Objective To investigate the method of establishing lymph node metastasis model for gastric cancer in nude mice.Methods Human poorly differentiated gastric cancer SGC-7901 ceils were cul- tured,inherited,amplified and collected in vitro,and were inoculated in nude mice subcu.taneously.After pas- saging six generations,the subcutaneous tumor was orthotopically implanted in nude mice.The mice were then sacrificed on the 9th week after implantation.Serum CEA density was detected using radio-immunity method. Orthotopically implanted tumors,remote organ metastases as well as lymph node metastasis were observed macrescopically and histologically.Results The successful rate of orthotopically transplantion of gastric can- cer was 100 %.The rate of perigastric lymph node metastases was 93.3 %.Local infiltration and remote metastasis were observed.The level of serum CEA in nude mice bearing tumor was significantly higher than that of normal nude mice(P

OBJECTIVETo detect the expressions of receptor tyrosine kinases (RTKs) mRNA and protein and to explore potentially promising tumor markers and conceivable drug target in bladder cancer.

METHODSThe expressions of RTKs mRNA and protein in tissue from invasive urothelial carcinoma of the bladder were examined by real-time quantitative PCR array and cytokine antibody array, with normal bladder tissue as control. The Results were analyzed using bioinformatic approaches.

RESULTSThe expressions of TGFA, STAB1, SERPINE1, ANGPT2, SPINK5, ANGPTL1, PROK1, MDK, CXCL9, GRN, RUNX1, VEGFA, and TGFB1 were obviously upregulated in bladder cancer tissue, while those of EDIL3, PTN, CCL2, PDGFD, FGF13, KITLG, FGF2, SERPINF1, and TNF were downregulated. ALK, Btk, EphB2, ErbB4, PDGFR-α, ROS, Tie-2, Tyk2, and VEGFR3 were over-expressed in bladder cancer, while FRK, Fyn, IGF-IR, Insulin R, Itk, JAK1, JAK3, and LCK were low-expressed.

CONCLUSIONVascular endothelial growth factor/platelet-derived growth factor-targeted therapies may play an active role in treating carcinoma of bladder.

Carcinoma, Transitional Cell ; metabolism ; Humans ; RNA, Messenger ; genetics ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Urinary Bladder Neoplasms ; metabolism

OBJECTIVETo observe the change of Th immunological gene in renal transplant recipients after the treatment of cyclosporine (CsA) and tacrolimus (FK506).

METHODSThe peripheral blood lymphacytes just before and 24 hours after CsA and FK506 treatment were isolated. The total RNA of them were reverse-transcripted and examined by real-time quantity PCR array. The results were analyzed by bioinformatic methods.

RESULTSThe TLR4, CEBPB, IL4R, IL1R1,IL18R1,and IL1R2 genes were remarkably upregulated, whereas IL-2, CCL5, CD27, CCR5, CCR4, CD4, RPL13A, TGFB3, CD86, CCR3, STAT1, NFATC2IP, IL23A, IL15, IRF4, and TFCP2 were downregulated 24 hours after CsA treatment. The IL18, IL7, PTPRC, TNFSF4, SPP1, GFI1, TLR4, IL13RA1, TNF, INHBA, LAG3, IL13, IL1R1, SOCS5, IL10, YY1, TBX21, FASLG, IL18R1, and IL1R2 genes were remarkably upregulated, whereas IL-2, IL-3, IL-4, IL-6,CCR5, CD4, CD27, CD40LG, IL15, CCR3, CD86, CCR4, and IRF4 were obviously downregulated 24 hours after FK506 treatment.

CONCLUSIONCsA and FK506 exert their therapeutic effectiveness by regulating the expressions of a series of target genes.

Cyclosporine ; pharmacology ; Cytokines ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Kidney ; drug effects ; metabolism ; Kidney Transplantation ; Oligonucleotide Array Sequence Analysis ; T-Lymphocytes, Helper-Inducer ; drug effects ; metabolism ; Tacrolimus ; pharmacology

Background Retinal neovascularization disease is a common cause of blinding.Retinal neovascularization is related to enhancing proliferation of vascular endothelial cells.So how to inhibit proliferation of vascular endothelial cells is a hot burning issue.p21 is known to be involved in the regulation of cell cycle and therefore inhibit the cell proliferation.However,the relationship of p21 and the proliferation of vascular endothelial cells in retinal neovascularization disease is for further study.Objective The aim of this experiment was to study the proliferation of rhesus retinal vascular endothelial cells(RF/6A) and expression of p21 in RF/6A cells under the hypoxia condition,and discuss their association.Methods The RF/6A cells were cultured and passaged in vitro,then they were randomly divided into normoxia culture group(5％ CO2 +95％ O2) and hypoxia for 1 hour,3,6,12 hours group(1％ 02+5％ CO2 +94％ N2).Flow cytometer(FCM) was used to check the distribution of RF/6A cell cycle in the normoxia culture group and hypoxia for 1 hour,3,6,12 hours groups.MTT assay was used to detect and compare the cell proliferation(A570)among the various groups.The expression of p21 in the cells was analyzed by Western blot.Results FCM showed that the cells proportion of G0/G1 stage was reduced initially and then increased afterward in hypoxia for 1 hour and 3,6,12 hours groups,showing a significant difference among 5 groups (F =20.083,P =0.000),and the cells proportion of S stage and G2/M stage were increased firstly and then declined in different hypoxia groups with statistical significances (F =7.861,P =0.001 ; F =10.305,P =0.003).Compared with normoxia culture group,cells proportion of G0/G1 stage was declined and that of S stage and G2/M stage were raised after hypoxia culture,showing statistically signifcant differences(P＜0.05).MTT showed that cell multiplication capacity(A570 value)strengthened firstly and then weakened in hypoxia groups with time prolongation,showing a significant difference among all the groups(F=7.768,P=0.001),and A570 value in hypoxia for 3 hours and 6 hours groups (0.315± 0.062,0.365 ± 0.064) was significantly higher than that of the normoxia group (0.205 ± 0.063),respectively(P＜0.05).Western blot showed that the expression of p21 in the cells down-regulated at the beginning and then up-regulated with the increase of hypoxia time,and there was statistical significance (F =16.738,P=0.000).The p21 relative levels in different hypoxia groups were reduced in comparison with the normoxia group,showing statistical signifcances(P＜0.05).Conclusions Short-term hypoxia could reduce the expression of the p21 in RF/6A and induce cell proliferation initially,then p21 increases and cell proliferation is inhibited with the prolongation of hypoxia time.

A strain NK13 was screened for a certain extent asymmetric hydrolyze the rac-ketoprofen chloroethyl ester and identified as Bacillus megaterium. For the preparation of gene libraries, a positive clones was obtained from the tributyrin flat. The sequence of this esterase gene had been analysised, and contained the whole ORF of an esterase gene with the length of 933bp. The esterase gene of NK13 was compared with the esterase genes of GenBank and the result showed that the esterase gene of NK13 was a novel gene(GenBank accession nember DQ196347).The new esterase gene was inserted into the plasmid pET21b+, then the recombinant plasmid transformed E.coli BL21. After being induced by IPTG, it was expressed in the host strain. SDS-PAGE analysis showed that the relative molecular mass of the esterase was about 34kDa. The result of TLC and HPLC showed that the recombinant strain had higher conversion ration than templet strain. 47.4%of the rac-ketoprofen Chloroethl ester was hydrolyzed to ketoprofen by the recombinant strain in 45min. The (S)- ketoprofen enantiomeric excess, in the later,was 55.46%, which indicated that the esterase could hydrolyze (S)-ketoprofen chloroethyl ester firstly.

Objective To measure Ihe effect on rats spinal neuron flow according nerve roots repair time.Methods We adopted the experimental rats on the root avulsion and extravertebral foramen nerve root divison of C_(5～7).We divided them into four groupsin each which there were 16 ratsaccording the type of nerves root injury and repair timeGroup AC:the avulsed roots were reimplanted into the spinal cord and the transeeted roots were sutured to the proximal stump immediately.Group B,Dthe avulsed roots and the transected roots were reimplanted into the spinal cord or were sutured to the proximal stump in delayed 3 weeks each with 16 rats.At the different time point(3 weeks3 months6 months)through pathological examina- tion and immunohistological lechniques and nerve tracing techniqueswe examined the spinal cord and distal nerve trunk in order to observe the pathologic changes and axonal regeneration.Results Group A、C were much better than group B、D in the numberthe conformation and the degree of abatement of spinal motoneu- rons and nissl body.It is the same on the number and the development level of regenerating nerve fiber. Conclusion It had the advantage of neuronal protection and nerve regeneration that reparing the injured nerve roots earlv after nerve roots injury.

Boronic Acids ; therapeutic use ; Bortezomib ; Humans ; Male ; Middle Aged ; POEMS Syndrome ; therapy ; Peripheral Blood Stem Cell Transplantation ; Pyrazines ; therapeutic use ; Transplantation, Autologous

OBJECTIVETo compare the efficacy of different points combination in the treatment of menopausal insomnia.

METHODSNinety-six cases of menopausal insomnia were randomized into 3 groups, Xinshu (BL 15), Shenshu (BL 23), Sishencong (EX-HN 1), Shenmen (HT 7), Sanyinjiao (SP 6) were chosen in the restore interaction between the heart and the kidney group (group A, 32 cases); Zhaohai (KI 6), Jiaoxin (KI 8), Shenmai (BL 62), Pucan (BL 61) were chosen in the acupuncturing qiao mai group (group B, 32 cases); auricular Shenmen (TF4) and sensitive spot at the distribution area of auricular vagus nervus were chosen in the ear acupuncture group (group C, 32 cases). Six days made one session and the treatments were finished after 4 courses. The polysomnography (PSG) and Pittsburgh sleep quality index (PSQI) were employed before and after treatment to evaluate the alleviation of insomnia.

RESULTSThe parameters of the sleep latency (SL), rapid wave sleep latency (RL) and sleep efficiency (SE) were significantly improved in the three groups, and the differences were statistically significant (P < 0.05, P < 0.01). The SL and awaking time (AT) in group C [SL (401.08 +/- 16.54) min and AT (4.87 +/- 2.64) times] were significantly superior to those in the other two groups [SL (50.36 +/- 18.47) min, (54.87 +/- 20.92) min, AT (5.98 +/- 2.11) times, (6.13 +/- 3.04) times, all P < 0.05]. The S(3+4) (%) in group C was also significantly higher than those in the other two groups (both P < 0.05). It was indicated by PSQI that the sleep quality of group C (0.78 +/- 0.12) was significantly superior to that in group B (1.32 +/- 0.29), the total score and cured and markedly effective rate in group C [(4.34 +/- 1.43), 68.8% (22/32)] were superior to those in group A [(7.48 +/- 3.09), 53.1% (17/32), both P < 0.05].

CONCLUSIONEar acupuncture has a better curative effect than the restore interaction between the heart and the kidney group and acupuncturing qiao mai group, it is worth of being promoted.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Menopause ; psychology ; Middle Aged ; Sleep ; Sleep Initiation and Maintenance Disorders ; therapy ; Treatment Outcome

In this study, we use pot experiment to evaluate the effect of plant growth regulator on plant morphology and biomass allocation of Salvia miltiorrhiza. Different concentrations of uniconazole were supplied to S. miltioohiza by means of foliar spray. Height, breadth and stem diameter were measured dynamically, the biomass of leaf, stem, flower and fruit, root biomass and biomass ratio were also examined at the harvest time. Owing to the treatment, plant morphology showed significant changes, the height had been greatly reduced and the breadth decreased largely. Meanwhile, the biomass allocation changed too. The biomass ratio of leaf and stem had been notably reduced while the biomass ratio of root had been increased remarkably. It appears that foliar application of uniconazole during vigorous growth period in S. miltioohiza has dramatic effect on dwarfing plant and improving resistant to lodging. This measure could also be applied to condensed cultivation of S. miltioohiza to increase production.
Biomass ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; drug effects ; growth & development ; Plant Roots ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Salvia miltiorrhiza ; drug effects ; growth & development ; Triazoles ; pharmacology

AIM: To observe the efficacy of surgical excision combined with autologous limbus conjunctival flap transplantation in the treatment of pterygium accompanied with conjunctival cyst.METHODS: Totally 126 patients 188 eyes with pterygium were hospitalized in Department of Ophthalmology of Tongji Hospital of Huazhong University of Science and Technology during August 2013 and August 2015.The patients were divided into two groups: observation group (11 eyes of 11 patients) with pterygium accompanied with conjunctival cyst and control group (177 eyes of 115 patients) with primary pterygium.All patients underwent slit lamp microscope examination, anterior segment photography, and anterior segment optical coherence tomography(OCT).The size of pterygium was calculated by multiplying neck width and length of the covered corneal.All patients underwent excision combined with autologous conjunctival flap transplantation, and the resections were performed pathological section with hematoxylin and eosin staining.All patients were followed up postoperatively for 4-28mo.RESULTS: All cases in the observation group were confirmed by postoperative pathological examination.All cyst walls were complete, and containing single layer of epithelial cells.The mean size of pterygium of the observation group was 6.9±1.7mm2, and 6.3±1.8mm2 for the control group.There was no significant difference between the two groups (P>0.05).The mean postoperative healing time of observation group was 2.1±0.9d, and 1.9±0.8d for the control group.There was no significant difference between the two groups (P>0.05).Recurrence was seen in two eyes within the follow-up period in the control group, and no recurrence in the observation group.CONCLUSION: Surgical excision combined with autologous limbus conjunctival flap transplantation is a safe and effective treatment for pterygium accompanied with conjunctival cyst.