Objective To establish a culture system in vitro of fetal and adult human retinal neural cells provide a model for the basic research of retinal neural cells and the medicinal exploitation. Methods Fetal human retinas(10~13 weeks after conception) and adult human retinas(20~40 years old) were dissected, dissociated, and put into culture plate which was coated with polylysine or rat tail gel. Specific growth factor EGF?FGF? BDNF or NT-4 were added to the culture medium. BrdU incorporation, Tunnel assessment and immuno-histochemistry and immuno-fluorescent staining were applied to determine cells proliferation, apoptosis and identify the component of cultured cells. Results Fetal human retinal cells and adult human retinal cells survived for up to 100 and 180 days in vitro. The addition of EGF?FGF? BDNF or NT-4 promoted the survival of both fetal and adult retinal neurons and stimultated proliferation of fetal retinal cells. The neurons or the rate of ganglion cells was observed with higher percentage in the group with growth factor adding than the group without. Conclusion Fetal and adult human retinal cells can be maintained in vitro and the fetal cells also can be expanded, which are helpful to generate retinal neurons for basic research and drug exploitation. The exogenous growth factors added to the culture medium can promote survival, proliferation and differentiation of retinal cells in culture.

OBJECTIVE:To evaluate efficacy and safety of caspofungin for non-effect/intolerant to fluconazole in patients with invasive fungal infetion(IFI) in intensive care unit(ICU).METHODS:A retrospective analysis was conducted in 14 IFI patients in ICU who was without effect/intolerant to gluconazole.They were treated with caspofungin in a dose of 70 mg in the first day,and then in a dose 50 mg?d-1 for 3～28 d.RESUITS:Of total 14 cases,2 cases had been diagnosed including 1 case of candidemia albicans and 1 case of pulmonary candidosis glabrata;5 probable cases including 2 cases of pulmonary aspergillus species,2 cases of pulmonary candidosis albicans and 1 cases of pulmonary candidosis parapsilosis;In 7 suspected cases pathogen fungus were not found out.One patient died of respiratory failure at third day of treatment.Clinical efficacy of death case cannot be evaluated.Of total evaluable 13 cases,2 were cured(2/13,15.4%),4 markedly effective cases(4/13,30.8%),2 improved cases(2/13,15.4%) and 5 ineffective cases(5/13,38.5%),the overall effective rate was 46.2%.Caspofungin can be tolerated by all patients during therapy without drug related adverse reaction.CONCLUSION:Caspofungin is certain of efficacy and safety for non-effect/intolerant to fluconazole in IFI patients in ICU,and is the first choice for IFI patients in ICU.

A new phenethanol, (2'R)-4-(2', 3'-dihydroxy-3'-methyl-butanoxy)-phenethanol (1), along with other eleven known benzene derivatives (2-12) were isolated from the roots, stems and leaves of Clausena excavata (Rutaceae). Compounds 3 and 4 are new natural products, and compounds 5-8, 10-12 were isolated from C. excavata for the first time. Their structures were elucidated on the basis of MS, 1D and 2D NMR spectroscopic analyses including HSQC, COSY and HMBC experiments. 1 was tested for its cytotoxicities against A549, HeLa and BGC-823 cancer cell lines, and antimicrobial activities against Candida albicans and Staphylococcus aureus. The results showed that 1 did not exhibit cytotoxic and antimicrobial activities.

Objective To discuss the clinical and pathological features of malignant rhabdoid tumor of the ureter (MRTU).Methods One case of MRTU was reported, a six-year-old girl was admitted to our hospital on May 29, 2014, and presented left loin pain 2 weeks, ultrasound showed gradually progressing hydronephrosis and hydroureter.During a physical examination, she felt tenderness in the left kidney area and no mass was palpable in abdomen.The ultrasound showed left sided gross hydroureteronephrosis and a round hyperechogenic mass in the inferior pole of the left ureter (In front of the left iliac vessel), with no obvious borders.Contrast-enhanced CT suggested a gross dilatation of the left kidney and ureter with a solidappearing lesion in the lower ureter;neither additional abdominal abnormalities nor enlarged lymph nodes were seen in both examinations.The surgery began with incision of left lower abdomen.The partial ureter of neoplasm was excised along with invaded psoas and posterior peritoneum by gross inspection, then ureteroureterostomy was performed.The severed ureter was completely blocked with the ill-defined neoplasm and was 3.3 cm in length and 2.1 cm in width.Results The ureteral neoplasm was excised,along with the invaded psoas and posterior peritoneum,after that ureteroureterostomy was performed.HE showed the diffuse large round nuclei, vesicular chromatin, prominent nucleoli cells, and moderate amounts of eccentrically placed eosinophilic cytoplasm.Immunohistochemical studies were positive for cytokeratin, epithelial membrance antigen and vimentin, negative for INI1, METU hereby was confirmed.She underwent a chemotherapy regimen consisting of ICE, alternating with VDC.Four courses chemotherapy (3 months) later,CT scan suggested hematogenous metastasis of lung.The family refused further treatment and the patient died of systemic metastasis eight months after surgery.Conclusion MRTU was a rare and highly aggressive tumor with a poor prognosis.

Objective To investigate the experience of diagnosis and management of coexisting ureteropelvic junction obstruction (UPJO) and nonreflux megaureter (NRM).Methods The retrospective study of UPJO with NRM was based on 10 years information retrieved from January 2005 to December 2015.The data of 13 patients (8 males and 5 females) were available and recorded.Mean age at surgery was 3.7 years old (range 1.8 to 14 years).The diagnosis and mangement were summarized.Coexisting ureterovesical junction obstruction (UVJO) and vesicoureteral reflux,iatrogenic stricture and vesicoureteral reflux were excluded.Intravenous pyelography,voiding cystourethrography,ultrasound and CT reconstruction were performed before operation.Only six patients had an accurate diagnosis as UPJO with UVJO before surgery.Pyeloplasty was the initial surgical management choice for 10 patients,and ureteroneocystostomy in 3 patients.Results UVJO were diagnosed with pyelography techniques in 3 patients after pyeloplasty,while 4 were diagnosed as nonreflux and nonobstruction megaureter.Of the 10 patients who underwent initial pyeloplasty,additional ureteroneocystostomy was required in 3 and the prognosis was good.Additional pyeloplasty was required in 2 of the 3 patients who initially underwent ureteroneoeystostomy.Mean follow-up time from last operation was 23.3 months (6-53 months),the overall prognosis was good.Conclusions It is often difficult to correctly diagnose coexisting UPJO and NRM.In patients with UPJO,it is highly recommended nephrostomy radiography after pyeloplasty to evaluate the distal ureterovesical junction.Initial pyeloplasty is always recommended as first-line therapy.Additional ureteroneocystostomy was required when hydroureteropelvic was aggravated.

BACKGROUND: In recent years, the incidence of inflammatory bowel disease is yearly increasing, and there are no effective treatment plans. Mesenchymal stem cells have achieved good outcomes in the treatment of immune-related diseases such as systemic lupus erythematosus, myocardial infarction and graft-versus-host disease. Numerous studies have shown that mesenchymal stem cells have their unique advantages in thetreatment of inflammatory bowel disease. Therefore, increasing concern has been placed on the treatment of inflammatory bowel disease by mesenchymal stem cells. OBJECTIVE: To summarize the existing research on the treatment of inflammatory bowel disease by mesenchymal stem cells, and to discuss its application and safety. METHODS: “Mesenchymal stem cells, inflammatory bowel disease, pathogenesis, treatment” were used as keywords to search CNKI, WanFang and PubMed databases for relevant literatures in the past 20 years both in Chinese and English. Finally, 62 papers were selected for review. RESULTS AND CONCLUSION: Mesenchymal stem cells have the abilities of self-renewal, tissue migration and pluripotency, as well as strong immunomodulatory effects. They not only show satisfying therapeutic effects in systemic lupus erythematosus, myocardial infarction, graft-versus-host disease and other immune-related diseases, but also have promising prospects in inflammatory bowel disease. However, little is reported in clinical studies, and there are still many problems, such as safety, encountered in the treatment of inflammatory bowel disease with mesenchymal stem cells.

This study is to investigate the protection effect of schisandrin B (Sch B) against oxidation stress of HK-2 cells induced by cisplatin and the mechanisms involved. HK-2 cells were cultured and divided into different groups: solvent control group, cisplatin exposure group, positive group, Sch B treatment group. Cell viability and toxicity were evaluated by MTT and LDH assay. GSH level and SOD enzymes activities were also measured. DCFH-DA as fluorescence probe was used to detect ROS level by fluorescence microplate reader. Nrf2 translocation was detected by Western blotting. Real time Q-PCR was used to detect expressions of NQO1, HO-1 and GCLC mRNA level. The results showed that Sch B could significantly inhibit the decline of cell viability induced by cisplatin treatment (P < 0.05) and the protective effect was in a dose dependent manner. Furthermore, Sch B treatment significantly inhibited the increase of ROS level induced by cisplatin and reversed the decrease of GSH level (P < 0.05). When Sch B concentration was up to 5 micromol x L(-1), SOD enzyme activities were also enhanced significantly compared with that of the cisplatin group (P < 0.05). It was shown that Sch B could cause nuclear accumulation of Nrf2 in association with downstream activation of Nrf2 mediated oxidative response genes such as GCLC, NQO1 and HO-1. These results suggested Sch B could protect against the oxidative damage of HK-2 cells induced by cisplatin via the activation of Nrf2/ARE signal pathway.

Objective To investigate the effect of COX￣2 inhibitor celecoxib on radiosensitity of irradiation￣resistant cell line CNE￣2R of nasopharyngeal carcinoma and the potential mechanism. Methods Via exposing to a series of X￣ray (2, 4, 6, 8 Gy, 3 times for each dose), radio￣resistant cell subline CNE￣2R was established from human nasopharyngeal carcinoma cell CNE￣2.Radiosensitivity was detected by clone formation assay.CNE￣2R and CNE￣2 cell lines were exposed to 25, 50, 75 μmol.L-1 celecoxib, respectively.Western blotting was used to detect the protein expression of COX￣2.Clone formation assay was performed to measure the survival fraction of CNE￣2 and CNE￣2R after radiotherapy alone or radiotherapy combined with 30 μmol.L-1 celecoxib treatment.Flow cytometry was used to measure influence of radiotherapy alone or radiotherapy combined with 30 μmol.L￣1celecoxib treatment on cell apoptosis.Number of residual γ￣H2AX foci was observed by immunofluorescence assay. Results The colony forming assay demonstrated that the values of SF2, D0 , Dq , and N of CNE￣2R cell subline [(0.81±0.05), (2.15±0.07) Gy, (2.94±0.08) Gy, (3.91±0.07), respectively] was significant higher than those of CNE￣2 cell line [(0.61±0.08), (1.47±0.06) Gy, (1. 68 ± 0. 10) Gy, (3. 13 ± 0. 05), respectively]. The expression of COX￣2 protein was significantly downregulated with increasing celecoxib concentration.Surviving fraction was decreased in both CNE￣2 and CNE￣2R cell lines after irradiation.After radiotherapy combined with celecoxib, apoptosis rates of CNE￣2 and CNE￣2R cell lines [(13.10±0.63)%, (5.30±0.75)%] were higher than those of the corresponding control groups [(4.90±0.71)%, (1.82±0.82)%].Celecoxib increased radiosensitivity in nasopharyngeal carcinoma CNE￣2R and CNE￣2 cell lines.The number of residual γ￣H2AX foci after irradiation was increased by celecoxib pretreatment.The difference was statistically significant (P<0.05). Conclusion Celecoxib can enhance radiosensitivity of radio￣resistant cell subline CNE￣2R of human nasopharyngeal carcinoma in vitro.

Aim To observe the effect of melittin on human hepatocelluar carcinoma HepG2 cell prolifera-tion in vitro and its further mechanisms.Methods The capacity of cellular proliferation and apoptosis was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,Hoechst 33258 assay and Annexin V-FITC /PI assay.The mR-NA expression of Shh, PTCH1, SMO, GLi1 and HDAC2 was performed by qRT-PCR.And the protein expression of Shh,PTCH1,SMO,GLi1 and HDAC2 was assessed by western blotting.Results Our study found that melittin effectively inhibited cell prolifera-tion and promoted cell apoptosis in vitro using MTT method and Flow cytometry.The mRNA and protein expression of Shh,PTCH1,SMO,GLi1 and HDAC2 were obviously decreased after treated with various con-centrations of melittin for 48h in HepG2 cells.Conclu-sions Taken together,our data suggest that melittin could inhibit cell proliferation and promote cell apopto-sis,reduce the level of HDAC2 and down-regulate the Hedgehog signaling pathway in this process simultane-ously.

Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.