The aim of this study was to construct antisense of ZNRD1 encoding gene, and to transfect it into SGC7901/VCR cells, to seek measures to overcome multidrug resistance of gastric cancer cells. ZNRD1 cDNA amplified by PCR was confirmed by DNA sequencing, and then inserted into the multiple cloning site of the expressing vector pcDNA3 1 + with molecular cloning technique. The recombinant vector was identified by endonuclease digestion. Antisense recombinant vector was transfected into SGC7901/VCR cells using lipofectamine. Flow cytometric analysis was used to detect adriamycin (ADM) accumulation in SGC7901/VCR cells. The results showed that a fragment was obtained by PCR, and its sequence was consistent with ZNRD1 cDNA reported in the literature. After antisense recombinant vector having been transfected into SGC7901/VCR cell, ADM concentration in such cells was increased. The above results indicated that the antisense vector ZNRD1 could enhance the intracellular accumulation of ADM in SGC7901/VCR cells, which might be of potential treatment value.

Objective To investigate the utility of plasma procalcitonin (PCT) as an early predictor for postoperative complications in patients who underwent elective pancreaticoduodenectomy (PD).Methods Clinical data of 87 patients who underwent elective PD in Changhai Hospital from March.1, 2016 to Dec.31, 2016 were collected.The general data, postoperative recovery, serum PCT level and white blood cell (WBC) count before, 1 d, 3 d and 5 d after PD were recorded.ROC curve was drawn and AUC was calculated to determine the cutoff value, sensitivity and specificity.Patients were divided into complication group (n=42) and noncomplication group (n=45) based on the occurrence of post-operative complications, and the comparisons between the two groups were performed.Results There were no significant differences on the age, gender, diabetes, obstructive jaundice, laboratory tests including PCT, operative time, blood loss volume during surgery and tumor type between the two groups, which were comparable.Complication group had longer hospitalization than noncomplication group (24 d vs 15 d,P<0.001), and the differences were statistically significant.In complication group, 18 patients had pancreatic fistula, 13 had peritoneal infection, 7 had gastric empty dysfunction, 8 had bleeding, 2 had bile fistula and 2 had incision infection after PD.The postoperative plasma PCT level in patients with gastric empty dysfunction, bleeding, bile fistula and incision infection was not statistically different from those in noncomplication group (all P>0.05), but the plasma PCT level in patients with pancreatic fistula and peritoneal infection on 3 d and 5 d after PD was significantly higher than those in noncomplication group, and the difference was statistically significant (all P<0.05).The combination of plasma PCT and WBC on 3 d and 5 d after PD was superior to PCT or WBC alone in predicting pancreatic fistula (sensitivity 88.9%, 72.7%;specificity 68.5%, 78.2%) and abdominal infection (sensitivity 100%, 100%;specificity 45.9%, 44.4%).Conclusions Plasma PCT could predict the occurrence of abdominal infection and pancreatic fistula after PD.The combination of PCT and WBC might be more valuable in predicting abdominal infection and pancreatic fistula.

Pancreatic cancer is the most lethal malignancy with an overall 5-year survival rate less than 9%, mainly due to late diagnosis and lack of effective therapeutic options.In the last decade, post-operative survival has been enhanced with advent of neoadjuvant therapy and combined adjuvant therapy.Furthermore, the information gained from the omics data, including next generation sequencing data, hasn′t yet begun to affect treatment of pancreatic cancer patients.However, in terms of precision medicine, pancreatic cancer has always lagged behind other tumors.Therefore, combined with practical experience, summary of the latest development and research progress of precise medical treatment of pancreatic cancer, especially from the fields of molecular biology and experimental models, is of critical importance. Further development of precise medicine for pancreatic cancer based on platforms using PDX and organoid model would promisingly help in effective improvement of clinical treatment.

Objective : To observe the clearance of smear layer on the root canal wall in different action time by scanning electron microscope (SEM), and to determine the optimal amount of time using sonically activated irrigation to wash root canal in clinic. Methods: Fifty-six ex vivo human anterior teeth with single straight root canal were selected. After routine mechanical preparation, they were divided into two experimental groups according to different irrigating agents: saline group and EDTA group. Each group was assisted by VDW sonic activation EDDY. The saline group was divided into three subgroups according to the irrigating time: 5 s, 30 s and 50 s; EDTA group was divided into six subgroups according to the irrigating time: 5 s, 10 s, 20 s, 30 s, 40 s and 50 s. The control group did not undergo root canal irrigation. After irrigation, the root was cut longitudinally. The smear layer of crown, middle and apical of root canal wall was observed by SEM. Results: After irrigating for 30 seconds, there was a significant difference between the normal saline group and the control group and the 5 second group (P<0.05), and there was no difference in the middle and apical part (P>0.05). After 50 seconds, there was a significant difference in the score of the smear layer between the apical area and the other groups (P<0.05). After irrigating for 5 seconds or 10 seconds in EDTA group, there was a significant difference between the scores of the crown and middle area of the root canal and the control group (P<0.05), and there was no significant difference in the apical area (P>0.05). There was a significant difference between the 20-40 second group and the first two groups (P<0.05). There was a significant difference between the 50 second group and the other groups (P<0.05). Comparing the cleaning effect on the smear layer after 50 seconds of irrigating between the two experimental groups, the whole root canal showed significant statistical difference (P<0.05). Conclusion The EDTA-assisted sonic activated device used for 50 seconds has the best cleaning effect.

Objective: To construct an aromatase-overexpressing breast cancer cell model and observe the real-time apoptosis of breast cancer cells induced by the aromatase inhibitor, letrozole. Methods: The lentivirus-mediated gene transfection method was used to construct the MCF-7-VC3AI and ZR7530-VC3AI cell lines,which stably expressed the apoptotic fluorescent indicator protein VC3AI.Simultaneously,letrozole-induced apoptosis models of the MCF-7-VC3AI and ZR7530-VC3AI breast cancer cell lines were also constructed.Real-time quantitative PCR(qPCR)and Western blot methods were used to detect the mRNA and protein levels of aroma-tase in the cells.Cell proliferation ability was measured using MTT.The proliferation of cells in vitro under testosterone,estradiol,or letrozole combined with testosterone treatments were also observed.Results:qPCR results showed that the expression of the aroma-tase mRNA was significantly higher in both the MCF-7 and ZR7530 cell models when compared to the MCF-7-VC3AI and ZR7530-VC3AI cell models.Western blot results showed that the expression of the aromatase protein was significantly increased in both cell models. MTT assay results showed that the proliferation of a cell model could be promoted by testosterone and estrogen stimulation.Under 100 nmol/L testosterone,the proliferation rate of over-expressed aromatase MCF-7-VC3AI cells was about 1.2 times than that of the control group(P<0.01)and the proliferation rate of ZR7530-VC3AI cells was about 1.5 times than that of the control group(P<0.01). However,letrozole inhibited the proliferation induced by testosterone in a dose-dependent manner.Under the effect of letrozole at 10 μmol/L,the proliferation rate of over-expressed aromatase MCF7-VC3AI cells was 80% of the control group(P<0.05),while the prolifer-ation rate of over-expressed aromatase ZR7530-VC3AI cells was 68% of the control group(P<0.05).Conclusions:The successful estab-lishment of cell models that can detect letrozole-induced apoptosis provides an important foundation for further investigating the mechanism of letrozole.

Objective The aim of this study was to investigate the distribution and liver-targeting properties of quercetin (QT)/coumarin 6 (C6)-loaded polylactic-co-glycolic acid-D-α-tocopherol polyethylene glycol 1000 succinate (PLGA-TPGS) nanoparticles (QCPTN) in a hepatocarcinoma ectopic transplantation solid tumor model using HCa-F cell-bearing mice.Methods The QT concentrations in biological samples were determined using reverse phase-high performance liquid chromatography (RP-HPLC) analysis.After intravenous administration to mice in the QCPTN and QTS groups,the QT concentration in plasma and in different tissues was simultaneously analyzed at the different time points.Detection indexes (relative targeting efficiency,Re;targeting efficiency,Te) and fluorescence inversion microscopy images of the frozen tissue (liver,solid tumor,spleen,lungs,kidneys,and heart) slices were used for quantitatively and qualitatively evaluating the liver-targeting properties of QCPTN in solid tumor-bearing mice.Results Te of the QCPTN group in the plasma,liver,solid tumor,spleen,lungs,kidneys,and heart were all greater than 3,indicating that the area under the concentration-time curve (AUC) of liver was more than three times that of the plasma and other organs.Fluorescence inversion microscopy images showed that the green fluorescence of QCPTN was mostly observed in the liver.Conclusion Using HCa-F cell-beating mice,QCPTN was found to have better in vivo liver-targeting properties in hepatocarcinoma ectopic transplantation solid tumor.

Pancreatic cancer is a common malignancy with the worst prognosis.Radical surgery has been the only curative treatment for pancreatic cancer.With the advancement of surgical techniques and the implementation of the concept of comprehensive treatment for cancer in recent years,neoadjuvant therapy for pancreatic cancer has received more attention.There are continuing controversies in the hotspots and difficulties,with opportunities and challenges coexisting.Four famous experts and their teams in pancreatic surgery discussed selection strategy of neoadjuvant therapy for pancreatic cancer based on clinical experiences.Professor Wang Chunyou proposed that surgery was prior for patients with a higher likelihood of achieving R0 resection for pancreatic cancer to avoid the possibility of tumor progression and loss the opportanity of radical resection during neoadjuvant therapy.For patients with less chance of radical resection for pancreatic cancer and unresectable pancreatic cancer,neoadjuvant therapy is worthy of a positive attempt.Professor Jin Gang and his team believed that neoadjuvant therapy played an important role in improving the survival time of patients with pancreatic head cancer,especially with borderline resectable pancreatic head cancer.After neoadjuvant therapy,pancreatic surgeons should pay attention to improvement of surgery safety and R0 resection rate.Professor Dai Menghua and his team suggested that patients with resectable pancreatic cancer and borderline resectable pancreatic cancer could benefit from neoadjuvant therapy,which required proof from clinical trials.Surgeons should choose the appropriate treatment strategy based on guidelines and individual conditions for patients with pancreatic cancer.Professor Shao Cheghao and his team suggested that surgical treatment after neoadjuvant therapy or translational therapy for locally advanced pancreatic head cancer is safe,effective and feasible,especially for pancreaticoduodenectomy with combined revascularization.For the treatment of patients with pancreatic head cancer after neoadjuvant chemotherapy,the choice of next treatment options,evaluation indicators,timing of surgery and surgical methods need to be further studied.

Objective: To compare the clinical therapeutic effects of arterial first approach pancreaticoduodenectomy(AFA-PD) with standard approach pancreaticoduodenectomy(SPD) in the treatment of borderline resectable pancreatic cancer (BRPC). Methods: A retrospective analysis of the clinical data of 113 cases of pancreatic cancer patients from January 2014 to August 2015 at Department of Hepato-Biliary-Pancreatic Surgery, Changhai Hospital, the Second Military Medical University, including 43 cases in AFA-PD group and 70 cases in SPD group.Every patient had gone high-resolusion computed tomography before the surgery, when BRPC was definitely diagnosed by both experienced radiologist and pancreatic surgeon.There were 24 males and 19 females in the AFA-PD group, with average age of (61.6±10.2)years.And in the SPD group, there were 47 males and 23 females, with average age of (62.7±9.4)years. Results: The operation time was (210.7±31.5)minutes in AFA-PD group, (187.9±27.4)minutes in SPD group, and peroperative bleeding volume was (1 007.1±566.3)ml in AFA-PD group, (700.0±390.0)ml in the other group.Those two indicators of AFA-PD group, compared with SPD group, were relatively higher, the difference was statistically significant(all P<0.01). And with regard to postoperative diarrhea(9.3% vs.5.7%), postoperative 1, 3 days of white blood cells(postoperative 1 day: (13.3±1.1)×10⁹/L vs.(12.4±2.4)×10⁹/L; postoperative 3 days: (12.7±1.6)×10⁹/L vs.(11.7±2.5)×10⁹/L), postoperative 1, 3, 5 days of peritoneal drainage fluid volume(postoperative 1 day: (184±42)ml vs.(156±54)ml; postoperative 3 days: (155±48)ml vs.(133±35)ml; postoperative 5 days: (66±20)ml vs.(47±31)ml), the differences between the two groups were statistically significant (all P<0.05). One patient in the SPD group was treated with unplanned secondary surgery for postoperative intraperitoneal hemorrhage, and the patient was cured and discharged.There was no death in the two groups within 30 days after surgical operation and no patient with positive gastric margin, duodenal margin, or anterior margin.The resection rate of superiormesenteric artery(SMA) margin R0 in AFA-PD group was higher than that in SPD group (P=0.019). The two groups were followed up for 14 to 30 months.As for AFA-PD group, the average survival time, progression free survival time and median survival time was respectively (20.4±1.2)months, (21.5±1.4)months and 20 months.There were 3 cases(7.0%) with local recurrence and 8 cases(18.6%) with liver metastasis or distant metastasis.In the SPD group, the average survival time, progression free survival time and median survival time was (17.1±1.1)months, (16.4±1.3)months and 16 months, respectively.There were 13 cases(18.6%) with local recurrence and 25 cases(35.7%) with liver metastasis or distant metastasis.As a result, the AFA-PD group had longer survival time(P=0.001)and progression free survival time(P=0.002). However, the lower local recurrence and distant metastasis rate in AFA-PD group did not reach statistical standard (P>0.05). Conclusion The arterial first approach pancreaticoduodenectomy is safe and effective in the treatment of borderline resectable pancreatic cancer, which can improve the resection rate of SMA margin R0, and prolong patient survival time.

Objective: To explore the role of nuclear factor-kappa B (NF-κB) signaling pathway in tamoxifen resistance of breast cancer cells. Methods: MTT assay was used to detect the inhibitory effect of 0-25 μmol/L tamoxifen on the proliferation of hormone recept or-positive breast cancer MCF7 and MCF7R/LCC9 cells. Real-time fluorescence quantitative PCR and Western blotting were used to detect the levels of NF-κB mRNA and its inhibitory subunit α (IKBα) protein in these two breast cancer cells, respectively. MCF7-GC3AI cells were infected with the lentivirus carrying the mutant plasmid pCDH-CMV-IKBα (S32A, S262A), pCDH-CMV-IKKβ (S177E, S181E) and the empty vector pCDH-CMV-Vector. Then the inhibitory effect of tamoxifen in combination with NF-κB signaling pathway inhibitor ACT001 on proliferation of NF-κB signaling pathway-inactivated, -activated and normal MCF7-GC3AI cells was detected by MTT assay. The apoptosis of MCF7-GC3AI cells in each group was observed under a fluorescence microscope, and the clone formation ability of these cells was detected by colony formation assay. Results: MCF7R/LCC9 cells were resistant to tamoxifen as compared with MCF7 cells (P < 0.01). The expression level of negative regulator IKBα of NF-κB signaling pathway in MCF7R/ LCC9 cells was decreased (P < 0.01), and the mRNA level of NF-κB was increased (P < 0.01) as compared with MCF7 cells. After treatment with tamoxifen for 48 h, the proliferation inhibitory ability and apoptosis rate of MCF7-GC3AI-IKKβ cells were decreased (P < 0.01, P < 0.05), and the cell colony formation ability was increased (P < 0.05) as compared with MCF7-GC3AI-Vector cells. But the proliferation inhibitory ability and apoptosis rate of MCF7-GC3AI-IKBα cells were increased (P < 0.05, P < 0.01), and the cell colony formation ability was decreased (P < 0.05) after tamoxifen treatment for 48 h. ACT001, an inhibitor of NF-κB signaling pathway could increase the cytotoxicity of tamoxifen in MCF7R/LCC9 cells (P < 0.01). Conclusion: NF-κB signaling pathway maybe play an important role in the development of tamoxifen resistance in hormone receptor-positive breast cancer cells.

This paper reports the diagnosis and treatment of a case of lymphocytic hypophysitis in pregnancy complicated by intrahepatic cholestasis of pregnancy. The 32-year-old patient had regular menstrual cycles and conceived naturally. Around 24 weeks of gestation, she developed a headache without abnormalities in the visual field. Skin itching occurred at 32 weeks of gestation, but her headache was significantly relieved. By 33 +1 weeks of pregnancy, the patient experienced elevated total bile acid, transaminase, and hypoglycemia, leading to a diagnosis of intrahepatic cholestasis of gestation (mild). Subsequently, at 33 +6 weeks of gestation, her total bile acid increased substantially, and the disease progressed rapidly into intrahepatic cholestasis of pregnancy (severe). As a result, an emergency cesarean section was performed to terminate the pregnancy. The Apgar score of the newborn was five points at one minute and ten points at five minutes. Following the surgery, the patient showed no lactation and had symptoms such as nausea, vomiting, fatigue, and refractory hyponatremia. Blood hormone measurement showed hypopituitarism. An enhanced intracranial MRI showed pituitary enlargement and uniform enhancement. Consequently, the clinical diagnosis was lymphocytic hypophysitis. Treatment with glucocorticoids effectively alleviated the symptoms. Menstruation resumed after more than one month of delivery. The intracranial MRI in postpartum reexamination after six months of delivery revealed a disappearance of the lesion. The patient continued with oral glucocorticoid therapy.