OBJECTIVE：To evaluate effects of sub-chronic (90 d) exposure to cerium nitrate [Ce(NO₃)₃] on neurobehavioral functions in SD rats and to provide scientific basis for risk assessment of rare earth element cerium. METHODS：SD rats (PND21) were randomly divided into 4 groups (12 males and 12 females in each group) by body mass，which were control group (exposure to ddH₂O) and 20，100，500 mg/(kg·d) Ce(NO₃)₃ exposure groups. The rats were administrated orally with Ce(NO₃)₃ for 90 days. Neurobehavioral functions were then evaluated by experiments such as elevated plus maze， open field， rotarod and Morris water maze. Histopathological structure of hippocampus was analyzed with HE staining. RESULTS： Compared with the control group，the exposures did not cause significant changes in time on the rotarod test. From the Open field and elevated plus maze tests， there was no statistically significant differences between groups of rats in the time spent in the center of the arena，the number of entrances into the center of the arena，the percentage of entrances into the open arms and the time spent on the open arms. In the Morris water test，the difference in the escape latencies among the three cerium nitrate exposed groups and the control group was not statistically significant， nor were the differences in various metrics of the space exploration test including the times crossing platform，swimming speed and the times required for swimming in target quadrant. Histopathological analysis showed that no pathological changes were observed in the dosing groups. CONCLUSION： Sub-chronic oral exposure to cerium nitrate did not cause anxiety， deficient motor functions， and abnormal learning and memory in SD rats.

OBJECTIVE：To investigate effects of licorice and dried ginger decoction (LDGD) on tumor growth， immune organs， and liver and kidney functions in mice with breast cancers. METHODS： Female BALB/c mice were randomly divided into 5 groups：normal control group，model control group，and LDGD low-，medium-，and high-dose groups (0.1，0.3，and 0.9 g/kg，respectively). The mice in the normal control group were given distilled water by gavage. After the mice in other groups were inoculated with breast cancer 4T1 cells，the mice in the three LDGD groups were given continuous intragastric administration for 20 days from the next day， and the mice in the model control group were given corresponding volume of distilled water. The body weight，thymus index，spleen index，liver index，kidney index，serum liver function，renal function biochemical indexes， tumor volume and mass of mice in each group were measured. Pathological changes in tumor tissues from tumor-bearing mice were observed after HE staining. RESULTS：Compared with the model control group， tumor growth of 4T1 transplanted tumor mice in LDGD high-dose group was significantly inhibited， the tumor inhibition rate was (44.76 ±0.06)% (P<0.01)， the tumor inhibition rate in LDGD medium-dose group was (15.68±0.21)% (P<0.05)，and there was no obvious tumor inhibition effect in LDGD low-dose group. HE staining results of tumor histopathological sections showed that，compared with the model control group，tumor necrosis areas in LDGD high-dose group were significantly increased. In addition， LDGD at different doses had no significant effect on the body weight of mice，and indices of thymus，liver and kidney did not change significantly，but the spleen indices decreased. There were significant differences in ALP and CRE between LDGD high-dose group and normal mice，but no significant differences between LDGD high- dose group and model control group， and no significant differences in other liver， kidney and blood biochemical indices between LDGD high-dose group and normal mice. CONCLUSION：High dose LDGD had a significant inhibitory effect on tumor growth in 4T1 tumor-bearing mice，without obvious immunosuppression and hepatorenal toxicity.

OBJECTIVE： To investigate protective effects of N-acetylcysteine (NAC) on genotocicity induced by titanium dioxide nanoparticles (TiO₂-NPs) in human normal hepatocytes L- 02 and human normal lung epithelial cells HBE. METHODS：L-02 and HBE cells grown in log phase were divided into 3 groups： control group (normal culture)，TiO₂-NPs exposed group (80 μg/mL)，and TiO₂-NPs (80 μg/mL) and NAC (20 μmol/L) co-exposed group. After the cells in each group were exposed for 72 h，H₂O₂ kit was used to detect the intracellular H₂O₂ level; telomere length (TL) of L-02 and HBE cells in each group was measured by realtime quantitative PCR; chromosomal instability (CIN) and cell death indexes were evaluated by cytokinesisblock micronucleus cytome assay (CBMN-Cyt). RESULTS： Compared with the control group， TiO₂-NPs exposed for 72 h significantly increased intracellular H₂O₂ levels in L-02 and HBE cells (P<0.05)，resulting in shortened TL，increased CIN and cell death (P<0.05). When co-exposed with TiO₂-NPs and NAC for 72 h， the H₂O₂ levels and CIN among the two types cells were significantly higher than those of the control group (P< 0.05)，while TL and cell death had no significant differences compared with the control group (P>0.05). When compared with the TiO₂-NPs group，increased TL (P<0.05)，decreased CIN，and decreased cell death rates were found in TiO₂-NPs and NAC co-exposed group (both P<0.05). CONCLUSION： The antioxidant NAC strongly reduced the telomere shortening and cell death induced by TiO₂-NPs in L- 02 and HBE cells， showing the effects of protecting cells under the stress of TiO₂-NPs.

OBJECTIVE：To investigate relationships between atmospheric particulate matter pollution and non-accidental deaths among residents in Fuzhou City before and after the COVID-19 pandemic. METHODS： Daily air pollution data，meteorological data，and death registry data between 2016—2020 were collected for Fuzhou City. Time-series analysis with a quasi-Poisson regression based generalized linear model to the relationships before and after the COVID-19 pandemic after controlling for confounding factors including longterm trend，seasonality，day of week，and meteorological factors. RESULTS：After the occurrence of COVID-19，concentrations of PM_2.5 decreased significantly. Before the COVID-19，significant impacts of PM_2.5 on nonaccidental death were observed between lag1-2 days，with the greatest effects being lag day1，namely [ER= 1.69%，95%CI(0.79%，2.59%)] and [ER=0.93%，95%CI(0.49%，1.38%)]，respectively. By contrast，PM_2.5 had no significant impacts on non-accidental death after the COVID-19 pandemic. Before the COVID- 19， significant impact of PM_2.5 on respiratory-related death was observed on lag day1 [ER=3.01%，95%CI(0.35%， 5.74%)]，however，there was no significant effect after COVID-19. No noticeable changes were found on the relationships between PM_2.5 and circulatory- and cardiovascular-related deaths. CONCLUSION：Decreased PM_2.5 concentrations after COVID-19 reduced the risk of non- accidental death， particularly for respiratory-related death，indicating the necessity and effectiveness of air pollution control and prevention.

OBJECTIVE：To detect the p38MAPK gene in cell-free DNA (cfDNA) samples among Kazak esophageal cancer patients with high incidence of esophageal cancer， and the p38MAPK protein in cancer tissues， to explore their values as new molecular markers for detecting esophageal cancers. METHODS： Fasting peripheral venous blood samples from 20 Kazak esophageal cancer patients were collected before and after they had surgical treatments，and paraffin sections from the corresponding patients' postoperative tissues were collected. Blood samples from 10 healthy Kazaks were collected as controls. The cfDNA samples were extracted by the centrifugation column adsorption method and used for detection of p38MAPK via direct PCR. Expressions of p38MAPK protein in postoperative tissues were detected using immunohistochemistry and compared according to age， gender， degree of differentiation， maximum tumor diameter and T stage in clinicopathological report. RESULTS：The concentrations of cfDNA in blood of Kazakh with esophageal cancer before and after surgical resections were higher than that of healthy Kazakh (P<0.05). The detection rates of p38MAPK gene in cfDNA before operation were significantly higher than that in healthy population group (P< 0.05). However，there was no significant difference in the detection rates of p38MAPK gene in cfDNA after surgical resection between the two groups (P>0.05). Postoperative immunohistochemical detections show that the positive expression rate of p38MAPK protein in highly and moderately differentiated groups was higher than that in poorly differentiated groups (χ²=13.939， P<0.05). The positive expression rate in the group with maximum tumor diameter ≤2.5 cm was higher than that in the group with maximum tumor diameter >2.5 cm (χ²=0.014，P<0.05). There was no significant difference in the positive expression rates of p38MAPK protein between tumor tissues of patients with different age， sex and T stage(P>0.05). CONCLUSION： The concentrations of cfDNA in the blood of esophageal cancer patients were significantly higher than that of healthy people，suggesting that the concentrationscan possibly be used for early esophageal cancer screening. In addition，the detection rates of the p38MAPK gene in cfDNA of these cancer patients were significantly higher than that of healthy people，suggesting that p38MAPK gene in cfDNA can possibly be used as a potential noninvasive new molecular marker to detect the conditions of esophageal cancers.

OBJECTIVE： To investigate expressions of long-chain noncoding RNA cancer susceptibility candidate 9 (lncRNA CASC9) in colorectal cancers and its relationships with clinical prognosis. METHODS： Eighty-seven cancer and adjacent tissues from patients with colorectal cancers were collected. Real time quantitative PCR was used to detect the expression levels of lncRNA CASC9 in the tissues. Relationships between the expression levels clinicopathological indicators were analyzed. Kaplan-meier survival analysis was performed，and log-rank method was used to test the relationship between the expression levels and clinical prognosis. RESULTS：The expression levels of lncRNA CASC9 in colorectal cancer were significantly higher than that in adjacent tissues (P<0.01) and were significantly correlated with different degrees of differentiation (χ²=6.877， P=0.032)， TNM stage (χ²=7.660， P=0.022) and lymph node metastasis (χ²=9.901， P=0.002)， but not with gender，age，tumor diameter and tumor locations (All P>0.05). Kaplan Meier survival analyses show that the overall survival and progression- free survival times among patients with high expression of lncRNA CASC9 were significantly shorter than those with low expression (χ²=17.91， P<0.01； χ²=11.23， P=0.001). CONCLUSION：lncRNA CASC9 was highly expressed in colorectal cancers and was related to the degree of differentiation，TNM stage and lymph node metastasis of the cancers. It may be useful as a diagnostic marker of colorectal cancers.

OBJECTIVE： To explore correlations between EGFR mutation status and expressions of CXCR4， Ki67， MDM2 and N-cadherin in lung adenocarcinoma cells from pleural effusion. METHODS： Tumor cells from pleural effusions of lung adenocarcinomas were diagnosed using HE staining and immunocytochemistry (ICC). EGFR gene mutation status of lung adenocarcinoma patients was detected by the amplification refractory mutation system (ARMS) method，and divided into EGFR-TKI sensitive mutation group and EGFR wild- type group. Expressions of CXCR4，Ki67，MDM2 and N- cadherin were detected using the ICC technique，and the correlations between the expression levels of these proteins and EGFR mutation status were analyzed. RESULTS：In 46 patients with malignant pleural effusion，31 patients (67.4%) harbored EGFRTKI-sensitive mutations. The expression levels of CXCR4，Ki67，MDM2，and N-cadherin were correlated with EGFR mutation status (r=0.452， 0.428， 0.417， 0.524， respectively， P<0.05). CONCLUSION： Expression levels of CXCR4， Ki67， MDM2 and N-cadherin were correlated with EGFR mutation status. Furthermore， expression levels of CXCR4，Ki67，MDM2 and N-cadherin in lung adenocarcinoma cells with pleural effusions were higher in tumors with EGFR mutation than without mutation.

OBJECTIVE： To investigate expression and correlation of EGFR and HER-2 in different types of breast cancer tissues. METHODS：A retrospective analysis of 650 cases of breast cancer (600 cases of invasive ductal carcinomas， 50 cases of invasive lobular carcinoma) and 30 cases of fibroadenoma was conducted at the Surgery Treats in Handan Central Hospital from Jan. 2015 to Mar. 2020. Expression of EGFR and HER-2 were detected in 650 cases of breast cancers and 30 cases of fibroadenomas via routine detection of EGFR and HER-2 proteins by immunohistochemical method. χ² test and Spearman correlation analyses were used for statistical analysis of the data，and P<0.05 was considered statistically significant. RESULTS：The positive rate of EGFR expression in breast invasive ductal carcinoma was 48.7% which was significantly higher than that in invasive lobular carcinoma (24.0%) and fibroadenoma (16.7%) (P<0.01). The positive rate of HER- 2 expression in breast invasive ductal carcinoma was 27.8% which was significantly higher than that in invasive lobular carcinoma (8.0%) and fibroadenoma (6.7%) (P=0.002). In invasive ductal and lobular carcinomas of the breasts，the EGFR positive rate gradually increased with the increase of HER-2 score grade，and the EGFR positive rate was positively correlated with HER-2 score grade (r=1.000， P<0.05)， and the positive rate of EGFR was positively correlated with the expression of HER-2 (r=1.000，P<0.05). The positive rate of EGFR in HER-2 overexpression and TNBC was significantly higher than that in Luminal A and Luminal B， and Luminal B (HER-2 ⁺) than Luminal B (HER-2^-). CONCLUSION：The positive rates of EGFR and HER-2 were significantly increased in invasive cholangiocarcinoma of the breasts，and the positive rate of EGFR was positively correlated with the grade and positive rate of HER-2. EGFR was significantly elevated in HER-2-overexpressing and TNBC which would provide clues and evidence for targeted drug therapy for HER-2 positive breast cancers and triple negative breast carcinomas.

OBJECTIVE：To investigate effects of PM_2.5 on the migration and invasion abilities in three types of cancer cells： human lung (A549)， human liver (HepG2) and human renal (A498) cancer cells. METHODS：The A549，HepG2 and A498 cells were exposed to PM_2.5 suspension at 10 and 50 μg/mL for 24 h. The transwell assay was performed to detect cell migration and invasion abilities. RESULTS： The results from the migration assay show that the number of transmembrane A549 cells in the negative control， 10 and 50 μg/mL PM_2.5 exposure groups were (356.33±30.92)，(476.33±28.50) and (563.33±74.00)，respectively. Data for the HepG2 cells were (86.42±4.75)，(140.00±16.15) and (234.17±9.06); and those for the A498 cells were (42.33±6.43)，(80.00±6.08) and (174.67±25.01)，respectively. The results from the invasion assay show that the number of transmembrane A549 cells in the three experimental groups were (188.33±9.45)，(313.00±7.94) and (404.00±57.66)，respectively. Data for the HepG2 cells were (48.33±3.26)，(97.75±6.63) and (123.92±8.57)；and those for the A498 cells were (5.67 ±5.03)，(21.33 ±5.13) and (40.67 ±6.81)，respectively. In the migration and invasion experiments，compared with the negative control group，the numbers of penetrating cells of the three types of cells in the PM_2.5 exposure group were significantly increased (P<0.05). CONCLUSION： PM_2.5 significantly enhanced the abilities of migration and invasion in A549，HepG2 and A498 cells.

OBJECTIVE：It has been demonstrated that the single nucleotide polymorphism (SNP) in the rs3093816 site of the cyclin H (CCNH) gene was associated with an increased cancer risk. There is，however， a limitation to utilizing PCR-RFLP due to the lack of proper restriction enzyme sites in rs3093816. Therefore， an appropriate method for identifying the polymorphism was developed in this study. METHODS： A new restriction site was introduced into the CCNH amplification products using a created-restriction site PCR (CRSPCR) which allowed the enzymes CviQ I in distinguishing the PCR products. In addition，long primers were compared with relatively short primers to investigate whether the long primers are more economical and modest. RESULTS： Compared with the short primers， the larger the proportion of the long primers to the total amplified fragment， the easier to distinguish the digestion products， and the shorter the time required for electrophoresis. CONCLUSION：A mismatched long PCR primers method was successfully developed to detect the rs3093816 polymorphism in the CCNH gene.

OBJECTIVE：To investigate involvement of blood collections by time in the combined chronic toxicity and carcinogenicity test for magnesium alloy bone plate，and to provide reference for establishment of the national standard for such tests in the field of medical devices. METHODS：160 SD rats (half males and half females) were randomly assigned to the test and the control groups. The test articles were implanted into the subcutaneous tissues and sham operations were conducted to the control rats. The two groups of animals were anesthetized and blood samples were collected during the 3rd，6th，12th and 18th months for clinical pathology analyses. Hematology and clinical chemistry values were analyzed statistically. RESULTS：Compared to the control animals，there were no significant difference in hematology values and clinical chemistry values. CONCLUSION：It is feasible to cancel the blood collection by time when conducting the combined chronic toxicity and carcinogenicity test on medical devices/biomaterials. At the same time， frequencies of clinical observations could be increased so that the dying animals could be taken blood and dissected in time. This would ensure survival rates of animals and better analyses of chronic toxicity and carcinogenic effects of materials on rats.

OBJECTIVE： In vitro cytotoxicity test was used to evaluate biological safety of 23 pharmaceutical packaging materials. METHODS： The sample mass/extract volume ratio was set to be 0.2 g/mL. Extracts of pharmaceutical packaging materials were prepared with MEM medium (37 ℃，24 h)，sodium chloride injection and deionized water (121 ℃，1 h). Mouse fibroblasts L929 were cultured in a 96-well plate. 100 μL of sample extracts was added to each well and cultured for 48 hours. The absorbance was measured with a microplate reader. According to the relative proliferation of the cells (RGR) for grading，grade ≤ 1 was negative，grade 2 was suspicious positive，and grade ≥ 3 was positive. RESULTS：The 23 samples of MEM medium extracts ranged between 0-1 grade. Nine samples tested in sodium chloride injection extracts were suspicious positive above grade 2，of which 3 were positive above grade 3. The suspected positive results of 9 test samples were rechecked with deionized water extract，only 3 were suspected positive above grade 2，of which 1 was positive at grade 3. CONCLUSION：In vitro test，excluding the interference of osmotic pressure and pH value， was conducted by culture medium prepared with the maximum dissolution extract as the solvent. The approach significantly increased the detection of positive results of pharmaceutical packaging materials.

OBJECTIVE：To evaluate reproductive toxicity Capparis spinosa fruits pain gel paste in SD rats. METHODS： SD rats were randomly divided into five groups： excipient control (excipient)， positive control (cyclophosphamide) and Capparis high-dose (117.8 g/m²)，medium-dose(58.9 g/m²) and low-dose (29.5 g/m²) groups. There were 25 males and 25 females in each group. The rats were given by transdermal delivery once a day. The male rats received administration for 4 weeks before mating. Female rats received administration of 2 weeks before mating to the 6^th days after pregnancy. Positive controls received single intraperitoneal injection (0.1 g/kg cyclophosphamide) on D1 of administration in male and female rats. The rats were observed of the general condition， weight， food intake and mating during the experiment. When the female rats were confirmed to be pregnant，the male rats were executed and their epididymis，testicle and spermatozoa were examined. The female rats were executed on the 14^th day of pregnancy， and the uterus， ovaries and embryos were studied. RESULTS：There was no significant effect on weight and feed consumption of rats and pregnant rats. There were no significant effects on the mating rates，transverse diameters of testes， sperm counts of epididymis，mobility and malformation rates，epididymis and testicular organ weights. There were no significant effects on pregnancy rates， uterine fetal weights， mean corpus luteum numbers， mean implantation numbers， mean live fetus numbers， mean stillbirth numbers， mean absorbed fetus numbers， stillbirth rates and ovarian organ weights. In the high-dose group， the pre-implantation loss rates were increased (P<0.05) and the implantation rates were decreased (P<0.05). In the low- dose group， the pre-implantation loss rates were decreased (P<0.01) and the implantation rates were increased (P<0.01). These had a certain dose- effect relationship. CONCLUSION： Capparis spinosa fruits pain gel had no significant toxic effect on the fertility of male rats， while the high dose (117.8 g/m²) showed some reproductive toxicity in female rats. The medium (58.9 g/m²) and low (29.5 g/m²) doses showed no significant toxicity on female fertility.

OBJECTIVE： To investigate genetic toxicity of diphenhydramine hydrochloride and caffeine (mass ratio of diphenhydramine/caffeine is 1/2.4). METHODS：Ames test，chromosomal aberration test in CHL cells and ICR mouse bone marrow micronucleus assay were used to evaluate dphenhydramine hydrochloride and caffeine compound. In Ames test， TA97， TA98， TA100， TA102 and TA1535 were selected as indicator strains，and five test dose groups of 0.5，5，50，500 and 5 000 mg/dish were set. Chromosomal aberration test of CHL cells was divided into three dose groups，low dose (8.45 mg/mL)，medium dose (16.9 mg/mL) and high dose (33.8 mg/mL). Mouse bone marrow micronucleus assay：ICR mice were given the compounds by oral gavage. There were three dose groups：low dose (21.59 mg/kg)，medium dose (43.17 mg/kg) and high dose (86.35 mg/kg). RESULTS：Results of the Ames test show that there was no mutagenicity against Salmonella typhimurium with or without the metabolic activation system (S9). After CHL cells were exposed for 24 h and 4 h with or without S9， no chromosomal aberration effect was observed. In the mouse bone marrow micronucleus assay，the induction rates of micronucleus for the three doses were not significantly different from that of the solvent control group (P>0.05). CONCLUSION： These results indicate that diphenhydramine hydrochloride and caffeine compound had no genotoxicity based on our experimental conditions.