OBJECTIVE: To investigate effects of metformin on alleviating cadmium-induced liver injury in mice. METHODS: Male wild-type (WT) and liver-specific Ppp2r1a knockout mice (HO) were randomly divided into 4 groups： control (solvent control)， cadmium treatment (1.5 mg/kg CdCl₂)， metformin treatment (100 mg/kg metformin)， and cadmium and metformin combined treatment (1.5 mg/kg CdCl₂， 100 mg/kg metformin). These mice were consecutively administrated for 7 days by intraperitoneal injection. The semiautomatic blood biochemical analyzer was used to examine levels of ALT，AST，and TG. H&E and oil red O staining were performed to observe pathological changes of liver tissues. Real-time quantitative fluorescence PCR (qPCR) was conducted to determine mRNA levels of cadmium-related transporters. After L02 human hepatocytes were exposed to CdCl₂ and/or metformin， cell viability was detected by CCK-8 kit， and intracellular calcium ion levels were evaluated by detecting the fluorescence intensity of Fluo-4 AM under a fluorescence microscope. RESULTS: Cadmium exposure increased the blood plasma ALT and AST levels in WT mice by 86.40% and 3.19%，respectively. These mice exhibited pathological changes such as hepatocyte injury，liver lipid degeneration and deposition，and liver injury. Moreover，these effects were more profound in HO mice than that in WT mice. Compared with the cadmium treatment group alone，metformin treatment led to the 61.14% and 44.02% decrease in the plasma ALT and AST levels，respectively. Correspondingly，the degree of hepatocyte necrosis，lipid degeneration and deposition were relieved. Compared with WT mice，the mRNA levels of zip14 and dmt1， which mainly mediate cadmium transport and absorption in liver， were increased in HO mice by 16.48% and 53.61%，respectively. Upon CdCl₂ exposure，the mRNA levels of zip14 and dmt1 were increased in HO mice by 47.32% and 136.23%，respectively，compared to that in WT mice. The fluorescence intensity in L02 cells treated with CdCl₂ was increased by 2.72 times，and the corresponding cell viability was decreased by 45.99%. Compared to that in CdCl₂-treatemnt alone，1 mmol/L and 2 mmol/L metformin treatment reduced the fluorescence intensity by 40.77% and 54.17% ， respectively， and the corresponding cell viability were increased by 59.28% and 89.78%. CONCLUSION: PP2A activities were involved in CdCl₂-induced liver injury in mice by regulating the expression of cadmium transporter. Metformin could alleviate CdCl₂-induced hepatotoxicity，partly by inhibiting absorption of cadmium ions in hepatocytes.

OBJECTIVE: To investigate expression of spondin 2 (SPON2) in papillary thyroid carcinomas (PTC) and its clinical significance. METHODS: Seven fresh surgical specimens of PTC and adjacent tissues were collected. real-time quantitative PCR (qPCR) was used to detect mRNA expression of SPON2 in the tissues. From 2019-2020，68 cases of PTC paraffin specimens were collected from the Department of Pathology and expression of SPON2 protein in the specimens were detected using the immunohistochemical SP method. Prognostic relationships between SPON2 mRNA expression levels and thyroid cancers were evaluated using the Kaplan Meier-Plotter database. Relationships between SPON2 mRNA expression levels and clinical stages were analyzed using the GEPIA database. Relationships between SPON2 mRNA expression levels and thyroid cancer immune cells were analyzed using the TIMER database. RESULTS: The qPCR results showed that the mRNA expression levels of SPON2 were significantly higher in PTC tissues (1.705 ± 0.724) than in paraneoplastic tissues (0.929±0.278) (P=0.036). Immunohistochemical results showed that SPON2 protein was expressed in the cytoplasm and the extracellular matrix， and their expressions were significantly higher in PTC than in its paracancerous tissues (P<0.001). Analysis of the GEPIA database revealed that differences in SPON2 mRNA expression correlated with clinical stages of the cancers (P=0.002). The Kaplan Meier-Plotter database showed that， in 502 thyroid cancer cases， prognostic survival was worse in patients with high SPON2 mRNA expressions than with low expressions (P=0.026). TIMER database analysis showed that SPON2 mRNA expressions did not correlate significantly with thyroid cancer tumor purity (P=0.135)， but positively and significantly correlated with B cells， CD8⁺ T cells， CD4⁺ T cells， macrophages， neutrophils， dendritic cell infiltration，and the M2 macrophage markers CD68，CD163，and CD206 (P<0.001). CONCLUSION: SPON2 was highly expressed in PTC cancer tissues. Thyroid cancer patients with high SPON2 expressions had poor prognosis. SPON2 might have promoted progression of thyroid cancers through M2 macrophages and other immune cells. SPON2 may be a new marker for diagnosis of thyroid cancers， for evaluating clinical stages and prognosis of the cancers and for therapeutic target.

OBJECTIVE: To investigate the expression of HLA-E gene in human breast cancer cells and its effect on rates of proliferation，apoptosis，invasion and migration of breast cancer cells，and analyze the feasibility of siRNA (small interfering RNA) silencing of HLA-E as gene therapy. METHODS: Specific siRNA sequences of HLA-E gene were designed and synthesized，and the sequences were transfected into human breast cancer cells MDA-MB-231 with liposome Vigofect. For comparisons，blank control，negative control and liposome groups were established. Quantitative real-time PCR and Western blot were used to detect HLA-E mRNA and protein expression. CCK-8 and flow cytometry were used to detect proliferation and apoptosis rates of the cancer cells. Transwell assay was used to detect the invasion and migration. RESULTS: After transfection with HLA-E siRNA，gene and protein expressions of HLA-E in the cancer cells were reduced significantly compared with the control groups. In addition， proliferation rates of the transfected cancer cells were significantly decreased (P<0.01)，apoptosis rates were significantly increased (P<0.01) while invasion and migration were significantly decreased (P<0.01). CONCLUSION: siRNA targeting of HLA-E effectively suppressed its expression in the breast cancer cells together with inhibition of their proliferation， induction of apoptosis， and reduction of migration and invasion.

OBJECTIVE: To detect expression of the EGFR gene in lung adenocarcinomas so as to understand EGFR gene mutation in patients. METHODS: From July 1，2016 to December 1，2017，a total of 175 patients with stage III-IV lung adenocarcinomas who were admitted to our hospital were collected. Among them，all patients provided blood samples，119 provided tissue biopsy samples，175 provided plasma samples and 12 provided pleural fluid sediment cell samples. EGFR gene detection was performed on these samples using ARMS-PCR. Differences in results among the different types of samples and their relationships with patients' clinicopathological data were analyzed. RESULTS: Detection of EGFR gene expression was conducted on all blood and tissue samples. The total positive rate was 8.6% higher in the tissues than in that of blood samples and 3.1% higher than that in paraffin block tissues. EGFR mutation was found in different sex ((χ²=3.451，P=0.045)，tumor size ((χ²=8.911，P=0.002)，T stage ((χ²=17.567，P=0.000)，lymph node metastasis ((χ²=18.511，P=0.000)，metastasis station ((χ²=3.341，P=0.047) and distant metastasis ((χ²=10.874，P=0.001). EGFR mutation rates in blood and tissue samples were compared，and the difference was statistically significant (P=0.000). Tumor size [95%CI(0.151，0.725)，P=0.006]，lymph node metastases [95%CI(0.180，0.865)，P=0.020]，distant metastases [95%CI(0.097，0.612)，P=0.003] were important factors affecting progression-free survival. CONCLUSION: The combined application of samples from different tissues was useful in enhancing detection of EGFR gene expression. In addition，higher detection rate can increase the number of patients who are suitable for EGFR-tyrosine kinase inhibitors (TKIs) treatment in clinical practice. For female patients，the incidence of blood EGFR mutation was higher in those with tumor diameter ≥3 cm，stage T4，lymph node metastases with more than 3 stations，and distant metastases. Patients with tumor diameter ≥3 cm，lymph node metastasis and distant metastasis had shorter progression-free survival.

OBJECTIVE: To understand influencing factors on live-birth rates after artificial insemination with husbands-- sperm (AIH). METHODS: The study included 2391 AIH cycles from January 2011 to September 2021 in a reproductive center of the Guangdong Provincial Reproductive Science Institute. The Chi-square and the generalized estimation equation (GEE) were used to analyze factors affecting the live-birth rates. RESULTS: Inseminated women who were ≥40 years old had significantly reduced live birth rates than the younger ones ([OR=0.270，95%CI(0.0840，0.874)]). On day 3 after insemination，menstrual luteinizing hormone (LH) >8.00 IU/L，the ovulation cycle，3 eggs ovulated per cycle and prewash total motile sperm >2×10⁷ significantly increased live birth rates of insemination women [OR=1.734，95%CI(1.167，2.576)；OR=1.472，95%CI(1.037，2.090)；OR=1.567，95%CI(1.007，2.437)；and OR=2.232，95%CI(1.295，3.847)，respectively. CONCLUSION: Our data indicate that age of females，day 3 menstrual LH levels，treatment protocols，ovulation numbers per cycle and prewash total motile sperm numbers significantly affected live-birth rates after AIH.

OBJECTIVE: To identify prognostic factors among patients with stage Ⅲ gastric cancers who underwent radical resections. METHODS: From January 2017 to April 2021，116 patients with stage Ⅲ gastric cancers who underwent surgery were selected. General data and preoperative clinical indicators were collected from these patients. Correlations between serum C-reactive protein (CRP)，programmed cell death ligand 1 (PD-L1)，angiopoietin-2 (Ang-2) and clinicopathological parameters were compared. The best cut-off value of serum index before operation was determined by receiver operating characteristic (ROC) curve. Survival conditions of the patients was analyzed by Kaplan-Meier method. Cox proportional hazard regression model was established for multi-factor analysis. The value of factors in predicting prognosis of gastric cancers was evaluated using the line chart model. RESULTS: There were significant differences in preoperative CRP，PD-L1 and Ang-2 levels in age，tumor diameter，vascular invasion，nerve invasion，depth of invasion，lymph node metastasis，lymph node metastasis and differentiation. The results of survival analyses showed that the patients with preoperative serum CRPl <19.53 mg/L，PD-L1 ≥42.80 pg/mL and Ang-2 <305.16 μg/L had longer survival times and higher 1-year survival rates. The results of Cox proportional hazard regression analyses showed that preoperative CRP，PD-L1，Ang-2，tumor diameter，vascular invasion，nerve invasion，depth of invasion，lymph node metastasis and degree of differentiation were independent predictors of prognosis in patients with gastric cancers (P<0.05). The C-index consistency index of the line chart prediction model was 0.863. The calibration curve test showed that the results were highly consistent with the actual occurrence probability. CONCLUSION: In patients with gastric cancers，preoperative serum CRP levels exceeded 19.53 mg/L，preoperative serum PD-L1 levels were lower than 42.80 pg/mL，preoperative serum Ang-2 levels exceeded 305.16 μg/L，tumor diameters exceeded 5 cm，vascular invasion，nerve invasion，depths of invasion were more than T3，and those lymph node metastasis and poor differentiation had better prognosis.-Detection of preoperative serum CRP，PD-L1，Ang-2 concentration，tumor diameter，vascular and nerve invasion，depth of invasion，lymph node metastasis，differentiation and other pathological indicators can be helpful for long-term prognosis of patients.

OBJECTIVE: To investigate survival of patients with esophageal cancers，analyze prognostic factors and establish a prognostic model. METHODS: A retrospective analysis was performed on 186 patients with esophageal cancers who were admitted to the General Hospital of Veterans of Hebei Province in Xingtai，China from February 2013 to August 2017. Demographic characteristics，and clinical and pathological data of patients were collected via telephone interviews，outpatient follow-ups or inpatient reexaminations. The first date of diagnosis of a patient was designated as the starting point of the observation time，the last follow-up time or the death event of the patient was the end point，and the last follow-up time was December 30，2020. RESULTS: Among the 186 patients，162 cases (87.10%) were males，181 cases (97.31%) were Hans，144 cases (77.42%) were squamous cell carcinomas，and 135 cases (72.58%) were in clinical stages III-IV. Up to the last follow-up time，45 patients survived，and the average survival time of all patients was (33.48 ±1.72) months. The 1-，3- and 5-year survival rates were 68.89% (130/186 cases)，41.94% (78/186) and 24.19% (45/186)，respectively. Cox multivariate analyses showed that smoking history，T3-4 stage，N1 stage and M1 stage were factors for good prognosis，and surgery was a protective factor (P<0.05). Expression of the survival risk function for the patients was h(t)=h_0(t)exp (1.038×smoking history+0.419×T stage+1.062×N stage+0.772×M stage-1.100×surgical history). The larger the function index，the greater the risk. The concordance index was 0.772 with a standard error of 0.013，with high internal consistency. CONCLUSION: The 5-year survival rate of our patients was relatively low. Their prognosis was mainly affected by smoking history，T stage，N stage，M stage and surgical status. The nomogram constructed with the above indicators can be used to predict prognosis of patients with esophageal cancers.

OBJECTIVE: To investigate prognostic values of pan-Immune-inflammation index (PIV) and albumin to alkaline phosphatase ratio (AAPR) in non-small cell lung cancers with brain metastasis. METHODS: From the Second Affiliated Hospital of Zhengzhou University，the case data of 102 non-small-cell lung cancer patients with brain metastasis from December 2015 to April 2022 were retrospectively analyzed. The data included patients' basic clinicopathological characteristics，head radiotherapy status，baseline laboratory data，etc. PIV，AAPR，systemic immune inflammatory index (SII)，neutrophil/lymphocyte ratio (NLR)，derived neutrophil/lymphocyte ratio (dNLR)，platelet/lymphocyte ratio (PLR) and lymphocyte/monocyte ratio (LMR) were calculated using the blood routine and liver function test results. The best cut-off values of PIV，AAPR，etc for predicting the prognosis of the cancer cases were calculated using the X-tile software. χ² test or Fisher--s-exact-test-was used to evaluate correlations between PIV，AAPR and various clinicopathological indicators. Univariate and multivariate Cox proportional hazard regression models were used to analyze possible correlations between these variables and clinical outcomes，and to explore factors affecting the survival of the patients. Survival curves were drawn using the R software. RESULTS: The cutoff values for PIV，AAPR，SII，NLR，dNLR，PLR，and LMR were 221.9，0.6，1 039.4，5.2，4.0，223.7，and 4.5，respectively. The univariate Cox regression model showed that the survival curves of PIV，AAPR，SII，NLR，dNLR，LMR in different levels were statistically significant (P<0.05). The multivariate Cox regression model showed that PIV and AAPR had a certain impact on the prognosis of patients (P<0.05). CONCLUSION: Our results show that high PIV (≥221.9) and low AAPR (<0.6) were related to poor clinical outcomes. Thus，PIV and AAPR can be used as biomarkers to predict prognosis of non-small cell lung cancer patients with brain metastasis.

OBJECTIVE: To detect changes of inflammatory factors in intraepithelial colorectal adenomas，and to investigate correlations between the factors and gradings of the adenomas. METHODS: Clinical data from 200 cases of patients with adenomas were retrospectively analyzed. The adenomas were divided into different groups according to their neoplasia grades. Age，sex，adenoma location，number of adenomas，maximum adenoma diameter，counts of neutrophils，platelets，monocytes and lymphocytes，mean platelet volumes，ratios of platelets to lymphocytes (PLR)，neutrophils to lymphocytes (NLR) and monocytes to lymphocytes (MLR)，systemic Immune-inflammation indices (SII)， and carcinoembryonic antigens (CEA) were analyzed. RESULTS: The AUC of PLR，NLR，SII，MLR were 0.614，0.653，0.656 and 0.593，respectively. The optimal cut-off values of PLR，NLR，SII，MLR were 165.01，1.87，465.71 and 0.26，respectively. Univariate analyses showed that PLR，NLR，SII，MLR，CEA，neutrophil count、lymphocyte count，adenoma location，maximum adenoma diameter were significantly related to the neoplasia grades of the adenomas. According to the binary Logistic regression analyses，PLR，SII and maximum adenoma diameters were the risk factors for high-grade intraepithelial neoplasia. CONCLUSION: The higher the platelet-to-lymphocyte ratios and systemic inflammatory response indices in the peripheral blood of patients with colorectal adenomas，the greater the likelihood of high-grade intraepithelial neoplasia. Their positive correlations may be useful for enhancing therapy for colorectal adenoma.

OBJECTIVE: To understand impacts of PM_2.5 and PM₁₀ concentrations in operating rooms from smog which was produced from electrosurgical knife usage. METHODS: 50 cases with thoracolumbar fractures who needed electrosurgical knife usage and 50 cases who did not require electrosurgical knife for vertebroplasty were selected as the observation and the control groups，respectively. Air samples in the clean laminar flow operating rooms were collected to determine PM_2.5 and PM₁₀ concentrations. RESULTS: Concentrations of PM_2.5 and PM₁₀ in the air of the observation group were significantly higher than those in the control group (P<0.05). CONCLUSION: The smog from electrosurgical knife usage was a cause of pollution in the operating rooms which posed risks of health hazards for both medical professionals and patients.

OBJECTIVE: To use the Ames and the HPRT tests，six oxidative hair dyes were tested，and application of the two methods in detecting mutagenicity of oxidative hair dyes was discussed. METHODS: Four dose groups，ranging from 312 to 10 000 μg/dish were set up for the Ames test. mutagenicity was determined by calculating the numbers of reverted mutant colonies. In the HPRT test，V79 cells were cultured in vitro，and 4 dose groups were set up for analysis，with the dose range from 4.88 to 5 000 μg/mL，to determine mutation frequencies of the HPRT sites. RESULTS: With or without an in vitro metabolic activation system in the Ames assay，the numbers of retroactive colonies in the treated groups were more than twice that of the solvent control group，and there was a dose-effect relationship. However，there was no statistically significant difference in cell mutation frequencies between the treated and the control group with or without the metabolic activation system. CONCLUSION: Due to cytotoxicity of the oxidative hair dyes to mammalian cells，the HPRT test was not capable of detecting mutagenicity of the hair dyes. Thus，the Ames test which is simple，rapid and economical，and can be used as the first choice for mutagenicity test.