?Corneal collagen cross-linking ( CXL ) could increase the mechanical strength, biological stability and halt ectasia progression due to covalent bond formed by photochemical reaction between ultraviolet - A and emulsion of riboflavin between collagen fibers in corneal stroma. Corneal melting is an autoimmune related noninfectious corneal ulcer. The mechanism of corneal melting, major treatment, the basic fundamental of ultraviolet- A riboflavin induced CXL and the clinical researches status and experiment in CXL were summarized in the study.

Objective To construct recombinant lentivirus silence vector aiming at rictor gene in mTORC2 specific protein, and to investigate its regulation on mTORC2/SGK1 signal pathway and the effect on pulmonary alveolar epithelial sodium ion chan-nel,as well as the role in acute respiratory distress syndrome(ARDS)and acute lung injury.Methods The interfering vector plas-mid and empty vector plasmid of target gene rictor were constructed,which and the lentivirus packaging system were co-transfected to 293T cells.The viral supernatant was collected,centrifuged,concentrated and purified for obtaining recombinant lentivirus.The virus titer was detected and the virus was infected to A549 cells.Stable cell lines were screened.RT-PCR was used to confirm the silencing situation of target gene rictor.The expression situation of various signal indexes in this pathway was detected by PCR and Western blot.Results The recombinant lentivirus of silence gene rictor was successfully constructed and transfected to A549 cell for obtaining stable cell lines.Compared with blank and control groups,the mRNA levels of rictor,downstream SGK1 andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased (P <0.05 ).Meanwhile,the protein levels of rictor,downstream SGK1,P-SGK andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased compared with the other two groups(P<0.05).Conclusion Silence rictor gene has the obvious regulation effect on mTORC2/SGK1 signal pathway,meanwhile affects the expression of pulmonary alveolar epithelial cellular α-,β-and γ-ENaC at gene and protein level.It is speculated that mTORC2/SGK1 may be an important signal pathway for regulating the clearance capacity of pulmonary alveolar epithelial cells on pulmonary alveolar fluid and simultaneously affecting the pulmonary edema formation.

Objective To study the feasibility of transumbilical single-incision laparoscopic surgery ( SILS) in the treatment of tubal pregnancy, as compared with conventional multi-port laparoscopic surgery. Methods Sixty-two patients diagnosed as tubal pregnancy undergoing laparoscopic surgery at this hospital between September 2013 and June 2014 were selected for investigation. There were 29 cases of transumbilical SILS ( SILS group) and 33 cases of conventional multi-port laparoscopic surgery ( MPLS group) . We retrospectively reviewed the medical records of all patients and analyzed the surgical outcomes, including operative time, blood loss, surgical complications and hospital stay. Results In both groups, all procedures were performed without failure.No conversion to open surgery or additional skin incision was needed.There were no differences between the SILS and MPLS groups in the operation time [(51.5 ±10.8) min vs.(47.3 ±9.4) min, t=1.637, P=0.107], the mean estimated blood loss [(15.5 ±10.5) ml vs.(18.4 ±12.2) ml, t=-0.996, P=0.323], the absolute decrease of hemoglobin from preoperative to postoperative [(14 ± 5) g/L vs.(13 ±4) g/L, t=0.874, P=0.386], the postoperative hospital stay [(3.5 ±0.9) d vs.(3.8 ±0.8) d, t=-1.390, P=0.170], and the incidence of postoperative fever (2 cases vs.3 cases, χ2 =0.000, P=1.000).Follow-up for 1-3 months ( mean, 1.8 months) in all the cases showed smooth recovery and no postoperative complications. Conclusion SILS is a feasible and safe approach in the treatment of tubal pregnancy.

AIM To investigate antitumor effects of curcumin (Cur) combining with adriamycin(ADM) on human tumor cell lines in vitro . METHODS The antitumor effects of the drugs on tumor cell lines in vitro were determined with MTT method. The Jin's formula was used to analyse the effect of drug combination. RESULTS In simultaneous adiministration Cur 2 04 ?mol?L -1 ～16 29 ?mol?L -1 combining with ADM 0 70 ?mol?L -1 ～5 52 ?mol?L -1 produced a simple addition or potentiation effect. In sequential adiministration the first adiministration of ADM followed by Cur resulted in an antagonistic effect, while the change of the order of adiministration produced an simple addition effect. CONCLUSION Simultaneous adiministration of Cur and ADM produces synergistic effect but sequential adimistration of the drugs only produce a unidirectional synergistic effect.

Garcinol, a benzenetriol compound extracted from Garcinia cambogia, has antitumor activity, however, its antitumor mechanism remains unclear. The aim of this study was to investigate the role and mechanism of garcinol as a novel potential proteasome inhibitor. We applied the drug affinity responsive target stability (DARTS) method coupled to mass spectrometry to determine the binding protein of garcinol; the proteasome activity assay was used to determine the effect of garcinol on its hydrolase activity; immunofluorescence and proximity ligation assay (PLA) were used to detect the effects of garcinol on ubiquitin and RPN6; and flow cytometry were used to determine the effects of garcinol on cell apoptosis; and the anti-cancer effect was studied in organoid models. The results showed that RPN6 was a direct binding protein of garcinol; garcinol inhibited the hydrolase activity of proteasome, and induced the accumulation and aggregation of ubiquitin protein, and its proteasomal inhibitory effect was dependent on RPN6; further studies showed that garcinol induced oligomerization of RPN6 and formation of granules in the nucleus; finally, it was verified that garcinol induced apoptosis of tumor cells, and inhibited the growth of organoids of Apc^min/+ small intestine mice. These results suggest that garcinol is a potential proteasome inhibitor, which inhibits proteasome activity by directly targeting RPN6 on proteasome 19S, which in turn induces cell apoptosis and inhibits tumor growth.

The expressions of estrogen receptor (ER) α,ERβ1,and ERβ2 in thyroid carcinoma,thyroid adenoma,and adjacent normal thyroid tissue were determined by RT-PCR and immunohistochemistry,and the relationship of expression levels of three ERs and the main clinical pathologic parameters were evaluated.The result showed that ERα mRNA expression in papillary thyroid carcinoma(PTC) tissues was significantly higher than adjacent normal tissue,while,ERβ2 mRNA expression in PTC tissues was significantly lower than normal tissue ( P＜0.05 ).The positive expression of ERβ1 and ERβ2 in the thyroid carcinoma was related with the histological variant of carcinomas; the ERβ2 expressions were related with the lymph node metastasis and TNM staging of the carcinomas.These results suggest that the decreased ERβ2 expression may induce proliferation and carcinogenesis of thyroid follicular epithelial cells as well as progression of the carcinoma,and is a powerful prognostic indicator.

Objective To compare the dosimetric parameters between the use of Tandem and Ring (TR;Nucletron#090.617) or Tandem and Ovoid (TO;Nucletron#189.730) applicators during three-dimensional (3D) high-dose rate (HDR) brachytherapy (BT) for cervical cancer.Methods The records of 40 cervical cancer (ⅡB-ⅣA) patients treated with 3D-image-guided HDR-BT were reviewed.Of these 40 patients, 20 were treated with the TO applicator, and 20 with the TR applicator.The D100% and V150% of the clinical target volume (CTV) and the D2 cc of organs at risk (OAR)(the rectum, bladder, and small intestine) during 3D-HDR-BT using TO and TR were compared using the independent sample t-test.ResultsOverall metrics:CTV volume:66.04±13.86 cm3(TR) vs.65.67±15.08 cm3(TO)(P=0.052);CTV D100:3.71±0.34 Gy (TR) vs.3.37±0.49 Gy (TO)(P=0.016);CTV V150%:0.54±0.02(TR) vs.0.56±0.04(TO)(P=0.034);rectum D2 cc:3.38±0.30 Gy (TR) vs.2.95±0.80 Gy (TO)P=0.037);bladder D2 cc:4.33±0.39 Gy (TR) vs.2.93±1.27 Gy (TO)(P=0.00);and small ntestine D2 cc:3.04±1.02 Gy (TR) vs.3.41±0.57 Gy (TO)(P=0.171).Conclusions TR has better CTV coverage than TO during 3D HDR brachytherapy for cervical cancer.In addition, D2 cc of the rectum and bladder were both igher with TR than with TO, though there is no significant dosimetric difference in the small intestine between the two applicators.Therefore, tumor location, extent of invasion, and vaginal conditions should be considered when selecting the suitable pplicator for the treatment of cervical cancer.

Objective To investigate the expression of microRNA-34a (miR-34a) and silent information regulator 1 (SIRT1) in human lens epithelial cells under H2O2-induced oxidative stress.Methods Different concentrations of H2O2 (0 μmol · L-1,100 μ mol· L-1,200 μmol · L-1,300 μmol · L-1,and 400 μmol · L-1) were used to stimulate SRA01/04 cells for 24 hours.Cell viability was measured using cell counting kit-8 (CCK-8) assay.Cell apoptosis was detected by flow cytometry.Expression levels of miR-34a/SIRT1 were measured by RT-PCR.Results CCK-8 assay showed that a certain concentration range of H2O2 had a proliferation inhibition on SRA01/04 cells.There was a dose response relationship between 100 μmol · L-1 and 400 μmol · L-1.Compared with 0 μmol · L-1 H2O2 group,the difference was statistically significant (all P ＜ 0.01).According to flow cytometry results,apoptotic rate of SRA01/04 cells in control group and H2O2(100-300 μmol · L-1) groups were (6.1 ± 1.2)％,(26.3 ± 1.8)％,(32.5 ± 2.2) ％,and (64.7 ± 5.3) ％.Compared with 0 μmol · L-1 H2 O2 group,the differences were statistically significant (all P ＜ 0.01).RT-PCR test results showed that the expression of miR-34a increased significantly in a dose-dependent manner after the SRA01/04 cells treated with different concentrations of H2O2,while SIRT1 expression level was decreased,there were significant differences compared with control group (all P ＜ 0.001).Conclusion There is a significantly increase of miR-34a and decrease of SIRT1 in human lens epithelial cells under the oxidative stress of a certain concentration of H2O2.Down-regulated expression of miR-34a can increase the survival rate of human lens epithelial cells under H2O2-induced oxidative stress.

Objective To gain the virus like particles (VLPs) based on Trop2 targets ,which provided the basis for further in vi‐vo induced experiments and anti‐tumor vaccine studies .Methods Using molecular cloning method to constract eukaryotic expres‐sion vector of pCAGGs/Trop2 and baculovirus expression vector of pFastbac1/Trop2 ,expression product of pCAGGs/Trop2 in HeLa cells were intended to be to be anchored to the cell membrane by the methods of Immunohistochemistry ;pFastbac1/Trop2 were transformed into E .coliDH10bac isolates to gain recombinant bacmids ,which were transfected into insect cells to express re‐combinant baculovirus with Gag rBV ,then sucrose density gradient centrifugation ,Western Blot and electron microscope were per‐formed to purify and identify the Trop2 VLPs .Results Building recombinant bacmids which were transfected into insect cells to express recombinant baculovirus with Gag rBV ,gained the recombinant Virus like Particles .Conclusion Trop2 VLPs was success‐fully prepared ,which laid the foundation for the subsequent induction of humoral and cellular immune response .

Objective To study the clinical characteristics of lymphatic malformation(LM).Methods Clinical data of 14 cases with LM were analysed and literatures were reviewed.Results ①Among the 14 cases,6 were diagnosed as primary intestinal lymphangiectasia(PIL)and 8 as primary lymphedema(PL);②The major manifestations of PIL are symmetry edema,diarrhea,lymphocytopenia,hypoproteinemia and chylopericardium.The manifestations of PL included asymmetry edema of lower limbs and respiratory symptoms;③Directed oil-contrast lymphography and isotope techniques demonstrated absence of contrast in some lymphatic segments and blockage of chylous reflux;④White opaque spots were found on the duodenal and intestinal mucosa in the endoscopy of 6 cases of PIL indicating lacteal vessels dilation.Pathological examinations revealed intestinal lymphangiectasia;⑤Six cases of 14 patients were treated with surgical therapy and 8 cases were treated with conservative therapy including medium chain triglyceride(MCT).Conclusion LM should be diagnosed on the bases of typical clinical manifestations specific laboratory findings and by lymphography and pathology.Combination of surgical therapy and conservative therapy is recommended to treat the LM patients.