[Overexpression of tuftelin and KLF-5 and its clinicopathological features in hepatitis B virus-related hepatocellular carcinoma].

Objective: To analyze and evaluate the expressions and clinical value of tuftelin (TUFT1) and Krüppel-like factor 5 (KLF5) in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) tissues. Method: KLF5 mRNA and TUFT1 mRNA transcriptional status in cancer and non-cancer groups were compared according to the Cancer Genome Atlas (TCGA) database. The differences and prognostic value between the groups were analyzed. Postoperative liver cancer and its paired pericancerous tissues, with the approval of the ethics committee, were collected to build tissue chips. The expression of KLF5 and TUFT1 and their intracellular localization were verified by immunohistochemistry. Tissue expression and clinicopathological characteristics were analyzed by immunoblotting. SPSS software was used to analyze the relationship between SPSS and patient prognosis. Results: The transcription level of TUFT1 or KLF5 mRNA was significantly higher in the HCC group than the non-cancer group (P < 0.001), according to TCGA data. Immunohistochemistry and Western blotting examination confirmed the overexpression of TUFT1 and KLF5 in human HCC tissues, which were mainly localized in the cytoplasm and cell membrane. The positivity rates of TUFT1 and KLF5 were 87.1% ( χ(2) = 18.563, P < 0.001) and 95.2% ( χ(2) = 96.435, P < 0.001) in HCC tissues, and both were significantly higher than those in the adjacent group. The expression intensity was higher in stage III-IV than stage I-II of the International Union Against Cancer standard (P < 0.01). The clinicopathological features showed that the abnormalities of the two were significantly related to HBV infection, tumor size, extrahepatic metastasis, TNM stage, and ascites. Univariate analysis was related to tumor size, HBV infection, and survival. Multivariate analysis was an independent prognostic factor for patients with HCC. Conclusion: TUFT1 and KLF5 may both be novel markers possessing clinical value in the diagnosis and prognosis of HBV-related HCC.

[Research progress on the relationship between liver cirrhosis and thyroid hormones].

There exists a complex relationship between liver and thyroid hormones. Liver plays an important role in the activation, inactivation, transportation, and metabolism of thyroid hormones. At the same time, thyroid hormones also affect hepatocytes activity and liver metabolism, such as lipid and bilirubin metabolism. Importantly, thyroid hormone levels often change abnormally in patients with liver cirrhosis. Therefore, studying the change of thyroid hormone levels in patients with liver cirrhosis has a certain clinical value for assessing the severity, prognosis, diagnosis and treatment. This paper reviews the research progress on the relationship between liver cirrhosis and thyroid hormone.

[The effect of Shendan Sanjie capsule on angiogenesis in mice with colitis associated cancer and mechanism].

Objective: To investigate the effects and the mechanism of Shendansanjie capsules on angiogenesis of colitis associated cancer(CAC) mice. Methods: Azoxymethane and dextran sulfact sodium were used to construct a mice model with CAC. Ten mice were divided into the normal group, model group, Shendan Sanjie capsule group, MK-2206 group, and Shendan Sanjie capsule + IGF-1 group, respectively. Immunohistochemistry was used to detect the microvessel density (MVD) in the colon tissue of each group of mice. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the mRNA levels of basic fibroblast growth factor (bFGF) and angiopoietin 2 (Ang2) in colon tissue. Western blot was used to detect the expressions of Akt, p-Akt, vascular endothelial growth factor A (VEGFA), hypoxia-inducible factor-1α (HIF-1α). Results: The number of MVD in the colon tissue of mice in the model group, Shendan Sanjie capsule group, MK-2206 group, Shendan Sanjie capsule + IGF-1 group were 63.3±3.3, 36.6±2.3, 36.6±2.2, 50.3±2.5, significantly higher than 2.0±0.1 in the normal group (P<0.05). The number of MVD in Shendan Sanjie capsule group, MK-2206 group and Shendan Sanjie capsule+ IGF-1 group are lower than that in model group (P<0.05), while Shendan Sanjie capsule+ IGF-1 group is higher than Shendan Sanjie Capsule group (P<0.05). The relative expressions of bFGF mRNA in the colon cancer tissue of mice in the model group, Shendan Sanjie capsule group, MK-2206 group and Shendan Sanjie capsule+ IGF-1 group were 4.55±0.31, 2.46±0.37, 2.49±0.33, 3.34±0.21, respectively, and the relative mRNA expressions of Ang2 were 5.78±0.19, 2.21±0.14, 2.26±0.17 and 3.67±0.32, respectively, which were significantly higher than 1.01±0.05 and 0.99±0.07 in the normal group (P<0.05). The mRNA levels of bFGF and Ang2 in Shendan Sanjie capsule group, MK-2206 group and Shendan Sanjie capsule+ IGF-1 group were lower than those in the model group (P<0.05), while Shendan Sanjie capsule+ IGF-1 group is higher than Shendan Sanjie capsule group (P<0.05). The relative expression levels of p-Akt/Akt, VEGFA and HIF-1α in colon cancer tissues of the model group were 4.75±0.18, 4.64±0.22 and 4.84±0.12, respectively, which were significantly higher than 1.01±0.07, 0.95± 0.08 and 0.98±0.05 in the normal group (P<0.05). The relative expressions of p-Akt/Akt, VEGFA and HIF-1α in colon cancer tissues in the Shendan Sanjie capsule group were 2.24±0.22, 3.15±0.26 and 2.07±0.18, respectively, which were significantly lower than those in the model group (P<0.05). However, compared with the MK-2206 group, the difference was not statistically significant (P>0.05). The relative expression levels p-Akt/Akt, VEGFA and HIF-1α in colon cancer tissue of the Shendan Sanjie capsule+ IGF-1 group were 3.37±0.15, 4.02±0.11, 3.52±0.24, respectively, which were significantly higher than those in the Shendan Sanjie capsule group (P<0.05). Conclusion: Shendan Sanjie capsules may inhibit Akt/HIF-1α/VEGFA signaling pathway, and then reduce the expression of microvascular growth factors bFGF and Ang2, thereby inhibit the tumor angiogenesis of CAC.

[Relationship of C1QA level and therapeutic effect and prognosis of DLBCL patients treated with R-CHOP].

Objective: To investigate the relationship between plasma levels of complements before treatment and the clinicopathological feathers and prognoses of diffuse large B-cell lymphoma (DLBCL) patients treated with Rituximab (R)-CHOP or R-CHOP-like therapy. Methods: The clinicopathological data of 105 DLBCL patients treated in cancer Hospital of Chinese Academy of Medical Sciences from 2010 to 2016 were collected. The plasma samples from 105 DLBCL patients treated with R-CHOP or R-CHOP-like therapy and 80 healthy controls were used to detect 34 complement levels before treatment by utilizing antibody microarray. The relationship between plasma levels of complements and the clinicopathological feathers and prognosis of DLBCL patients were analyzed. Results: The signal values of C1QA and CR1L in patients with international prognostic index (IPI) scores of 3-5 were 1 261.43±138.9 and 2 214.69±98.58, respectively, higher than 950.79±80.19 and 984.67±121.79 in patients with IPI scores of 0~2 (both P<0.05). The levels of C1QA and CR1L in the non-complete response (CR) group were 1 165.43±98.56 and 2 263.13±145.63, respectively, higher than 914.70±100.77 and 1 821.34±84.68 in the CR group (both P<0.05). Cox regression analysis showed that elevated C1QA signal value was associated with poor progression-free survival (PFS) and poor overall survival (OS) (PFS: HR=2.063, 95%CI: 1.220-3.489, P=0.007; OS: HR=2.23, 95%CI: 1.036~4.798, P=0.040). After IPI correction by Cox multivariate model, the elevated C1QA signal value was still correlated with poor PFS (HR=1.765, 95%CI 1.034~3.013, P=0.037). Conclusions: The baseline plasma levels of C1QA and CR1L are correlated with IPI scores and therapeutic effects of DLBCL patients treated with R-CHOP. The baseline plasma level of C1QA has a certain predictive value for the prognostic evaluation of DLBCL.

[Reliability and validity test of Chinese version of the Karitane parenting confidence scale].

Objective: To translate and revise the Karitane Parenting Confidence Scale (KPCS),which can be used to evaluate the parenting confidence of 0-12 months infant caregivers in China, and evaluate the reliability and validity test of Chinese version of KPCS. Methods: Form a Chinese version of Karitane Parenting Confidence Scale through translation, back translation and expert review. Mothers of 3-month-old infants were recruited from two Maternal and Child Health Hospitals in Beijing and Ma'anshan in April 2019. A total of 165 mothers responded the survey invitations. They were surveyed with self-administered questionnaires, the Chinese version of KPCS, the Parenting Sense of Competence Scale (PSOC) and Self-efficacy in Infant Care Scale (SICS). Item analysis was conducted to select items by using critical value and correlation coefficient. The construct validity was assessed by exploratory factor analysis, confirmatory factor analysis. The criterion validity was assessed by being compared with PSOC and SICS. The reliability analysis was assessed by Cronbach's α the split-half reliability coefficient and rest-retest reliability coefficient. Results: The scores of 15 items were all correlated with the total score of the Chinese version of KPCS with r ranging from -0.283 to 0.643 (P<0.001). The difference of critical values of all items of KPCS among the low and high score groups were statistically significant (P<0.001). Three factors labeled parenting, support, and sense of competence, were obtained by exploratory factor analysis which accounting for 49.52% of the total variance and the factor loading values of all items are more than 0.4. The confirmatory factor analysis confirmed the hypothesized three-factor structure. The total score of KPCS was significantly correlated with the total score of PSOC and SICS(r=0.381, 0.345, P<0.001). The Cronbach's α of the Chinese version of KPCS was 0.769, and each dimension of Cronbach's α were 0.332-0.800, the test-retest reliability coefficient was 0.817, and the split-half reliability coefficient was 0.789. Conclusion: The Chinese version of the Karitane parenting confidence scale has a good reliability and validity among the 0-12 month-old infants' mothers, which can be used to evaluate the parenting confidence of infant caregivers.

[Studies of the norm of Karitane Parenting Confidence Scale(KPCS)among parents of infants in urban areas of China].

Objective: To establish the norm of the Chinese version of Karitane Parenting Confidence Scale (KPCS) in urban areas of China. Methods: From August to December 2017, the parents of 2 216 children (<36 months old) were selected from 15 cities (Beijing, Lianyungang, Hangzhou, Chengdu, Xi'an, Guangzhou, Changsha, Jinan, Guiyang, Ningbo, Dalian, Qinhuangdao, Maanshan, Chongqing and Wuhan) in 14 provinces by stratified random sampling. The general demographic characteristics and parents' parenting confidence were collected by a self-made questionnaire and KPCS Chinese version. The percentile norm was established. P3, P10 and P25 were used as the criteria to define the degree of lack of parenting confidence. Results: The age of mothers was (30.67±4.29). The age of the father was (32.50±4.99) years old. There were 726 (32.76%), 759 (34.25%) and 731 (32.99%) infants in 6-12, 12-23 and 24-35 months old groups. The total scores of P50, P25, P10 and P3 of KPCS (Chinese version) of infant parents in urban areas in China were 41, 38, 33, and 29 respectively. When the scores of parents were 34-37, 30-33, and ≤ 29, they were judged as mild, moderate, and severe lack of parenting confidence. There was no significant difference in the Chinese version of KPCS between parents of different age groups and parents of different gender (χ²=3.53, P=0.171; χ²=1.41, P=0.236). Each factor score≤P3 is defined as the boundary score, and the corresponding boundary scores of "parenting" "support" and "competence" were 13, 9, and 5 respectively. Conclusion: The Chinese version of KPCS can be used to assess the parenting confidence of infants in urban areas of China. It can used as one of the bases for scientific and objective evaluation of the parenting status of families.

[Value of abnormal expression of Krüppel-like zinc-finger protein transcription factor 5 in the diagnosis and prognosis of liver cancer].

Objective: To explore the value of Krüppel-like factor 5 (KLF5), a family member of the zinc finger protein transcription factor, in the diagnosis and prognostic evaluation of hepatocellular carcinoma (HCC). Methods: Cancerous and non-cancerous tissues were collected from 126 cases after HCC surgery by self-matching method with microarray fabrication. Immunohistochemistry was used to analyze the expression of KLF5, clinicopathological characteristics and prognostic value. The sera of 222 cases with chronic liver disease were collected and their KLF5 levels were quantitatively determined by enzyme-linked immunosorbent assay (ELISA). Simultaneously, 40 normal human sera were used as controls to evaluate the value of abnormal KLF5 in the diagnosis and differentiation of benign and malignant liver diseases. T-test, Z-test and χ (2) test were performed on the data. Results: The positive expression rate of KLF5 in the HCC group was 95.2% (120/126), which was significantly higher than the non-cancerous group 38.9% (49/126; χ (2) = 14.385, P < 0.001). KLF5 expression was significantly correlated with TNM stage (stage I 35%, stage II 40%, stage III 74.4%, stage IV 78.1%), tumor size, alpha fetoprotein (AFP) concentration, portal vein embolism, HBV infection and 5-year survival rate. Univariate/multivariate analysis showed that KLF5 high expression was an independent predictor of HCC prognosis. The serum KLF5 level was significantly higher in HCC patients than liver cirrhosis, chronic hepatitis and normal control group (P < 0.001). With the serum KLF5 > 800 ng/ml and AFP > 25 µg/L as limit, the positive rates for HCC diagnosis were 90.48% and 73.81%, respectively, which were lower than the AFP specificity and false positive rate, and was helpful for the differential diagnosis of benign and malignant liver diseases. Conclusion: The overexpression of KLF5 in liver cancer tissues and blood is closely related to the HCC clinical stage and prognosis. Moreover, KLF5 analysis is helpful for HCC diagnosis and differential diagnosis.

[Abnormal expression of CD44 aggravates liver disease progression in patients with non-alcoholic fatty liver disease accompanied with hepatitis B virus replication].

Objective: To analyze the expression of CD44 in non-alcoholic fatty liver disease (NAFLD) accompanied with hepatitis B virus (HBV) infection and its clinical significance. Methods: Blood sample of hospitalized patients with NAFLD, chronic hepatitis B, cirrhosis, and healthy population (control) was collected. The study was approved by the hospital ethics committee. Serum CD44 level and clinopathological characteristics were analyzed quantitatively by enzyme-linked immunosorbent-assay. Flow cytometry was used to analyze the proportion of CD44(+)T lymphocytes in patients with NAFLD and chronic hepatitis B. NAFLD model was prepared with high-fat diet to verify the abnormal expression of CD44. Results: Compared with the healthy control group, the expression of serum CD44 in the cirrhosis group, chronic hepatitis B group and NAFLD group was increased, and the difference between the groups were statistically significant (P < 0.01). NAFLD patients graded as mild or severe group were equally accompanied by hepatocyte injury, abnormal blood glucose, lipid or CD44. In NAFLD patients accompanied with HBV infection, serum CD44 concentrations were significantly higher in HBsAg, HBeAg and HBV DNA positive group than HBsAg, HBeAg and HBV DNA negative group (P < 0.01). The proportion of CD44(+)T lymphocytes in peripheral blood of NAFLD and chronic hepatitis B group were 78.2% ± 16.3% and 68.5% ± 20.9%, respectively, and both groups (NAFLD and chronic hepatitis B) were significantly higher than the healthy control group (46.5% ± 20.5%) (P < 0.05). The high-fat diet model confirmed that in rat liver tissues the CD44 was overexpressed with fat deposition accompanied with liver cell damage, especially remarkable in liver tissues containing carcinogens. Conclusion: The abnormal expression of CD44 in patients with NAFLD may be related to the malignant transformation of HBV-related liver disease.

[Expression of tuftelin protein and its clinical value in HBV-related liver cancer tissues].

Objective: To analyze the expression of tuftelin protein (TUFT1) and its clinical value in hepatocellular carcinoma (HCC)-related liver cancer tissues. Methods: The biological information data of TUFT1 mRNA expression in liver cancer and non-cancer tissues were analyzed from the TCGA and Oncomine database. After the approval of the ethics committee, the self-pairing method was used to collect the postoperative cancer and para-carcinoma tissues of 132 HCC cases hospitalized between January 2009 and December 2014. Tissue microarray and immunohistochemistry (IHC) were used to analyze the expression of TUFT1 in liver tissues. According to IHC staining, liver cancer was divided into high TUFT1 and low/no expression group. Combined with clinical data, the clinicopathological characteristics were statistically analyzed between and within the groups. The 5-year overall survival (OS) and disease-free survival (DFS) was analyzed by correlation analysis. Results: IHC staining showed that TUFT1 in cancer tissue was localized in the cytoplasm and cell membrane, and its positive expression rate was significantly higher in the liver cancer group (87.1%) than the para-carcinoma group (64.4%) (χ (2) = 18.563, P < 0.001). TUFT1 expression intensity in patients with liver cancer was significantly correlated with HBeAg positive (χ (2) = 4.080, P = 0.043), tumor size (χ (2) = 9.388, P = 0.002), vascular invasion (χ (2) = 14.885, P < 0.001), TNM stage (χ (2) = 13.516, P < 0.001) and ascites (χ (2) = 5.940, P = 0.015). TUFT1 high expression was negatively correlated with OS and DFS (P < 0.001). Conclusion: The overexpression of TUFT1 is closely related to HBV replication, vascular invasion and poor prognosis, and it is expected to become a useful marker for liver cancer diagnosis and prognosis.

[Forcasting the prevalence of myopia among students aged 6-18 years in China from 2021 to 2030].

Objective: To predict the prevalence of myopia among Chinese students aged 6-18 years under different intervention scenarios from 2021 to 2030. Methods: The multi-state Markov model was developed based on the transition process of study stages and myopia statuses. The development of myopia was simplified into two statuses: non-myopia and myopia. Students aged 6-18 years were also divided according to their study stages including senior kindergarten, primary school (from Grade 1 to 6), junior school (from Grade 1 to 3) and high school (from Grade 1 to 3). The parameters were extracted from the National Myopia Investigation in 2018 and published articles of cohort studies. The transition probability was applied to simulate the intervention scenarios, and sensitivity analysis was carried out. Results: The cumulative incidence of myopia among Chinese school-aged children and adolescents would increase consistently. It would be 91.3% (min to max: 83.7% to 96.7%) upon graduation from high school. Without any intervention, the myopia prevalence would increase to 61.8% (min to max: 55.4% to 69.5%) by 2030 among Chinese school-aged children and adolescents. And the myopia prevalence among students in primary schools, junior schools and high schools would be 45.6% (min to max: 40.2% to 54.3%), 81.3% (min to max: 72.6% to 91.0%) and 90.5% (min to max: 82.4% to 96.7%), respectively, all higher than the national target. If the interventions could achieve 70% of the desired effect, the myopia prevalence would be lower than the national target at each stage. Conclusions: Without effective interventions, the prevalence of myopia among students aged 6-18 years may keep increasing in the next ten years. If the interventions achieve the desired effect, the national target for myopia prevention and control could be reached. It is urgent to identify more effective interventions and call on the whole society to participate in the myopia prevention action to achieve the national goal by 2030. (Chin J Ophthalmol, 2021, 57: 261-267).

Oxidative DNA damage is concurrently repaired by base excision repair (BER) and apyrimidinic endonuclease 1 (APE1)-initiated nonhomologous end joining (NHEJ) in cortical neurons.

AIMS: Accumulating studies have suggested that base excision repair (BER) is the major repair pathway of oxidative DNA damage in neurons, and neurons are deficient in other DNA repair pathways, including nucleotide excision repair and homologous recombination repair. However, some studies have demonstrated that neurons could efficiently repair glutamate- and menadione-induced double-strand breaks (DSBs), suggesting that the DSB repair mechanisms might be implicated in neuronal health. In this study, we hypothesized that BER and nonhomologous end joining (NHEJ) work together to repair oxidative DNA damage in neurons. METHODS: Immunohistochemistry and confocal microscopy were employed to examine the colocalization of apyrimidinic endonuclease 1 (APE1), histone variant 2AX (γH2AX) and phosphorylated p53-binding protein (53BP1). APE1 inhibitor and shRNA were respectively applied to suppress APE1 activity and protein expression to determine the correlation of APE1 and DSB formation. The neutral comet assay was used to determine and quantitate the formation of DSB. RESULTS: Both γH2AX and 53BP1 were upregulated and colocalized with APE1 in the nuclei of rat cortical neurons subjected to menadione-induced oxidative insults. Phospho53BP1 foci were efficiently abolished, but γH2AX foci persisted following the suppression of APE1 activity. Comet assays demonstrated that the inhibition of APE1 decreased the DSB formation. CONCLUSIONS: Our results indicate that APE1 can engage the NHEJ mechanism in the repair of oxidative DNA damage in neurons. These findings provide insights into the mechanisms underlying the efficient repair of oxidative DNA damage in neurons despite the high oxidative burden.

[Comparison the efficacy and prognosis of different first-line treatment for elderly diffuse large B-cell lymphoma].

Objective: To investigate the clinical features, survival and prognostic factors of elder patients with diffuse large B-cell lymphoma (DLBCL). Methods: The clinical data of elder patients with diffuse large B-cell lymphoma enrolled in the Cancer Hospital of Chinese Academy of Medical Sciences from April 2006 to December 2012 were retrospectively collected. All the patients were divided into R-CHOP-like group and CHOP-like group according to the dosage regimen. And the differences in demographic characteristics, clinical features, survival time and prognostic factors were compared between these two groups. Results: A total of 158 patients were enrolled, of which 78 patients in the R-CHOP-like group and 80 patients in the CHOP-like group were eligible. There were no significant differences between two groups on age, gender, pathological staging, B symptoms, bulky mass, ECOG score, IPI score, pathological type, LDH level, ß(2)-MG level, lymphocyte/monocyte ratio(LMR), neutrophils/lymphocyte ratio(NLR), platelet/lymphocyte ratio(PLR), Ki-67 index and bone marrow invasion. In the R-CHOP like group, the median progression-free survival (PFS) time was 10 months, and the median overall survival (OS) time was 30 months. The 1-year and 2-year PFS rates were 46.2% and 19.2%, respectively. The 1-, 2-, and 5-year OS rates were 79.5%, 59.0%, and 19.2%, respectively. In the CHOP-like group, the median PFS was 7 months, and the median OS was 15 months. The 1-year and 2-year PFS rates were 27.5% and 12.5% respectively. The 1-year, 2-year, and 5-year OS rates were 65.0%, 32.5% and 13.8%, respectively. The median PFS time and OS time in the R-CHOP group were significantly better than those in the CHOP group (P<0.05 for both). A stratified analysis showed that the PFS time and OS time were superior in the R-CHOP-like group compared to the CHOP-like group among patients older than 70 years (P<0.05 for both). In patients with stage Ⅲ-Ⅳ, the PFS time and OS time in the R-CHOP-like group were also superior to CHOP-like group (P<0.05 for both). Univariate Cox regression analysis showed that IPI score, LDH value, ß(2)-MG value, ECOG score, LMR, and PLR had an significant effect on prognosis (P<0.05 for all). Multivariate Cox regression analysis showed that lymphocyte/monocyte ratio and platelet/lymphocyte ratio were independent prognostic factors for diffuse large B-cell lymphoma (P<0.05 for both). Conclusions: The R-CHOP-like chemotherapy regimen is superior to the CHOP-like regimen in the first-line treatment of patients with diffuse large B-cell lymphoma. ECOG score, LMR and PLR may be independent prognostic factors for diffuse large B-cell lymphoma. ECOG score, LMR and PLR are independent prognostic factors.

[The outcome and safety of neoadjuvant PD-1 blockade plus chemotherapy in stage â ¡~â ¢ non-small cell lung cancer].

Objective: To explore the safety and therapeutic effect of programmed death 1 (PD-1) antibody combined with chemotherapy as a neoadjuvant therapy for patients with stage Ⅱ to Ⅲ non-small cell lung cancer (NSCLC). Methods: Thirteen patients, who had been diagnosed as stage Ⅱ-Ⅲ NSCLC and received PD-1 inhibitor plus chemotherapy as a neoadjuvant treatment in National Cancer Center/Cancer Hospital were recruited. The patients received consecutive neoadjuvant chemotherapy for 21 days as a cycle and the therapeutic efficacy was evaluated after two cycles. Results: At the last time of follow-up on December 2, 2019, the objective response rate (ORR) and disease control rate (DCR) of these patients were 61.5% (95% CI 30.9%-92.1%) and 100%, respectively. The downregulation rate of disease stage was 61.5% (8/13). The resectable rate was 38.5% (5/13), among them, the major pathologic response (MPR) was 60.0% (3/5) and the complete pathologic response (CPR) was 20.0% (1/5). The neoadjuvant chemotherapy displayed a low incidence of adverse reaction. The main grade 3 to 4 toxicities were neutropenia (38.5%) and leukopenia (23.1%). There was no significant immune-related toxicity. The safety and tolerability of perioperative period of patients underwent resection were promising. Conclusions: Immunotherapy combined with chemotherapy as a neoadjuvant treatment is an effective, low-toxicity treatment manner, which has perioperative safety and high rate of MPR for patients with resectable NSCLC. It is a promising treatment option for patients with stage Ⅱ to Ⅲ NSCLC.

[Phase â ¢ randomized controlled, multicenter, prospective study of recombinant anti-HER2 humanized monoclonal antibody (Cipterbin) combined with vinorelbine in patients with HER2 positive metastatic breast cancer: the HOPES Study].

Objective: To evaluate the clinical efficacy and safety of recombinant anti-HER2 humanized monoclonal antibody (Cipterbin) combined with vinorelbine in patients with HER2 positive metastatic breast cancer. Methods: Patients were randomized 2∶1 to test group and control group. Patients in test group received Cipterbin (4 mg/kg loading dose and 2 mg/kg maintenance dose each week, IV) combined with vinorelbine (25 mg/m(2) on days 1,8 and 15 of each 28 days, IV). Patients in control group received vinorelbine (25 mg/m(2) on days 1,8 and 15 of each 28 days, IV).The primary end point was progression free survival (PFS). Results: A total of 315 patients were enrolled from Jan 2009 to Jan 2013 (212 in test group and 103 in control group). The median PFS of test group was significantly longer than that of control group, 39.1 weeks vs 14.0 weeks (HR=0.24; 95%CI, 0.16-0.36; P<0.000 1). The objective response rate (ORR) and disease control rate (DCR) in test group were significantly higher than those in control group, ORR was 46.7% vs 18.45% (P<0.000 1) and DCR was 79.72% vs 45.63% (P<0.000 1). The incidence of neutropenia, leucopenia and erythrocytopenia were higher in both groups, but there was no significant difference between two groups.The most common adverse events associated with Cipterbin were infusion reactions. Left ventricular ejection fraction reduced to less than 50% in 5 patients, which were recovered. No serious cardiotoxicity. Conclusion: The recombinant anti-HER2 humanized monoclonal antibody (Cipterbin) combined with vinorelbine has significant efficacy and good safety. It is the optimized therapy regime for patients with taxane-pretreated HER2 positive metastatic breast cancer, which provides more targeted therapy opportunities for HER2 positive breast cancer patients in China.

[Intraobserver reproducibility of Ki-67 assessment of breast cancers based on digital slide].

Objective: To investigate the intra-observer reproducibility of Ki-67　　　assessment in breast cancers using three methods based on digital slide. Methods: Thirty cases of invasive breast cancer tissues were immunostained for Ki-67 by automatic stainer, and then scanned into digital pathological slides. Ki-67 positive index was measured individually by three pathologists using size-set visual assessment of hot spot (SSVAHS), size-set semi-automatic counting of hot spot(SSSACHS), and size-set automatic counting of hot spot (SSACHS), respectively, and repeated for 10 times. Intraclass correlation coefficient (ICC) of each assessment method was calculated, and the intraobserver reliability was classified as excellent, good, fair and poor according to ICC. Results: The ICC by 3 pathologists using SSVAHS was 0.832, 0.843 and 0.826, respectively, The ICC using SSSACHS was 0.926,0.938,0.929, and the ICC using SSACHS was 0.964, 0.971 and 0.968.The intraobserver reliability level of all three methods was excellent. Conclusion: The three methods of Ki-67 assessment achieve satisfactory intraobserver reproducibility, and the order of reproducibility from high to low is SSACHS, SSSACHS, and SSVAHS.

[Carcinoembryonic type specific markers and liver cancer immunotherapy].

Hepatocellular carcinoma (HCC) is a chronic inflammation derived from the background of hepatitis B and C virus (HBV and HCV) infection, chemical intoxicants, or non-alcoholic fatty liver disease. Cancerous liver cells can express and secrete a variety of relatively specific markers, such as carcinoembryonic type of alpha-fetoprotein (AFP), phosphatidylinositol-3 (Glypican-3, GPC-3), Wnt/ß-Catenin key molecule of signaling pathway Wnt3a and liver cancer specific GGT-II (HS-GGT), etc. Clinical analysis of carcinoembryonic markers not only contributes to diagnosis and prognosis of HCC, but may also be the target of HCC immunotherapy with a promising prospect of development and application. This article reviews the latest valuable advances in carcinoembryonic type specific molecular markers and liver cancer immunotherapy.

[Hypoxia-inducible factor-1α mediates and regulates angiogenesis-related factors expression in hepatocellular carcinoma].

Objective: To investigate the expression of hypoxia-inducible factor (HIF)-1α, vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) during the progression of liver diseases and the molecular mechanism of HIF-1α in regulating the expression of angiogenic factors in hepatocellular carcinoma (HCC). Methods: Serum samples from hospitalized patients with liver cancer, liver cirrhosis, and chronic hepatitis were collected, and healthy people were used as controls. Mouse models of hepatocarcinogenesis were used to detect the dynamic expression of HIF-1α, VEGF and Ang-2. Enzyme-linked immunosorbent assay was used to quantitatively analyze the serum levels of HIF-1α, VEGF and Ang-2 in patients with liver disease and mice. HIF-1α-specific miRNA expression plasmids was constructed to transfect HepG2 human HCC cells. HIF-1α mRNA transcriptional interference effects were analyzed on biological behavior, VEGF and Ang-2 expression, and epithelial mesenchymal transformation (EMT) in human HCC cell line. The sample means of multiple groups were compared by analysis of variance and q test and the sample rate was compared by χ (2) test. Results: In patients with chronic liver disease, the serum expression of HIF-1α, VEGF and Ang-2 in the liver cancer group (145.6 ± 32.6) µg/L, (458.9 ± 125.3) µg/L and (42.9 ± 5.1) µg/L was significantly higher than the liver cirrhosis (P < 0.001) (79.5 ± 28.4) µg/L, (206.8 ± 56.8) µg/L and (26.2 ± 6.1) µg/L and chronic hepatitis group (60.1 ± 18.8) µg/L, (178.1 ± 85.4) µg/L and (21.8 ± 6.9) µg/L. In addition, HIF-1α was positively correlated with VEGF (r = 0.937, P < 0.001), HIF-1α and Ang-2 (r = 0.933, P < 0.001), and VEGF and Ang-2 (r = 0.910, P < 0.001). Mouse models of hepatocarcinogenesis confirmed that HIF-1α, VEGF and Ang-2 had progressively increased during the process of malignant transformation from normal hepatocytes, hepatocyte degeneration, and precancerous lesions to canceration. HIF-1α miRNA intervention plasmid had transformed HepG2 cells. Compared with the blank group, HIF-1α mRNA, HIF-1α, VEGF and Ang-2 were decreased by 88.1%, 59.8%, 54.0% and 36.0% at 72h, respectively. The expression level of EMT-related protein Snail (0.26 ± 0.02 and 0.67 ± 0.09, q = 6.75, P < 0.003), VIM (0.27±0.08 and 0.73±0.04, t = 10.35, P < 0.001) and Twist (0.24 ± 0.07 and 0.73 ± 0.02, q = 12.08, P < 0.001) was significantly reduced, but the expression level of E-cadherin (0.76 ± 0.08 and 0.27 ± 0.09, q = 7.05, P < 0.002) was significantly increased. Conclusion: HIF-1α mediates and regulates angiogenesis-related factors such as VEGF and Ang-2 expression in hepatocellular carcinoma. Furthermore, HIF-1α transcriptional interference can significantly affect the biological characteristics and EMT transformation of hepatocellular carcinoma cells.

Chemical mapping of a single molecule by plasmon-enhanced Raman scattering.

Visualizing individual molecules with chemical recognition is a longstanding target in catalysis, molecular nanotechnology and biotechnology. Molecular vibrations provide a valuable 'fingerprint' for such identification. Vibrational spectroscopy based on tip-enhanced Raman scattering allows us to access the spectral signals of molecular species very efficiently via the strong localized plasmonic fields produced at the tip apex. However, the best spatial resolution of the tip-enhanced Raman scattering imaging is still limited to 3-15 nanometres, which is not adequate for resolving a single molecule chemically. Here we demonstrate Raman spectral imaging with spatial resolution below one nanometre, resolving the inner structure and surface configuration of a single molecule. This is achieved by spectrally matching the resonance of the nanocavity plasmon to the molecular vibronic transitions, particularly the downward transition responsible for the emission of Raman photons. This matching is made possible by the extremely precise tuning capability provided by scanning tunnelling microscopy. Experimental evidence suggests that the highly confined and broadband nature of the nanocavity plasmon field in the tunnelling gap is essential for ultrahigh-resolution imaging through the generation of an efficient double-resonance enhancement for both Raman excitation and Raman emission. Our technique not only allows for chemical imaging at the single-molecule level, but also offers a new way to study the optical processes and photochemistry of a single molecule.

[Effect of CHL1 gene on cell viability, invasiveness and apoptosis in neuroblastoma cells].

Objective: To investigate the effect and mechanisms of CHL1 gene overexpression on cell viability, invasiveness and apoptosis in neuroblastoma cells. Methods: The empty plasmid (pcDNA3.1 group) and CHL1 recombinant plasmid (pcDNA3.1-CHL1 group) were transfected into SK-N-SH human neuroblastoma cells, and the untransfected cells were used as blank control. Forty-eight hours after transfection, the protein expressions of CHL1, PCNA, MMP-2, Bax, STAT3 and p-STAT3 were detected by western blot. Meanwhile, cell viability, invasion and apoptosis were detected by MTT, transwell and flow cytometry assays, respectively. Results: The expression level of CHL1 protein in pcDNA3.1-CHL1 group was 0.612±0.052, which was higher than that of pcDNA3.1 group 0.122±0.014 and blank control group 0.120±0.013, with statistically significant difference (P<0.05). After 24, 48 and 72 hours of transfection, the absorbance (A) values of SK-N-SH cells in the pcDNA3.1-CHL1 group were 0.328±0.035, 0.502±0.051 and 0.688±0.064, respectively, whereas those in the pcDNA3.1 group were 0.562±0.050, 0.796±0.065 and 0.973±0.077, respectively. The differences were statistically significant (P<0.05). The invaded cells in the pcDNA3.1-CHL1 group were 104.9±3.7, which were lower than that in the pcDNA3.1 group (175.6±4.6), with statistically significant difference (P<0.05). Additionally, the apoptotic rate of pcDNA3.1-CHL1 cells was (23.46±1.22)%, which was higher than that in pcDNA3.1 group (3.45±0.20)%(P<0.05). Furthermore, the levels of PCNA, MMP-2, Bax and p-STAT3 proteins in pcDNA3.1-CHL1 group were 0.156±0.018, 0.122±0.015, 0.285±0.032 and 0.023±0.004, respectively, whereas those in pcDNA3.1 group were 0.542±0.053, 0.196±0.021, 0.073±0.009 and 0.057±0.007, respectively. There were statistically significant differences between two groups (P<0.05). Conclusion: Overexpression of CHL1 inhibits the cell viability and invasion, as well as induces apoptosis of neuroblastoma cells, which is related to the inhibition of STAT3 signaling pathway.