compare the levels of fasting blood glucose and glycosylated hemoglobin (HbA1 c) between patients with SDM and those with type 2 diabetes mellitus.Results Of the 798 patients,38 developed SDM due to glucocorticoid treatment.The average age was significantly older in the patients with SDM ([66.86 ± 13.30] years,n =38) than in those without SDM ([39.95 ± 17.01] years,n =760;t =8.86,P ＜ 0.01),but there was no significant difference in the gender ratio between the patients with and thhose without SDM (x2 =1.61,P =0.20).The prevalence of fatty liver,hyperlipidemia,hypertension,abnormal liver function and family history of diabetes mellitus was significantly higher in the patients with SDM than in those without SDM (x2 =12.25,19.25,32.69,21.47,16.70 respectively,all P ＜0.01).Logistic regression analysis showed that age,fatty liver,hyperlipidemia,hypertension,abnormal liver function,dosage of glucocorticoids,duration of glucocorticoid therapy,use of immunosuppressive agents and family history of diabetes mellitus were risk factors for SDM (all P ＜ 0.05).There were no significant differences in fasting blood glucose levels or postprandial peripheral blood glucose levels among the SDM patients receiving glucocorticoid therapy at different dosages of 0.50-0.74,0.75-0.99,1.00-1.25 mg·kg-1· d-1 (P ＞ 0.05).The peripheral blood glucose levels after breakfast,lunch and dinner were (11.50 ± 2.90),(16.02 ± 5.81) and (16.81 ± 4.52) mmol/L respectively in the patients with SDM.The levels of fasting blood glucose and glycosylated HbA 1 c were both significantly lower in the patients with SDM than in those with type 2 diabetes mellitus (t =3.74,9.92 respectively,both P ＜ 0.001).Conclusions The risk factors for SDM are age,dosage of glucocorticoids,duration of glucocorticoid therapy,fatty liver,hyperlipidemia,hypertension,abnormal liver function,use of immunosuppressive agents and family history of diabetes mellitus.The patients with SDM showed obviously elevated blood glucose levels mostly after lunch and dinner,but slightly increased levels of fasting blood glucose and glycosylated HbA 1c,which can be used to distinguish between SDM and type 2 diabetes mellitus.

LncRNA-MEG3 mediated apoptosis of retinoblastoma by regulating P53 pathway / 眼科新进展

Objective To investigate whether MEG3 involved in the development of retinoblastoma by down-regulating the expression of P53 protein.Methods The MEG3 expression of retinoblastoma tissues and corresponding non-tumor tissues were detected by quantitative real-time PCR (qRT-PCR).Retinoblastoma cell lines SO-RB50 or HXO-RB44 were transfected with pcDNA-MEG3 or siRNA-MEG3,after which cell apoptosis was tested by flow cytometry and P53 protein expression was tested by Western blot.Results MEG3 expression of retinoblastoma tissues was significantly reduced compared with corresponding non-tumor tissues(P =0.014).MEG3 level was significantly increased in pcDNA-MEG3 transfected SO-RB50 cells (P =0.002) and significantly decreased in siRNA-MEG3 transfected HXO-RB44 cells (P =0.004).Flow cytometry showed that the SO-RB50 cells apoptosis was significantly increased with the MEG3 over-expression(P ＜ 0.05),as well as the HXO-RB44 cells apoptosis was significantly decreased with the MEG3 knockdown(P ＜ 0.05),compared with the control group,respectively.Furthermore,Western blot showed that P53 protein level was significantly increased after SO-RB50 transfected with pcDNA-MEG3 (P ＜ 0.05),while significantly decreased after HXO-RB44 transfected with siRNA-MEG3 (P ＜ 0.05),compared with the control group,respectively.Conclusion MEG3 is down-regulated in retinoblastoma,affect the development of retinoblastoma,and may induce the retinoblastoma cell apoptosis by promoting the expression of P53 protein.

Effect of epigallocatechin gallate on ultraviolet B-induced transfer and degradation of melanosomes in mice / 中华皮肤科杂志

Objective To evaluate the effect of tea polyphenol epigallocatechin gallate (EGCG)on ultraviolet B (UVB)-induced skin pigmentation,transfer and degradation of melanosomes in mice,and to explore the role of autophagy in the mechanism of melanosome degradation.Methods A total of 32 ears from 16 female C57/BL6 mice were randomly and equally divided into 4 groups:acetone control group topically treated with acetone solution daily,EGCG group topically treated with 10 g/L EGCG acetone solution daily,UVB irradiation group irradiated with 500 mJ/cm2 UVB once a day and 2 hours later topically treated with acetone solution,UVB + EGCG group irradiated with 500 mJ/cm2 UVB once a day and 2 hours later topically treated with EGCG acetone solution.Ten days later,all the mice were sacrificed,and skin tissue samples were collected from the ears.Transmission electron microscopy was performed to observe ultrastructural changes of melanosomes and autophagosomes,immunohistochemical study to measure expression of protease-activated receptor 2 (PAR2) and microtubule-associated protein light chain 3 (LC3) in the epidermis,and Western blot analysis to determine the protein expression of PAR2,Rasrelated protein Rab27a and LC3 in the epidermis.Results There was a significant difference in the number of melanosomes and autophagosomes among the acetone control group,EGCG group,UVB irradiation group and UVB + EGCG group (H =12.249,13.888,respectively,both P ＜ 0.05).Compared with the acetone control group,the UVB irradiation group showed significantly increased number of melanosomes (1.85 ± 0.32 vs.1.00 ± 0.41,P ＜ 0.05)and autophagosomes (1.94 ± 0.64 vs.1.00 ± 0.46,P ＜ 0.05) in epidermal keratinocytes in mouse skin.Compared with the UVB irradiation group,the UVB + EGCG group showed significantly decreased number of melanosomes (1.30 ± 0.44,P ＜ 0.05),but significantly increased number of autophagosomes (3.03 ± 0.75,P ＜ 0.05).Immunohistochemical study showed a significant difference in the level of PAR2 in the epidermis among the 4 groups (H =18.700,P ＜ 0.05),and the expression of PAR2 was significantly lower in the UVB + EGCG group than in the UVB irradiation group (7.94 ± 4.57 vs.12.54 ± 3.07,Z =2.143,P ＜ 0.05).However,the 4 groups all showed a low level of LC3,and there was no significant difference among the 4 groups (H =5.051,P ＞ 0.05).Western blot analysis revealed significant differences in the protein expression of PAR2 and Rab27a,as well as in the LC3-Ⅱ/LC3-Ⅰ ratio,among the 4 groups (F =18.739,25.967,24.022,respectively,all P ＜ 0.05).Compared with the UVB irradiation group,the UVB + EGCG group showed significantly decreased expression of PAR2 (0.91 ± 0.54 vs.3.12 ± 0.61,P ＜ 0.05) and Rab27a (0.99 ± 0.16 vs.1.42 ± 0.07,P ＜ 0.05),but significantly increased LC3-Ⅱ/LC3-Ⅰ ratio (1.67 ± 0.08 vs.1.24 ± 0.07,P ＜ 0.05).Conclusion Topical EGCG treatment can effectively suppress UVB-induced skin pigmentation,which may be related to the inhibition of melanosome transfer and promotion of melanosome autophagy.

Observation on the clinical application of target-controlled infusion of propofol guided by Narcotrend for anesthetic induction in renal transplantation patients / 临床麻醉学杂志

Objective To observe the hemodynamic change and stress reaction of target-con-trolled infusion (TCI)of propofol guided by Narcotrend for anesthetic induction in renal transplanta-tion patients.Methods Forty patients (25 males,1 5 females,aged 21-38 years,ASA grade Ⅲ orⅣ)undergoing related living donor kidney transplantation were randomly divided into two groups:group A and group B (n =20).Group A was induced using TCI system with propofol under the moni-toring of Narcotrend.Group B was induced with propofol manually.HR,MAP,Narcotrend index (NTI),blood glucose (Glu)and plasma cortisol (Cor)were measured before induction (T0 ),before tracheal intubation (T1 ),and 1 (T2 ),3 (T3 ),and 5 (T4 )minutes afterwards.Results HR and MAP at T1 were lower than those at T0 (P < 0.05 )in two groups,they were significantly lower in group B than in group A at corresponding points(P <0.05).HR and MAP in group B increased sig-nificantly (P <0.05)and were significantly higher than those in group A (P <0.05)at T2 and T3 . There was no obvious difference in Glu and Cor between T0 and T2-T4 in group A.Glu and Cor at T2-T4 were obviously higher than those at T0 (P <0.05)in group B and those at corresponding points in group A (P <0.05).Conclusion TCI of propofol guided by Narcotrend in renal transplantation pa-tients can better control the depth of anesthesia,attenuate the stress reaction caused by tracheal intu-bation,and keep hemodynamic smooth during anesthesia induction.

Genomic diversity of the Avian leukosis virus subgroup J gp85 gene in different organs of an infected chicken

The genomic diversity of Avian leukosis virus subgroup J (ALV-J) was investigated in an experimentally infected chicken. ALV-J variants in tissues from four different organs of the same bird were re-isolated in DF-1 cells, and their gp85 gene was amplified and cloned. Ten clones from each organ were sequenced and compared with the original inoculum strain, NX0101. The minimum homology of each organ ranged from 96.7 to 97.6%, and the lowest homology between organs was only 94.9%, which was much lower than the 99.1% homology of inoculum NX0101, indicating high diversity of ALV-J, even within the same bird. The gp85 mutations from the left kidney, which contained tumors, and the right kidney, which was tumor-free, had higher non-synonymous to synonymous mutation ratios than those in the tumor-bearing liver and lungs. Additionally, the mutational sites of gp85 gene in the kidney were similar, and they differed from those in the liver and lung, implying that organ- or tissue-specific selective pressure had a greater influence on the evolution of ALV-J diversity. These results suggest that more ALV-J clones from different organs and tissues should be sequenced and compared to better understand viral evolution and molecular epidemiology in the field.

The effect of thalidomide combined with radiation therapy on growth and radiosensization on xenograft tumor of colonic carcinoma / 中国癌症杂志

Background and purpose:Thalidomide can enhance the radiation sensitivity on tumor effectively, but the mechanism of radiosensitization is still unclear. The present study aimed to investigate whether thalidomide could enhance the radiation sensitivity on colon cancer transplanted tumor of mouse, and to investigate the underlying mechanism. Methods: We established the model of colon26 colonic carcinoma, and the mice were divided into 4 groups:Control group, the thalidomide group, the radiotherapy group and thalidomide+radiotherapy group. From the day of treatment, tumors were measured every other day. Then, the xenograft tumor growth curve was depicted. Tumor volumes were measured in different treatment groups, then, the inhibitory rates of tumor growth were calcutated. Using immunohistochemical method in to detect the expression of microvessel density (MVD) in tumor tissue. Results:The mean tumor volumes at day 22 were (4.97±1.20)cm3 (control group), (2.90±0.92)cm3 (T group), (2.66±0.88)cm3 (R group), and (1.89±0.76)cm3 (T+R group). The tumor inhibition rate in the combination group (61.9%) was signiifcantly higher than the other groups (41.7%, 46.5%, P<0.05). The radiotherapy sensitization enhancement ratio of the combined treatment group was 2.27 times than in the radiotherapy group. Thalidomide combined with radiation therapy can significantly inhibit microvessel density of tumor:The decreasing MVD of T+R group, T group and R group were respectively 46.8%, 40.7%and 37.7%, and there was statistical significance between T+R group and T group (P<0.05 ), so as between T+R group and R group. It could be found more necrotic cells in tumor of group, and there was statistical signiifcance between T+R group and control group (P<0.05). Conclusion:Thalidomide can enhance the radiosensitivity mice of colonic carcinoma, and its mechanism may be related to the inhibition of tumor angiogenesis related.

Porokeratoma: a case report / 中华皮肤科杂志

A case of porokeratoma is reported.A 35-year-old man presented with asymptomatic verrucous nodules on both buttocks for more than 1 year.Skin examination at the first visit revealed two pea-sized verrucous nodules on bilateral buttocks.The lesions recurred at the primary site 3 months after surgical resection.At the second visit,there were 4 verrucous nodules,including 1 on the left buttock and 3 on the right buttock.Histopathological examination showed a well-defined lesion characterized by acanthosis and compact,thick orthokeratosis with seveal broad cornoid lamellae.A diagnosis of porokeratoma was made.The lesion was resected again followed by oral acitretin treatment.No recurrence was observed during 15 months of follow up.

Effects of baicalin on ultraviolet A-induced telomere damage in cultured human primary fibroblasts / 中华皮肤科杂志

ObjectiveTo investigate baicalin effect against ultraviolet A (UVA) induced senescence in cultured human skin fibroblasts(HSF) and influence on telomere pathway. MethodsHSF were isolated from the prepuce of neonates and cultured. Subconfluent fibroblasts were classified into blank control group (without treatment), baicalin group (treated with baicalin of 50 μg/ml), UVA group (irradiated with UVA of 10 J/cm2) and UVA + baicalin group(irradiated with UVA of 10 J/cm2 and treated with baicalin of 50 μg/ml before and after the irradiation). After additional culture of various durations, flow cytometry was performed to detect cell cycle, telomere repeat amplification protocol-enzyme linked immunosorbent assay (TRAP-ELISA) to measure telomerase activity, real-time quantitative PCR to determine telomere length, mRNA levels of p53, p16 and c-myc, Western blot to examine the protein expressions of p16 and c-myc. ResultsUVA irradiation induced cell cycle arrest in G1 phase, and the percentage of HSF at G1 phase increased from 59.94％ in the blankcontrol group to 81.04％ in the UVA group, but was decreased to 65.55％ in the UVA + baicalin group. The length of telomere in HSF in UVA group was shortened to 31.2％ of that in the blank control group, but was restored to 63.9％ in HSF treated with baicalin before and after the irradiation. Compared with the blank control group, the expression level of p53 and p16 mRNA was increased to 2.93 ± 0.21 and 2.14 ± 0.09, respectively, while that of c-myc mRNA decreased to 0.53 ± 0.03 in the UVA group; baicalin could inhibit these changes. Similarly, Western blot showed that after UVA irradiation the protein expression level of p16 increased to 5.84 ± 0.16, while that of c-myc decreased to 0.35 ± 0.04 in HSF compared with that in the blank control group; baicalin treatment before and after the irradiation induced no significant changes in the protein expres sion of c-myc, but a decline in that of p16 (4.09 ± 0.13, P ＜ 0.05). Telomerase activity was undetected in any of these groups. ConclusionsBaicalin can delay the photoaging process of HSF, which may be attributed to the regulation of expression of senescence-related genes such as p53, but not to telomerase activity.

Comparative analysis of conventional ultrasonography and contrast-enhanced ultrasound in diagnosis of renal artery stenosis / 中国基层医药

Objective To compare contrast-enhanced ultrasound and conventional ultrasound in determining artery stenosis of varying degrees of accuracy. Methods Using conventional ultrasound and contrast-enhanced ultrasound renal artery stenosis was diagnosed.And the renal artery diameter stenosis were measured the extent to which digital subtraction angiography (DSA) as the standard diagnostic accuracy of two methods to determine the rate. Results 50 patients were diagnosed as renal artery stenosis with DSA,21 patients stenosis rate 30%-49%,23 patients stenosis rate 50%-75%,6 patients stenosis rate ＞75%.diagnose accordance rate 78%(38/50)with conventional color doppler and 92% (46/50) with contrast-enchanced ultrasound.The difference was statistically significant (P＜0.05＝. ConclusionUsing ultrasound imaging of the renal artery stenosis to determine the degree had higher accuracy than the conventional ultrasonic testing method,was suitable for clinical application.

Cytochrome P450 and tumor / 中国癌症杂志

Along with the increase of the population having malignant tumors with time,the effective treatment strategy is far behind the clinical needs. There are significant individual variation in terms of treatment response and toxic profile with the same chemotherapy regimen. Therefore,the research about the occurrence and the individualized treatment of malignant tumor is one of the current hot topics. cytochrome P450(CYPs) is part of I metabolism enzymatic system,and also is the most rich in content,the most widespread in distribution and has the broadest substrate spectrum in nature. This article reviews the relationship between cytochrome P450 and the occurrence and treatment of malignant tumors.

On Strengthening Medical Ethics Education for Working Staff in Community Healthcare Service / 中国医学伦理学

Objective: To strengthen the education of ethics,honor and humiliation,and humanities for the working staff in community healthcare service,so as to better serve the community residents.Method: To strengthen the education of ethics,honor and humiliation,and humanities for the working staff in community healthcare service,and check the outcome.Conclusion: It is necessary to recognize the humanistic function of community healthcare service,strengthen practice and the ability of healthcare service to improve residents' trust and satisfaction of community healthcare service.

Prokaryotic expression,identification and purification of HCV F protein / 重庆医科大学学报

Objective:To construct a prokaryotic expression plasmid containing HCVF gene,and purify the recombinant fusion protein in E.coli system. Methods: F gene of Hepatitis C virus was amplified by PCR method from plasmid H/FL(containing full length cDNA sequence of HCV 1a subtype),cloned into pET32a(+)vector,and then transformed into E.coli JM109. After identified by restriction diges- tion and DNA sequencing,recombinant plasmid was transformed into E.coli BL21 and induced with IPTG. The fusion protein trxA- F was further confirmed by Western blot analysis and purified by affinity chromatography method. Results: Restriction digestion and PCR screening showed that HCV F gene was cloned into pET32a(+)successfully. After BL21 was transformed with recombinant vector pET32a (+)- HCVF and induced with 0.5mmol/L IPTG,a 35.4kDprotein band was found bySDS- PAGE. And this recombinant protein showed a highly specific and strong reaction with anti- His monoclonal antibody by Western blot analysis. Conclusion: The prokaryotic expression plasmid pET32a(+)- HCVF was successfully constructed,which will be helpful for further research.