OBJECTIVE: To observe the photobleaching characters of sinoporphyrin sodium (DVDMS) in different solutions and skins of animal. METHODS: Absorption peaks of DVDMS (10, 20, 40 μg/mL) in normal saline (NS), RPMI-1640 medium (RPMI-1640), RPMI-1640 medium containing 10% fetal calf serum (RPMI-FBS) and suspension of HaCaT cells were measured using micro-spectrophotometer. Using DVDMS solutions with concentrations of 1, 2.5, 5, 10, 20 and 40 μg/mL in the 4 solvents above, photobleaching of DVDMS after laser radiation for 3, 6, 10, 20, 40, 50, 60, 120, 180 min was observed using microplate reader. In vivo image acquisition system was used to study the distribution of DVDMS in animal skins, and the bleaching after irradiation in psoriasis-like animal skins were also observed. RESULTS: DVDMS with concentrations 10, 20 and 40 μg/mL showed obvious characteristic absorption peaks in the 4 solutions. Photobleaching occurred in the 4 solutions under the irradiation conditions used in previous pharmacodynamics studies (laser radiation was 3 minutes). Two-phase processes photobleaching (fast first phase and second slow phase) of DVDMS were observed in 180 min. The distributions of DVDMS in skins of normal and psoriasis-like animal were uniform, obvious photobleaching occurred in skins of psoriasis-like animal after laser irradiation with dose of 9.6 J/cm². CONCLUSION: In vitro photobleaching of DVDMS was influenced by light dose, initial concentrations and solvents. Secondary-dynamic model was suitable to describe the photobleaching progress. Photobleaching occurred in skins of psoriasis-like animal after laser irradiation.

OBJECTIVE: To understand autophagy induction by tert-butyl hydroperoxide (t-BHP) in human hepatic cell line L02 and to explore the involvement of mitochondria. METHODS: L02 cells were treated with different concentrations of t-BHP (100, 200, 400, 800, 1000 μmol/L) with or without specific inhibitors (mitochondria targeted antioxidant MitoQ or p38/MAPK inhibitor SB203580). Autophagy markers LC3-Ⅱ and p62 as well as the p38/MAPK pathway proteins were detected by immunofluorescence and/or Western blot. Mitochondrial reactive oxygen species (ROS) were measured by flow cytometry with Mito-SOX Red kit. RESULTS: The high concentrations of t-BHP (≥800 μmol/L) induced the accumulation of LC3 fluorescence puncta, with LC3-Ⅱ protein level increased and p62 protein level decreased significantly. These changes accompanied the elevated mitochondrial ROS and the activated p38/MAPK pathway. Pretreatment with mitochondria targeted antioxidant MitoQ or specific p38/MAPK inhibitor SB203580 attenuated these changes which were induced by t-BHP (1000 μmol/L). CONCLUSIONS: We established the autophagy model induced by t-BHP in human hepatic cell line L02. Mitochondrial ROS and p38/MAPK pathway played critical roles in this process.

OBJECTIVE: To understand the relationship between aberrant expression of E-cadherin in nasopharyngeal carcinoma (NPC) and clinicopathological features of NPC patients for clinical applications. METHODS: International publications about E-cadherin and NPC were selected from the Cochrane Library, Pubmed, EMbase database, and Chinese publications were from CBM, CNKI. All analyses were performed by software STATA 11.0. Abnormal expression, degree of differentiation, T stage, lymph node metastasis, and clinical stage were analyzed using pooled odds ratio (OR) with 95% confidence interval (CI). RESULTS: A total of 16 studies including 1040 patients were enrolled in the Meta-analysis. It was shown that expression of E-cadherin was significantly increased in NPC compared to that in normal tissues (OR=0.096, 95% CI:0.036-0.252, P=0.000). In addition, the aberrant expression of E-cadherin was significantly associated with the degree of differentiation, lymph node metastasis and clinical stages (OR=0.232, 95% CI:0.061-0.884, P=0.032;OR=0.324, 95% CI:0.198-0.53, P=0.000;OR=0.441, 95% CI:0.232- 0.835, P=0.012). CONCLUSION: NPC often showed aberrant expression of E-cadherin. In addition, patients with the aberrant expression were prone to having tumors at lower differentiation, with lymph node metastasis and at late clinical stage. E-cadherin may be an indicator for differentiation, lymph node metastasis and clinical stage.

OBJECTIVE: To study the effect of vinorelbine tartrate lipid microsphere (NVB-lip) injection on growth of human breast cancer cells in nude mice. METHODS: 35 female BALB/c-nu mice were inoculated with human breast cancer BCAP-37 cells. These mice were divided randomly into 5 groups for further treatments:NVB-lip high dose group (NVB-lip 10 mg/kg), medium dose group (NVB-lip 5 mg/kg), low dose group (NVB-lip 2.5 mg/kg), positive drug control group (NVB-free 5 mg/kg) and negative control group (only lipid microsphere). There were 7 mice each group. Injection with NVB-lip was done after 6-8 days when the tumors shaped. NVB-lip was injected intravenously every 3 or 4 days each week for 6 times in total. Weights and tumor volumes(V_T) of mice were observed and measured on No. 4, 7, 11, 14, 18 and 23 days from the first injection. Relative tumor volumn(V_RT), relative tumor proliferation rate and tumor inhibition rate were calculated. RESULTS: The weights of nude mice for different groups increased slowly but the differences were not statistically significant. Compared with the negative group, the (V_RT) and relative tumor proliferation rates of NVB-lip groups were reduced but tumor inhibition rate was increased. The differences were statistically significant (P<0.01). CONCLUSION: The inhibition of NVB-lip high dose group(2.5-10 mg/kg) was more obviously than that of the other groups. NVB-lip may have clinical applications.

OBJECTIVE: To investigate the expression level of miR-338 cluster in esophageal carcinoma and identify the relationship between miR-338 cluster and esophageal carcinoma. METHODS: A total of 86 cases of patients with confirmed esophageal carcinoma were selected, real time RT-PCR were performed to detect the expression of hsa-miR-338-3p, hsa-miR-338-5p and hsa-miR-657. We used paired t-test to analyze the expression of miRNA in tumor and adjacent non-tumor tissues, Pearson correlation analysis to examine the correlation of the expression of each miRNA and logistic regression analysis to analyze the effect of abnormal expression of miRNAs on the risk of esophageal cancer. RESULTS: hsa-miR-338-3p and hsa-miR-338-5p were downregulated in tumor tissues compared with adjacent non-tumor tissues (P<0.05), and no statical difference was found of the expression of hsa-miR-657 between the two tissues (P>0.05). The expression of hsa-miR-338-3p and hsa-miR-338-5p were significantly correlated (r=0.754, P<0.05), and low expression of hsa-miR-338-5p was the risk factor for esophageal carcinoma (OR=1.264, P<0.05). CONCLUSION: miR-338 cluster may play an important role in the formation process of esophageal carcinoma. In addition hsa-miR-338-5p may represent the miR-338 cluster to be a useful biomarker for diagnosis.

OBJECTIVE: To investigate the effect of stem cell-related factor, Notch3 on human triple negative breast cancer cell, MDA-MB-231 and its resistance to cisplatin, and explore if overexpressed Notch3 can enhance the sensitivity of resistance cells to cisplatin. METHODS: Lipofectamine 2000 was applied to transfect Notch3 plasmid into MDA-MB-231. These cells were divided into MDA-MB-231-PCLE group and MDA-MB-231-N3ICD group, according to the use of different plasmids. RT-PCR and Western blot methods were used to analyze the expression of Notch3 in different groups. The effects of cisplatin on cell proliferation were assessed by CCK8 assay and evaluated by GraphPad Prism 5. Lipofectamine 2000 was also applied to transfect the siNotch3 or scramble to cells and they were divided into T47D-NC and T47D-siNotch3 groups. The protein levels of apoptosis-related factor were detected using RT-PCR methods and Western blot. RESULTS: Then efficacy of cisplatin to triple negative breast cancer cells was enhanced by overexpression of Notch3. Apoptosis-related factor, caspase-3 was up-regulated by Notch3 overexpression. However, siNotch3 induced the downregulation of caspase-3 in T47D, in which the expression of Notch3 was high. CONCLUSION: Overexpression of Notch3 enhanced the sensitivity of MDA-MB-231 to cisplatin which suggests that this may improve the efficiency of chemotherapy in patients with breast cancer.

OBJECTIVE: To investigate theinduction of AGR2 expression by dinitrosopiperazine (DNP) in nasopharyngeal cancer cell (NPC) 6-10B and to explore involvement of DNP in NPC metastasis. METHODS: Non-cytotoxic concentrations of DNP on 6-10B cells were determined using MTT. Expression of AGR2 protein and mRNA in 6-10B cells which were treated with DNP were detected by indirect immunofluorescence, Western blotting and real-time RT-PCR, respectively. In addition, silencing the expression of AGR2 by siRNA-AGR2 was investigated. The capabilities of invasion and migration of 6-10B cells were investigated using Transwell assay. RESULTS: The range of non-cytotoxic concentrations (NCC) of DNP was 0-10.0 μmol/L. The results of indirect immunofluorescence showed AGR2 mainly expressed in cytoplasm after DNP treatment. These and 5-8F cells had stronger fluorescence intensity than untreated 6-10B cells. The DNP-induced AGR2 expression showed dose- and time-dependent effects. Real-time RT PCR analyses showed that AGR2 mRNA expression rate was 1.01±0.08 in the untreated 6-10B cells and 3.96±0.15 in the cells treated with 8.0 μmol/L DNP. The difference was significant. In addition, the relative fold gene expression of AGR2 in 5-8F cells was higher than that in non-treated 6-10B cells (P<0.05). The transwell migration and invasion data showed that siRNA-AGR2 transfection blocked AGR2 expression in 6-10B cells. In addition, both DNP-induced invasion and motility were dramatically decreased when AGR2 expression was blocked (P<0.05). However, DNP could effectively induce cell invasion (P<0.05) and motility (P<0.05) when AGR2 was not blocked. CONCLUSION: DNP could up-regulate AGR2 expression in 6-10B cells through transcriptional regulation mechanism and then mediate the metastasis of nasopharyngeal cancer cells.

OBJECTIVE: To investigate whether any of the three polymorphisms (rs2274083, rs2274084, rs72474224) of GJB2 are associated with susceptibility to occupational noise-induced hearing loss in Chinese Han population. METHODS: A 1:1 case-control study was conducted using 177 cases with average hearing threshold of no less than 40 dB in high frequency and 177 controls with average hearing threshold of less than 25 dB in high frequency. Controls were matched with cases according to gender, age, duration of noise exposure and working positions. The genotypes were sequenced after polymerase chain reaction. RESULTS: There was statistically significant differences in the distribution of genotypes and alleles frequencies of rs2274084 between cases and controls. The rs2274084 C and T allele frequencies in cases and controls were 73.73%, 26.27% and 63.84%, 36.16%. The CC genotypes, TC genotypes, TT genotypes in cases and controls were 55.37%, 36.72%, 7.91% and 40.11%, 47.46%, 12.43% respectively. There was no statistically significant difference in the distribution of genotypes of rs2274083 and rs7274224 between cases and controls. CONCLUSION: The data suggest that GJB2 rs2274084 might be the susceptibility gene locus of noise-induced hearing loss in Chinese workers. In addition, workers with the C genotype in rs2274084 might be susceptible to noise-induced hearing loss.

OBJECTIVE: To identify a better maintenance medium for primary hepatocyte culture in vitro, we compare the cellular morphology and function of mouse primary hepatocytes under three different culture conditions. METHODS: Primary hepatocytes were isolated by using a modified two-step collagenase perfusion procedure as described by Seglen. The purity of hepatocytes were determined by PAS staining. After plating for 4 h, the medium was replaced with three different maintenance medium (base medium, base medium supplemented with DMSO, base medium supplemented with DMSO and EGF). The cellular morphology was observed under an inverted microscope. The cell viability was measured using the MTT assay, the concentrations of lactate dehydrogenase (LDH) was detected by automatic biochemical analyzer and the albumin level was examined using ELISA. RESULTS: Mouse primary hepatocytes were successfully isolated with purity more than 95%. When primary hepatocytes were cultured for 96 h, only the DMSO group had better cell morphology. Viability of cells in the base medium group was reduced by 66.87% and 67.16% compared with the DMSO and the DMSO+EGF groups (P<0.05), respectively. The LDH level of the base medium group was higher than the DMSO and the DMSO+EGF groups (P<0.05). In addition, cells in the DMSO group had high level of albumin secretion at 96h which was 185% and 24.2% higher than the base medium and the DMSO+EGF group (P<0.05), respectively. CONCLUSION: DMSO addition to maintenance medium supported primary hepatocytes to maintain its morphology and function. Our study provides an applicable method for optimization of primary hepatocytes in culture.

OBJECTIVE: In humans, aneuploidies occur in at least 5% of all clinically recognized pregnancies, among which about 90% can be caused by maternal meiotic error. In this study, a stable and simple method for fluorescence in situ hybridization (FISH) with human oocyte chromosomes was developed to explore causes of maternal meiotic error, mechanisms of aneuploidy and genetic effects of environmental factors on human oocytes. METHODS: The abandoned human oocytes were collected from IVF patients who signed consent forms. The oocytes were used to prepare chromosomal karyotype through hypotonic and gradual fixative treatments. Human oocyte metaphase FISH was performed using the CEP 16 Probe. RESULTS: From our observations, the morphology and dispersal of human oocyte chromosomes were excellent, and the fluorescence signals were bright and clear with clean background. CONCLUSION: We established successfully the method for FISH with human oocyte chromosomes.

OBJECTIVE: To investigate whether the polymorphisms of Arg194Trp on X-ray repair cross-complementing group 1(XRCC1) would influence susceptibility to colorectal cancer among Chinese. METHODS: We collected publications which discussed association between polymorphisms of XRCC1 Arg194Trp and susceptibility to colorectal cancer in Chinese people by searching the databases (wanfangdata, CBM, CNKI, VIP, PubMed, EBASE). The pooled ORs with their 95%CI were calculated by using Review Manager 5.1 software. We also conducted the heterogeneity and sensitivity, publication bias and subgroup analyses. RESULTS: After filtering, 10 studies were included on the selection criteria. The result of this Meta-analysis demonstrates:homozygous model, OR=1.40, 95%CI (1.13-1.73), P=0.002;dominant model, OR=1.22, 95%CI(1.01-1.47), P=0.04;recessive model, OR=1.30, 95%CI (1.06-1.60), P=0.01;heterozygous model, OR=1.19, 95%CI(1.00-1.42), P=0.05. In the subgroup analysis, the value of OR between smokers in case group and smokers in control group is 1.04, 95%CI(0.84-1.29), P=0.72;the value of OR between drinkers in case group and drinkers in control group is 1.08, 95%CI(0.81-1.44), P=0.60. CONCLUSION: The polymorphisms of Arg194Trp on XRCC1 contributed to susceptibility of colorectal cancer. Those who carried the 194Trp genotype had increased risk of colorectal cancer in the dominant, homozygous and recessive models. Smoking and drinking had no significant correlation with the incidence of colorectal cancer.

OBJECTIVE: The micronucleus test with Vicia faba root tip cells was used to study genotoxicity of pollutants from rivers around nuclear facilities. METHODS: 7 samples collected from river of Furong bridge, Fujiang, Anchang, etc. were used to treat V. faba root tip cells directly. Micronuclei frequencies (MNF) and proliferation index (PI) were determined and statistics analyses were performed to evaluate the difference in MNF among different samples. RESULTS: There was significant differences in MNF for every site. PI values of 5 sites among 7 sampled sites were over 2, and it is highly significant compared with the contrast one (P<0.01). CONCLUSION: The results show that the sampling points of radiation pollution levels were lower than the national standard limit value. However, water pollution of the rivers around the nuclear facilities was mainly caused by cross-strait industrial, life, and gabage pollution. Radioactive effluent generated by nuclear facilities operation affected the public and the environment lightly.

OBJECTIVE: To explore the gene mutation types and proportions of non-deletion α-thalassemia, and to provide clinical basis for genetic counselling. METHODS: The gene mutation types and frequencies of 259 suspected non-deletion α-thalassemia patients were analyzed by Gap-PCR and PCR-RDB. 30 pregnant women whose husband had the same type of thalassemia received thalassemia prenatal diagnosis. We completed follow-up works. RESULTS: Among the 259 cases, 170 cases of αα^QS/αα, 43 cases of αα^WS/αα, 34 cases of αα^CS/αα, 6 cases of——^SEA/αα^QS, 4 cases of——^SEA/αα^CS, one case of -α^3.7/αα^QS, and one case of -α^4.2/αα^QS were found. Compared with normal control group, there were statistical significances in MCV, MCH in different types of non-deletion α-thalassemia (P<0.05). 5 fetuses with non-deletion HbH disease were confirmed by prenatal diagnosis. The results of postpartum follow-up were consistent with prenatal diagnoses. CONCLUSION: Guangzhou is a high incidence area of non-deletion α-thalassemia. The most common genotype is αα^QS/αα. To reduce the birth of children with non-deletion HbH disease, non-deletion thalassemia genotype carriers should be tested.

OBJECTIVE: To investigate toxin contaminations of shellfish products in Shenzhen city from 2011 to 2014. METHODS: We collected 386 samples of 23 types of shellfish from six districts of Shenzhen city. We tested and evaluated the levels of paralytic shellfish poison (PSP), diarrheic shellfish poison (DSP), neurotoxin shellfish poison (NSP) and amnesic shellfish poison(ASP) in the samples by enzyme-linked immunosorbent assay (ELISA). RESULTS: There were 78 samples exceeded limits in 386 samples with a over-limit ratio of 20.21%, among which PSP was 3.11% (12/386) and DSP was 18.13%(70/386). But NSP and ASP were both normal. There were 16 kinds of shellfish with DSP and 3 kinds of shellfish with PSP over limits. DSP contamination was serious in Ruditapes philippinarum, Scapharca subcrenata and Ostrea gigas thunberg, while PSP excess was mostly in Circle Shellfish. CONCLUSION: In recent years DSP and PSP were contaminants in shellfish products in Shenzhen city. We need to strengthen health supervision and guarantee consumer food safety.