Background:Esophageal cancer is a common gastrointestinal cancer with poor prognosis,and effective chemotherapy is lacking currently. Studies have shown that cardiac glycosides can inhibit tumor cells growth,but its mechanism has not been fully clarified. Aims:To investigate the effect and mechanism of ouabain in regulating proliferation of human esophageal carcinoma cells. Methods:OE19 human esophageal carcinoma cells were treated with ouabain,and cells in control group were treated with DMSO. Cell proliferation was assessed by cell counting method. mRNA expressions of Sox2,Sox4,Sox7,Sox9 and Sox10 were determined by real-time PCR. Protein expression of Sox4 was determined by Western blotting. Gene expressions of phospho-histone3( ph3),a cell proliferation marker and Sox4 were detected by immunofluorescence staining. Results:Ouabain( ≥ 40 nmol/ L)could significantly inhibit OE19 cells proliferation. mRNA and protein expressions of Sox4 were significantly decreased in OE19 cells in ouabain(40 nmol/ L)group than those in control group(P < 0. 05). No significant differences in mRNA expressions of Sox2,Sox7,Sox9 and Sox10 were found between the two groups(P > 0. 05). Gene expressions of ph3 and Sox4 in nucleus of OE19 cells were decreased in ouabain (40 nmol/ L)group than those in control group. Conclusions:Ouabain is effective in inhibiting human esophageal carcinoma cells proliferation,the underlying mechanism might be related with down-regulation of Sox4 expression and the subsequent cell cycle modulation.

OBJECTIVETo assess the population genetic diversity and genetic structure and screen species-specific bands for identification of Changium smyrnioides and Chuanminshen violaceum.

METHODSeven wild populations of Changium smyrnioides and one cultivated population of Chuanminshen violaceum were studied by ISSR analysis. The population genetic diversity and population genetic structure were assessed by using POPGENE software.

RESULTA total of 152 ISSR markers were scored, among which 136 (90.8%) were polymorphic. The values of Gst tended to be high (mean Gst = 0.575). The level of genetic divesity of Changium smyrnioides (A = 1.272; P = 27.26%; I = 0.132; H = 0.087) was higher than that of Chuanminshen violaceum (A = 1.217; P = 21.7; I = 0.103; H = 0.067).

CONCLUSIONThe genetic variation of Changium smyrnioides is high and the majority of genetic variation occur among populations. Substantial genetic divergence is shown by cluster analysis (UPGMA) to befound between Changium smyrnioides and Chuanminshen violaceum at DNA level. In addition, one species-specific marker has been obtained in Chuanminshen violaceum. The phylogenetic relationship of two species has also been discussed.

Apiaceae ; classification ; genetics ; China ; Cluster Analysis ; DNA, Plant ; genetics ; Ecosystem ; Gene Frequency ; Genetic Markers ; Genetic Structures ; Phylogeny ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Repetitive Sequences, Nucleic Acid ; Species Specificity

AIM: To investigate the clinical characteristics of patients with systemic lupus erythematosus ( SLE) complicated with retinopathy. METHODS: Totally 121 cases of SLE patients treated in our hospital from February 2015 to February 2017 were selected,including 30 cases with retinopathy(observation group) and 91 cases without retinopathy(control group), the clinical manifestations and laboratory examination results of the two groups were compared. RESULTS: ln the observation group, there were 6 patients with bilateral retinopathy and 24 patients with monocular retinopathy. Cotton retinal exudation, retinal vascular occlusion and retinal hemorrhage lesions were common, accounted for 33%, 25% and 19%. The incidence of skin rash, skin vasculitis and Raynaud's phenomenon in the observation group were 63%, 47% and 37%, respectively, which were significantly higher than those in the control group (P<0.05). There were no significant difference in the incidence of mucosal ulcer, arthritis, neuropsychiatric symptoms and pleurisy between the observation group and the control group(P>0.05). The positive rate of anti ds-DNA in the observation group was 63%, which was significantly higher than that in the control group (P<0.05). The observation group and the control group urine protein > 3 +, anti Sm antibody positive and rRNP positive and antiphospholipid antibody percentage differences were not statistically significant(P>0.05). The SLEDAI score of the observation group was 20.14 (9, 30) points, which was significantly higher than that of the control group (P<0. 05). The modified BenEzra score of the observation group was 10 04士3.15,and was positively correlated with the SLEDAI score (rs=0.706,P<0.05). CONCLUSION: SLE with fundus retinopathy patients mainly manifest as cotton wool spots, retinal vascular occlusion and retinal hemorrhage; rashes, vasculitis, Raynaud's phenomenon,and anti ds-DNA antibodies are common in SLE patients with retinopathy. The modified BenEzra score plays an important role in the evaluation of SLE disease activity and retinal vascular lesions.

OBJECTIVETo investigate the effect of puerarin on the neural function and the histopathological changes after ischemic spinal cord injury in rabbits.

METHODSThirty male New Zealand white rabbits were randomly divided into three groups as follows: puerarin group (n=10) receiving intravenous infusion of 30 mg/kg puerarin for 10 minutes, control group (n=10) receiving intravenous infusion of the same volume of normal saline as puerarin for 10 minutes, and sham operation group (n=10) undergoing only the surgical exposure of the abdominal aorta. Temporary spinal cord ischemia was induced by infrarenal aortic occlusion for 20 minutes and followed by reperfusion. The neural status was scored with the Tarlov criteria at 8, 12, 24 and 48 hours after reperfusion. All the animals were killed at 48 hours after reperfusion and the spinal cords (L5) were removed immediately for histopathological study.

RESULTSThe neural function scores at 8, 12, 24 and 48 hours after reperfusion were higher in the puerarin group and sham operation group than those in the control group (P<0.05). More normal motor neurons in the anterior horn of spinal cord were present in the puerarin group and sham operation group than those in the control group (P<0.01). There was a strong correlation between the final neural function scores and the number of normal motor neurons in the anterior horn of spinal cord (r=0.839, P<0.01).

CONCLUSIONSPuerarin can significantly ameliorate the neural function and the histopathological damages after transient spinal cord ischemia in rabbits.

Animals ; Isoflavones ; pharmacology ; Male ; Motor Neurons ; pathology ; Rabbits ; Spinal Cord Ischemia ; drug therapy ; pathology ; physiopathology ; Vasodilator Agents ; pharmacology

Aromatase is a key enzyme responsible for in vivo estrogen biosynthesis. Inhibition of the activity of the aromatase has become an alterative way for treatment of breast cancer. In this review, the structure and catalytic mechanism of the aromatase is briefly introduced followed by thorough review of the progress in the study of the steroidal and non-steroidal aromatase inhibitors. This review is focused on the natural compounds that exhibit the aromatase inhibition, which include flavonoids, xanthones, coumarins, and sesquiterpenes. The structure-activity relationship of these compounds is also discussed.
Androstenedione ; analogs & derivatives ; Antineoplastic Agents ; chemistry ; pharmacology ; therapeutic use ; Aromatase ; chemistry ; metabolism ; pharmacology ; Aromatase Inhibitors ; chemistry ; classification ; pharmacology ; therapeutic use ; Breast Neoplasms ; drug therapy ; Catalysis ; Coumarins ; chemistry ; pharmacology ; Estrogens ; biosynthesis ; Flavonoids ; chemistry ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Nitriles ; chemistry ; pharmacology ; Sesquiterpenes ; chemistry ; pharmacology ; Structure-Activity Relationship ; Triazoles ; chemistry ; pharmacology ; Xanthones ; chemistry ; pharmacology

OBJECTIVE: To investigate the therapeutic effects of conventional treatment with different routes of administration of dexamethasone on sudden deafness. METHOD: Eighty-four patients with sudden deafness were included in this prospective randomized study. Twenty one patients (group 1) were treated with taking dexamethasone orally combined with conventional methods. Another 21 patients (group 2) were treated with intravenous dexamethasone injection combined with conventional methods. Group 3 (21 patients) were treated with intratympanic dexamethasone injection by the way of external ear combined with conventional methods. The other 21 patients (group 4) were treated with intratympanic dexamethasone injection by the way of pharyngotympanic tube combined with conventional methods. The hearing gains at 0.5, 1.0, 2.0, 4.0 kHz and the mean values were compared among four groups. RESULTS: The average hearing gains of 1, 2, 3 and 4 group was 21.3 dB, 27.5 dB, 43.2 dB and 48.1 dB respectively. Group 3 and group 4 had statistical difference compared with group 1 and group 2 in the average hearing gains. There was no obviously statistical difference between group 1 and group 2 and between group 3 and group 4. In patients with PTA < or = 70 dB, the average hearing gains at 0.5, 1.0, 2.0, 4.0 kHz had no obvious difference (P > 0.05) among four groups. However, in patients with PTA > 70 dB, there was statistical difference between group 1, 2 and group 3, 4 (P < 0.05), the hearing gains of group 3. 4 were apparently higher than that of group 1, 2. However, there was no significant difference of hearing gains between group 1 and group 2 (P > 0.05) and between group 3 and group 4 (P > 0.05). CONCLUSION The conventional drug treatment with taking dexamethasone orally or intravenous dexamethasone injection had no obvious effect on sudden deafness with PTA > 70 dB, but the conventional drug treatment with intratympanic dexamethasone injection is a useful treatment for sudden deafness. Comparison with whole body administration, intratympanic dexamethasone injection is more convenient to use in clinic, and with less prohibitions and complications. Patients with PTA > 70 dB should take intratympanic dexamethasone injection in early days.
Adult ; Dexamethasone ; administration & dosage ; therapeutic use ; Drug Administration Routes ; Female ; Hearing Loss, Sudden ; drug therapy ; Humans ; Male ; Middle Aged ; Prospective Studies ; Treatment Outcome

OBJECTIVETo construct the recombinant adenovirus containing herpes simplex virus-1 virion protein (VP) 22 and human microdystrophin gene, then the adenovirus was transfected into C2C12 myoblast and studied on the property of protein transduction with VP22-mediated microdystrophin in C2C12 myoblast.

METHODSThe full-length VP22 cDNA was obtained from recombinant plasmid pSINrep5-VP22 with PCR, and the product was directionally inserted into pShuttle-CMV to acquire the plasmid pCMV-VP22. Microdystrophin cDNA was obtained from recombinant plasmid pBSK-micro digested with restrictive endonuclease NotI, and the product was directionally inserted into pCMV-VP22 to acquire the plasmid pCMV-VP22-MICDYS. The plasmid of pCMV-VP22-MICDYS was lined with Pme I, and the fragment containing VP22-microdystrophin was reclaimed and transfected into E1 coli BJ5183 with plasmid pAdeasy-1. After having been screened by selected media, the extracted plasmid of positive bacteria was transfected into HEK293 cells with liposome and was identified by observing the cytopathic effect of cells and by PCR method to acquire the recombinant adenovirus Ad-VP22-MICDYS. Finally, the C2C12 myoblast were transfected with the recombinant adenovirus Ad-VP22-MICDYS and Ad-MICDYS, and the expression of microdystrophin was detected by RT-PCR, Western blot and immunocytochemistry.

RESULTSThe recombinant adenovirus including VP22 and microdystrophin gene was successfully constructed. VP22 transferred VP22-microdystrophin fused protein from infected C2C12 myoblast into uninfected cells and enhance the expression of microdystrophin in myoblast.

CONCLUSIONSRecombinant adenovirus containing VP22 and microdystrophin gene was constructed successfully. VP22 can enhance the expression with microdystrophin in myoblast. It lays the foundation for further studying on VP22-mediated recombinant including microdystrophin gene to cure Duchenne muscular dystrophy.

Adenoviridae ; genetics ; physiology ; Animals ; Cell Line ; Dystrophin ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Humans ; Mice ; Myoblasts ; metabolism ; virology ; Simplexvirus ; genetics ; metabolism ; Transduction, Genetic ; Viral Structural Proteins ; genetics ; metabolism ; Virion ; genetics ; metabolism

OBJECTIVEto investigate the effect of somatostatin on inflammatory immune disorders and prognosis in patients with severe sepsis caused by abdominal diseases.

METHODSfifty-three patients with severe abdominal sepsis (age > 18 years, APACHE-II score > 15) from June 2005 to June 2009 were randomly divided into Somatostatin group (n = 23) and SSC Group (n = 30). Fifteen healthy volunteers of the same age range were chosen as Control group. The SSC group was treated with classical SSC therapy, and the Somatostatin Group was treated with the same regime plus 14-peptide somatostatin continuous infusion at the dose of 6 mg/24 h for 7 days. The serum levels of interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) were determined by using ELISA. CD(4)(+), CD(8)(+) T cell subsets were determined by fluorescence activated cell sorter(FACS) and CD(4)(+)/CD(8)(+) was calculated. APACHE-II score was observed on admission (d1) and day 3, 7 and 14 after treatment. Morality rates in 28 days in two groups were recorded.

RESULTScompared with Control group, IL-10 and TNF-α levels were significantly elevated in patients with severe abdominal sepsis (P < 0.05), while CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) decreased significantly (P < 0.05). Compared with the Somatostatin group CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) on d7 and d14 in SSC Group were significantly increased (P < 0.05), while IL-10 and TNF-α decreased significantly(P < 0.05). APACHE-II scores on d3, d7, d14 of Somatostatin group were significantly lower than those of SSC group, and 28 d mortality rate also declined.

CONCLUSIONSin patients with severe abdominal sepsis, systemic inflammatory response and immune suppression exist simultaneously. Somatostatin has a dual immunomodulatory activity in these patients.

APACHE ; Case-Control Studies ; Female ; Humans ; Interleukin-10 ; blood ; Male ; Prognosis ; Prospective Studies ; Sepsis ; drug therapy ; etiology ; immunology ; Somatostatin ; therapeutic use ; T-Lymphocyte Subsets ; immunology ; Tumor Necrosis Factor-alpha ; blood

Objective To construct a recombinant plasmid expressing human microdystrophin gene fused with VP22 ofhmnan herpes simplex virus 1(HSV-1),and investigate the expression of microdystrophin in vitro and the characteristics of VP22-mediated protein transduction.Methods Full length HSV-1 VP22 gene was amplified by PCR from the plasmid pSINrep5-VP22 and cloned into the eukatyotic expression vector pAVXl to conslruct recombinant plasmid pAVP22.Microdystrophin cDNA was obtained from the recombinant plasmid pBSK-MICRO digested with Not Ⅰ,and the product Was inserted into plasmid pAVP22 to construct the plasmid pAVP22-MICDYS. 3T3 cells were transfected with pAVP22-MICDYS,and the expression of microdystrophin was detected by RT-PCR,Western blotting and immtmocytochemislry.The supematant of 3T3 cells transfected with pAVP22-MICDYS were collected to infect human mesenchymal cells(MSCs),and the expression of the fusion protein VP22-microdystrophin in the cells was detected using immunohistochemistry to assess VP22-mediated protein transduction. Results The recombinant plasmid expressing microdystrophin-VP22 fusion gene capable of in vitro expression of the fusion protein was constructed successfully.VP22 was shown to enhance the expression efficiency of microdystrophin in 3T3 cells and transduce VP22-microdystrophin fusion protein from 3T3 cells to human MSCs. Conclusion The recombinant plasmid expressing microdystrophin-VP22 fusion gene with protein transduction capacity provides an important basis for further study of the fusion protein in treatment of Duchenne muscular dystrophy.

OBJECTIVETo select the optimal pregnancy time window of embryonic pig skin precursor tissue for xenotransplantation and study its ability in wound repair.

METHODSSkin precursor tissues were obtained from pig fetus of fetal age of 35, 42, 56, 70 days, and were minced into microskin and transplanted to dorsal wounds of BALB/c nude mice, then they were covered with residual skin after plastic surgery of patients or adult pig skin (white). The characteristics of growth and development were observed after transplantation. Pathological examination was performed on 6 and 12 post operation weeks respectively to observe the tissue structure and tumorigenicity.

RESULTSSkin precursor tissues from fetal pig survived and developed after transplantation, and the microskin fused. New tissue area from skin precursor tissues with fetal age of 42 days was (47 +/- 6) mm2, which was higher than that of 35 days (18 +/- 8 mm2), 56 days (31 +/- 12 mm2), 70 days (20 +/- 8 mm2, P < 0.05). The skin precursor developed into "intact skin" with hair, sebaceous glands and sweat glands, and melanocytes were also detected in epidermis. The newly-grown skin tissue included epidermal and dermal layer, and obvious dermal papillae. Teratoma was not found after transplantation in skin precursor tissue with fetal age of 56, 70 days.

CONCLUSIONFetal pig skin precursor tissue with fetal age of 56 days can be used to repair wound as xenotransplantation.

Animals ; Fetal Tissue Transplantation ; Fetus ; Gestational Age ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Skin Transplantation ; Swine ; Transplantation, Heterologous ; Wound Healing