Objective To explore the influence of miR-375/SHOX2 axis in invasion and migration ability of esophageal squamous cell carcinomas （ESCC） by using cells in vitro， for searching potential new targets in therapy for ESCC. Methods Human ESCC cells，including kyse-70 and kyse-30 in the logarithmic growth phase were used in this study. The colony formation assay，Wound healing assay，MTT assay and Transwell assay were used to testify the effect of over-expression or down-expression of miR-375 or SHOX2 gene on ESCC cells. Then， rescue experiment was used to test the interaction between miR-375 and SHOX2. Results Compared with the negative control group， the proliferation rate， clone forming rate and penetrating number by Transwell were significantly reduced in cells after being over-expressed miR-375 or down-expressed SHOX2 （P＜0.01）. In contrast， compared with the negative control group， the proliferation rate， clone forming rate and penetrating number by Transwell were significantly increased in cells after being down-expressed miR-375 or over-expressed SHOX2 （P＜0.01）. SHOX2 expression was reduced by the over-expression of miR-375， while induced by the down-expression of miR-375 on ESCC cells. Rescue experiment showed co-transfection with miR-375 and SHOX2 reversed the low ability of proliferation and migration induced by over-expression of miR-375. Conclusion miR-375 could inhibit the ability of proliferation， invasion and metastasis of ESCC cells. SHOX2 could promote the ability of proliferation， invasion and metastasis of ESCC cells. The expression of SHOX2 could be regulated by miR-375 for proliferation， invasion and metastasis of ESCC cells. Therefore， miR-375 and SHOX2 could be new potential targets for drug treatment in ESCC.

Objective To investigate the effect of Rho kinase inhibitor Y27632 on cell proliferation，apoptosis and invasion and metastasis of non-small cell

lung cancer. Methods The non-small cell lung cancer cells A549 were treated with different concentrations of Y27632（5， 10， 20， 40， 80 μmol/L） and the

cells without treatment were set as the control group. The absorbance of A549 cells was detected by MTT method at 24，48，72 h after stimulation， and the

inhibitory rates of proliferation and half inhibitory concentrations（IC50）were calculated accordingly. The cell apoptotic rates were measured at 24 and 48 h by Annexin-enhanced green fluorescent protein （Annexin-EGFP）/iodinated propidium （PI） double staining via flow cytometry. The distribution of cell cycle， including G0/G1， S and G2/M， were detected by PI single staining with flow cytometry. The number of cells penetrating the membrane was detected by Transwell test. Results Y27632 can inhibit the proliferation of A549 （compared with the control group P＜0.05） in a dose- and time-dependence manner， and the IC50 value decreased with the prolongation of the time. Compared with the control group， the apoptotic rates and G0/G1 phase cell proportions were increased， and the proportions of S and G2/M cells were decreased， and the differences among different concentrations were statistically significant （P＜0.05）. Transwell experiments showed that Y27632 could inhibit the invasion of A549 cells. For instance， the number of cells penetrating the membrane were （78.1±6），（62.3±5.7），（51.7±5.2），（42.3±6.9），（36.4±5.3） and（22.8±4.6） for treatment with 0， 5， 10， 20， 40，80 mol/L Y27632， respectively. Conclusion In addition to the inhibition effect on cell invasion， Y27632 can inhibit the proliferation of A549 cells and induce apoptosis and G0/G1 arrest，having a certain value in the treatment of lung cancer.

bjective To explore the effects of morphine on the proliferation and apoptosis of A549 cells. Methods The CCK-8 was used to detect the

proliferation of A549 cells at 48 h after treatment with different concentrations of morphine （0.3，3，30 μg/ml）. Annexin-V FITC/PI double staining was used to detect the apoptosis rate of 30 μg/ml morphine at 48 h. Western blotting was used to detect the expression levels of X-linked apoptosis inhibitory protein （XIAP）， survivin， Bcl-2， caspase-3 and Bax of 30 μg/ml morphine at 48 h. Results Morphine could dose-dependently stimulate the proliferation of A549 cells at different concentrations， and the differences were statistically significant （P＜0.05）. Compared with the control group， the apoptotic rate of A549 cells was decreased after treatment with 30 μg/ml morphine （P＜0.05）. The treatment with morphine leaded to the increased levels of Survivin and XIAP but decreased level of caspase-3（P＜0.05）. Conclusion Morphine could promote proliferation and inhibit the apoptosis of A549 cells，possibly via the upregulation of XIAP and survivin and inhibition of caspase-3 activation.

Objective To investigate the radiosensitization effect of quercetin on cervical cancer HeLa cells and to explore its possible mechanism. Methods

Clone forming assay was used to observe the radiosensitizing effect of quercetin on HeLa cells after radiation with drug by various schedules and different drug concentrations（20% IC50 and IC50）. According to the experimental protocol， the experiments were carried out in radiation alone group， 20% IC50 quercetin+radiation group， radiation+20% IC50 quercetin group， IC50 quercetin+radiation group and radiation+IC50 quercetin group. The surviving fraction（SF） of above all groups after X-ray radiation of 12， 8， 6， 4， 2 and 0 Gy were calculated and the optimum schedule was selected. The below experiments were carried out in negative control group， quercetin group（20% IC50 and IC50）， radiation group and quercetin+irradiation group（20% IC50 and IC50）， and the dose of radiation was 4 Gy. Flow cytometry， DAPI staining， Western blotting（20%IC50 quercetin only） were explored to detect the cell cycle distribution， apoptotic changes and expression of Bcl-2 and Bax protein at 24 h after treatment. Results Compared with radiation alone group， the SF were reduced by the treatment of different concentration of radiation， quercetin in different sequence（P＜0.05）. The SF were inversely proportional to the concentration of quercetin， while the radiosensitization of quercetin was in concentration depended manner. Under the same concentration of quercetin and dose of X-ray，the SF in radiation+quercetin groups were lower than those in quercetin+radiation groups. DAPI staining showed that nucleus shrinkage were observed with the treatment of quercetin or radiation， and the combination of them could cause nucleus shrinkage and fragmentation more marked. Flow cytometry demonstrated that compare with other groups， quercetin+radiation groups could arrest cell cycle in G2/M phase（P＜0.05）. Western blotting showed that the levels of Bcl-2 protein was lowed in quercetin+radiation group than other groups（P＜0.05）；Bax levels increased in quercetin+radiation group， but it had no significance compared with quercetin group. Conclusion Quercetin has radiosensitization effect on HeLa cells and its effect is influenced by schedule and concentration of quercetin. The mechanism may be related to arrest of cell cycle in G2/M phase， cell apoptosis and the down-regulating the ratio of Bcl-2/Bax.

Objective To investigate the effect of speckle-type POZ protein （SPOP） on the biological behavior of bladder cancer T24by small interfering RNA （siRNA）. Methods The sequences of siRNA-1 and siRNA-1 together with LipofectamineTM 2000 were transfected into T24 cell lines to silence the expression of SPOP. T24 cells were divided into negative control group， siRNA1 and siRNA2 group. Quantitative real time polymerase chain reaction （qRT-PCR） and Western blotting method were used to detect the mRNA and protein levels of SPOP in each group at 48 h after transfection. Cell proliferation assay kit （CCK-8） was used to check the cell proliferation activity and the proliferation and migration were observed by Transwell migration assay and wound healing assay. Results The relative expression of SPOP mRNA were 1.012±0.025， 0.281±0.023 and 0.274±0.053 in control group， siRNA1 group and siRNA2 group. The relative expression of SPOP protein were 0.712±0.153，0.358±0.073 and 0.317±0.084 in control group， siRNA1 group and siRNA2 group. Both the mRNA and protein levels of SPOP were lower in siRNA1 group and siRNA2 group than control group（P＜0.05）. There was no significant difference in mRNA SPOP and protein levels between siRNA1 and siRNA2 groups （P＞0.05）. Compared with the control group，the proliferation activity of siRNA1 group and siRNA2 group were significantly increased，and the proliferation rates of siRNA2 and siRNA1 groups increased with the prolongation of transfection time（P＜0.05）. Results of Transwell migration test showed that the number of membrane penetrating cells were 128.6±9.3 and 134.5±8.6 in siRNA1 group and siRNA2 group， higher than 92.5±8.4 in control group with statistical significance （P＜0.05）. Results of the scratch test showed that the wound healing rates were （89.1±4.5）% and （93.7±5.1）% in siRNA1 group and siRNA2 group， higher than （32.6±2.8）% in control group with statistical significance（P＜0.05）.
Conclusion SPOP may play a tumor suppressor gene in the malignant behavior of bladder cancer via inhibiting proliferation and migration.

Objective To assess the efficacy and safety of anthracyclines in combination with taxanes for neo-adjuvant chemotherapy in patients with breast

cancer. Methods The data of 192 breast cancer patients treated by taxanes and anthracyclines neoadjuvant regimens was reviewed. The efficacy and safety of

taxanes and anthracyclines neoadjuvant regimens， as well as factors influencing pathological complete remission were analyzed. Results The efficacy of 192

patients could be evaluated. The effective rate was 84.9% （163/192），and the overall pathologic complete remission rate was 20.3%（39/192）. The complete

remission rates were 8.5%（10/118）in patients of HR（+）/HER-2（-）subtype， 42.8%（6/8）in patients of HR（-）/HER-2（+）subtype， 25.0%（5/20）in patents of HR（+）/HER-2（+） subtype， and 45.0%（18/40）in patients of HR-/HER-2-subtype. The logistic regression revealed that ER status affected the complete remission rate after neoadjuvant chemotherapy of the breast cancer. The main dose limiting toxicities was grade 3/4 neutropenia （94.7%）. Other non-hematologic toxicities included grade 2/3 vomitting （6.2%）， bone pain （14.1%）， febrile neutropenia （16.7%）， mucocitis （2.1%） and cardiotoxicity （5.7%） and etc. Conclusion Taxanes and anthracyclines chemotherapy regimens for the treament of patients with confirmed breast cancer show satisfied efficacy and safety， which can be regarded as an optimal neoadjuvant regimen for breast cancer.

Objective To observe the effects and adverse reactions of bevacizumab in brain metastases patients with serious cerebral edema. Methods

Retrospective analysis of 14 cases of brain metastasis patients with serious cerebral edema， including 7 cases of breast cancer， 5 cases of lung cancer， 1

case of esophagus cancer and 1 case of adenoid cystic carcinoma. Mannitol， dexamethasone and other conventional treatment were not effective in these patients. The symptom， quality of life and the volume of edema in T2 weighted MRI images before and after the bevacizumab treatment were compared. The adverse reactions were recorded in detail. Results The medium dose of bevacizumab was 4.76 mg/kg. The symptoms of 11 cases of brain metastases were significantly relieved after the bevacizumab treatment. Brain MRI indicated that the edema volumes before and after the treatment were reduced significantly［（38 804±14 859）mm3 vs.（80 100±28 338）mm3，P=0.02］. The edema index had a reduced trend（15.38±7.12 vs. 26.40±16.52，P＞0.05）. Adverse reactions associated with bevacizumab included bleeding in the nasal cavity and death in 1 patient， and hypertension in 3 patient. There were no proteinuria， anemia， stomatitis and other complications. Conclusion Bevacizumab has significant efficacy for treating serious cerebral edema， but patients should be selected carefully before receiving bevacizumab to avoid serious adverse reactions.

Objective To explore the clinical features， treatment and prognosis of primary pulmonary mucosa-associated lymphoid tissue lymphoma （MALTL）.

Methods From 2012 to 2015， seven primary pulmonary MALTL were treated in Nanjing Drum Tower Hospital. The clinical and follow-up data of seven patients were collected and their clinical features， treatment and prognosis were analyzed. Results Among seven patients of primary pulmonary MALTL， 2 cases were female and 5 cases were male with age between 45-62 years old. Six patients had respiratory symptoms and three of them had B-symptoms，with Ki-67 from 5% to 15% and International Prognostic Index （IPI） from 0 to 2. The PET/CT findings were nodules or consolidated mass with air bronchograms. Five patients received chemotherapy， including COP/CHOP with or without rituximab. The effect was partial remission or stable. Conclusion The symptom of primary pulmonary MALTL patients is nonspecific. PET/CT is useful in staging and evaluation of prognosis. The prognosis is overall good. Individual patient can appear certain invasive. The therapeutic consensus should be established with the clinical features， staging， IPI score and the extent of disease.

Objective To evaluate safety and efficacy of CT-guided percutaneous microwave ablation in pulmonary metastases from colorectal cancer. Methods From June 2010 to June 2015， 48 unresectable lesions in 32 patients with pulmonary metastases from colorectal cancer were selected for CT-guided microwave ablation. Kaplan-Meier method was used for survival analysis， and multivariable analysis of prognostic factors was done by Cox regression model. Results Among the 48 lesions of 32 patients， the complete response was achieved in 42 patients （87.5%）. No death was observed within 30 days after ablation， and no needle tract implantation metastases happened. Pneumothorax was found in 6 cases. All the patients were followed up. The median progression-free survival time was 26 months （95%CI： 16.790-35.210 months）. The 1， 2 and 3-year progression-free survival rates were 79.5%， 62.8% and 43.1%， respectively. Univariate analysis showed that tumor size， tumor-free interval and number of tumors were significantly related to progression-free survival time （P＜0.05）. Multivariate analysis showed that tumor size was an independent prognostic factor for progression-free survival （P＜0.05）.
Conclusion CT-guided percutaneous microwave ablation is a safe， effective and minimally invasive method to treat pulmonary metastases from colorectal cancer.

Objective To observe the clinical efficacy and adverse reactions of aprepitant in the prevention of nausea and vomiting induced by cisplatin chemotherapy. Methods One hundred and eight patients with lung cancer receiving cisplatin（25 mg/m2， d1-d3）-containing chemotherapy regimens in Zhejiang Cancer Hospital from August 1， 2014 to June 20， 2015 were enrolled. The antiemetic regiem for aprepitant group consisted of aprepitant（125 mg po d1，80 mg po d2-d3）， tropisetron（5 mg iv d1-d6）and dexamethasone（12 mg po d1，8 mg po d2-d4）. The regimen for control group was tropisetron（5 mg iv d1-d6） and dexamethasone（12 mg po d1，8 mg po d2-d4）. The primary endpoints were the percentage of patients with a complete remission（no nausea and no salvage treatment） during the acute phase（0-24 h） and the delayed phase（24-120 h）. The secondary endpoint was the percentage of patients with severe nausea during the entire study period（0-120 h）. Common Terminology Criteria for Adverse Events （CTCAE）4.0 was adopted for classification for adverse reactions. The functional living index-emesis （FLIE）was adopted to assess patients’ quality of life. Results The complete remission rates of vomiting for acute period in aprepitant group and control group were 90.7% and 75.9%（P=0.039）， respectively. The complete remission rates of vomiting for delayed period in aprepitant group and control group were 81.5% and 61.1%（P=0.019）， respectively. The aprepitant group’s vomiting preventive effect was better than the control group. The percentage of patients with severe nausea during the entire study period in aprepitant group and control group were 18.5% and 37.0%（P=0.032）， respectively. The FLIE score of aprepitant group and control group were 111.67±17.02 and 104.89±17.32（P=0.047）， respectively. The percentages of constipation and singultation of aprepitant group were higher than those of control group. Conclusion The aprepitant is effective for nausea and vomiting induced by cisplatin-containing chemotherapy. Further clinical study is warranted to assess the adverse reactions.

Objective To analyze the factors influencing the analgesic effect of pain in advanced cancer patients with moderate or severe chronic pain， and

provide some references to palliative care for advanced cancer patients. Methods Data were collected from 260 advanced cancer patients with moderate or severe chronic pain. All the patients received standard analgesic treatment. Pain intensity numerical rating scale（NRS）≤3， breakthrough pain≤3 times/day， salvage treatment number ≤3 times/day were used as the indicators of good pain control. Pain control after 3 days’ treatment， the analgesic time and analgesic doses needed for stable pain control were chosen as the evaluation indicators of analgesic complexity. The influences of following factors on the analgesic effects were analyzed， including gender， age， type of tumor， having bone metastases or not， pain intensity， pain location， pain causes， pain mechanism and having frequent breakthrough pain（more than 3 times a day） or not. Results In this study， 75.4% of the 260 patients had good pain control（NRS≤3） in 3 days， and the median analgesic time was two days. Logistic regression and Cox regression analyses showed that digestive system carcinoma， severe pain and frequent breakthrough pain were independent risk factors of poorer pain control in 3 days（P value were 0.032，＜0.001，＜0.001， respectively）， and also needed longer time for stable pain control（P value were 0.042， 0.002， 0.013， respectively）， while gender， age， bone metastases， pain location， pain causes and pain mechanism had no relationship with 3 days’ pain control and the time of analgesia（P＞0.05）. Of 185 patients who used strong opioids， patients younger than 60 years old（P=0.018）， or those with severe pain（P＜0.001）， neuropathic pain（P=0.002）， frequent breakthrough pain（P=0.015） needed more analgesic drugs， while gender， type of tumor， bone metastases， pain causes and pain location had nothing to do with the analgesic doses（P＞0.05）. Conclusion Younger than 60 years old， digestive system carcinoma， severe pain， neuropathic pain and frequent breakthrough pain were risk factors of pain control.

Objective To analyze the value of diagnosis and application of ultrasonography （USG）， contrast-enhanced CT and contrast-enhanced MRI on

metastatic neck lymph nodes in lingual squamous cell carcinoma. Methods Twenty-six patients with lingual squamous cell carcinoma confirmed by pathology were

enrolled in this study. The neck dissection zones were divided into 152 levels， and the metastatic lymph nodes were detected by USG， contrast-enhanced CT and contrast-enhanced MRI. The results were compared with pathologic data. Results Among the 152 levels， 83 positive lymph nodes were detected out of 1037 lymph nodes. Contrast-enhanced CT showed 28 positive levels， among which 15 were true positive； contrast-enhanced MRI detected 28 positive levels， among which 18 were true positive； USG showed 34 positive levels， and 24 levels were true positive. The sensitivity of contrast-enhanced CT， contrast-enhanced MRI and USG were 31.9%， 38.3% and 50.1%； the specificity were 87.6%， 90.5% and 90.5%； the accuracy were 70.4%，74.3% and 78.3%， respectively. Conclusion USG， contrast-enhanced CT and contrast-enhanced MRI are useful for the diagnosis of metastatic neck lymph nodes of lingual squamous cell carcinoma， and USG can be regarded as the best choice.

Objective To evaluate the efficacy of sorafenib in the treatment of hepatocellular carcinoma（HCC） through combined with peripheral circulating

tumor cell（CTC） and to improve the recognition of CTC in HCC. Methods CTCs of two HCC patients were isolated by SET-iFISH technique and were identified by i-FISH. Then we analyzed the patients’ clinical manifestation and long-term survival with CTC. Results CTCs of HCC were marked as CK-/DAPI+/FISH（CEP8 single）+/CD45-. We separated 2 patients’ peripheral blood （7.5 ml） in three consecutive days. Patient 1 had 2 CTCs，however， we had not found CTC in patient 2’ s blood. Till now， patient 2 have disease-free survival of 13 months. Conclusion Combination with CTCs can be an effective and noninvasive method to analyze the efficacy of sorafenib in the treatment of HCC. It can provide a new prospect in the diagnosis and treatment of HCC.

Collective invasion is one of the main movement styles of tumor cells. The roles of the collective migration of tumor cells in invasion and metastasis are attracting attention. Collective migration is the important movement pattern in the embryos development. Similarly， accumulating evidence suggests that the collective invasion is common style in malignancies. The potential mechanisms that collective invasion participates into spreading and metastasis of tumor cells are being revealed. The intensive research on collective cell invasion of cancer cells may be helpful to find the new therapeutic method for suppressing invasion and metastasis.

With increasing knowledge of the molecular subtypes and target therapy of colorectal cancer， more and more efforts have therefore focused on mitogen-activated protein kinase signaling （MAPK） pathway. V-raf murine sarcoma viral oncogene homolog B （Braf） is a principal downstream effector of MAPK pathway， and plays an important role in cancer development and progress. This article will review the role of Braf mutation in metastatic colorectal cancer， with emphasis on molecular and clinico-pathological characteristics， and current difficulties and progresses in treatment of colorectal cancer for reference.