Objective To investigate the difference in ocular surface microbiota between dry eye patients and healthy subjects,and discuss the role of microbiota in dry eye.Methods Twenty cases of dry eye patients and 90 cases of healthy subjects were collected in the PLA General Hospital and Zhongshan Ophthalmic Center.The samples of conjunctiva impression cytology were collected from all subjects,and then metagenomic shotgun sequencing was performed following the DNA extraction.The differences in alpha diversity and metabolic pathways of the ocular surface microbiota between dry eye patients and healthy subjects were evaluated.Results There was no significant difference in alpha diversity of the microbial community between dry eye patients and healthy subjects (P =0.13).However,an increase of 15 species and a decrease of 10 species were detected on the ocular surface of dry eye patients.The enriched antibiotic resistance genes in dry eye patients were more than healthy subjects.Conclusion Although the alpha diversity of the microbial community on ocular surface between dry eye patients and healthy subjects are not distinguishable,a significant difference could be found in relative abundance and metabolic pathways,suggest that these specific microbiome may be related to the pathogenesis and disease progression of dry eye.

Animals ; HSP70 Heat-Shock Proteins ; metabolism ; Male ; Physical Conditioning, Animal ; Physical Endurance ; physiology ; Quadriceps Muscle ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism

Objective To detect the expression of ARK5 in hepatocellular carcinoma (HCC)tissue and hepatoma SMMC-7721 cells,and to investigate its effect on the growth of hepatoma cells.Methods The expression levels of ARK5 mRNA and protein were determined by RT-PCR and Western blotting in 30 cases of HCC tissue, paracarcinoma tissue,SMMC-7721 cells,and hepatic cells LO2.The SiRNA of ARK5 and negative control (NC) siRNA were constructed and transfected into the SMMC-7721 cells,and used as experimental group and negative control group;at the same time blank control group was set up. The proliferation activity and apoptotic rate of transfected cells were detected by MTT assay and flow cytometry (FCM).Results The PCR and Western blotting results showed that the expression levels of ARK5 mRNA and protein in HCC tissue and SMMC-7721 cells were significantly higher than those in paracarcinoma tissue and LO2 cells (P<0.05 ). The MTT assay results demonstrated that the inhibitory rates of growth of transfected cells in experimental group at 24,48 and 72 h were (19.39±5.42)%, (23.19±0.53)%,and (20.74±1.23)%;there were significant differences compared with blank control group and negative control group (P<0.01).The FCM results indicated that the apoptotic rate of the transfected cells in experimental group was (15.017±0.945)%,there were significant differences compared with blank control group (8.770%± 0.656 )% and negative control group (8.763%± 1.201%) (P<0.05 ). Conclusion The ARK5 expression level is significantly increased in HCC tissue and hepatoma SMMC-7721 cells;the inhibition of ARK5 expression could suppress the growth of hepatoma cells and induce apoptosis. So ARK5 maybe act as a cancer-promoting gene and induce hepatocellular carcinogenesis.

OBJECTIVEThis study aims to evaluate the quality of life (QOL) of patients who underwent resection of oral cancer and reconstruction by free anierolateral thigh perforator flaps (ALTF).

METHODSA total of 32 patients with oral and maxillofacial malignancies who had undergone the resection of oral cancer and reconstruction by ALTF were retrospectively analyzed. At 12 months postoperatively, the QOL of these patients was assessed by using the 14-item oral health impact profile (OHIP-14) and the medical outcome study short form-36 (SF-36) questionnaires.

RESULTSA total of 32 questionnaires were collected. In SF-36, the highest scoring domains were bodily pain (78.58 ± 14.82), physical functioning (72.08 ± 27.86), and the role of physical (60.00 ± 42.63), whereas the lowest scoring domains were role-emotional (41.67 ± 39.62), followed by mental health (50.75 ± 13.07) and health transition (54.17 ± 21.75). In OHIP-14, the lowest scoring domains were social disability (34.50 ± 11.32) and handicap (36.04 ± 12.05), indicating the functional recovery was better; and the highest scoring domains were physical pain (73.50 ± 18.96) and psychological discomfort (60.17 ± 20.66), indicating the functional recovery was worse.

CONCLUSIONThe ALTF is an ideal selection for the reconstruction of oral defects after cancer resection. In using this flap, the basic social need of patients after surgery can be satisfied. Moreover, the appearance and the functions of chewing, deglutition, and speech can be restored in varying degrees. Thus, ALTF can improve the patients' QOL.

Emotions ; Free Tissue Flaps ; Humans ; Mastication ; Mouth Neoplasms ; physiopathology ; psychology ; surgery ; Pain ; Perforator Flap ; Postoperative Period ; Quality of Life ; Reconstructive Surgical Procedures ; Retrospective Studies ; Surveys and Questionnaires ; Thigh

BACKGROUND: Stem cell transplantation in repairing infarct myocardium and in improving cardiac function has been widely accepted. However, whether transplanted cells and host cells formed an effective electricity and mechanical couple, whether a relevant independent electrical system with contractile function formed or whether severe malignant ventricular arrhythmia formed, are still unclear.OBJECTIVE: To investigate electrophysiological abnormaltiy and left ventricular remodeling in rats with myocardial infarction following allogenic bone marrow mesenchymal stem cell (BMSC) transplantation.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Experimental Center, Guangxi Medical University from December 2005 to October 2008.MATERIALS: A total of 120 healthy Wistar rats were equally randomized into normal control, sham operation, saline control and cell transplantation groups. Healthy Wister rats aged 1 month were selected to harvest bone marrow.METHODS: At the third passage, rat BMSCs were collected and treated with 5-aza, and differentiated into cerdiomyocytes.BMSCs were labeled with DAPI at 2 hours before transplantation. In the saline control and cell transplantation groups, rat models of myocardial infarction were established by ligating the left anterior descending coronary artery. In the sham operation group, the coronary artery was not ligated, but only braid. At 7 days following ligation, BMSCs in the cell transplantation group at 2×10-1/L were infused into the edge and center of myocardial infarct region by multipoint injection. Rats in the other three groups were subjected to an equal volume of saline.MAIN OUTCOME MEASURES: Electrocardiogram and cardiac electrophysiology were performed. Ultrasonic cardiography was used to detect left ventricular function. Infarct size was determined. DAPl-labeled donor cell migration and distribution was observed with a fluorescence microscope.RESULTS: BMSCs could differentiate into cardiacmuscle cell-like cells which were capable of pulsing spontaneously, expressing cardiactoponin T and forming myofilament in vitro. Compared with the saline control group, PR interval, QRS duration and ventdcular effective refractory period shortened, ventricular fibrillation threshold increased at 4, 8 and 12 weeks (P < 0.05); left ventricular internal diameter at end-systole reduced, and left ventricular ejection fraction and shortening traction was significantly increased (P< 0.05). At 8 and 12 weeks, infarct size was significantly smaller (P < 0.05). At 4 weeks, DAPl-labeled BMSCs could be seen under the fluorescence microscope, and still could he detected at 12 weeks. However, the fluorescence became weak with prolonged time.CONCLUSION: BMSCs have the plasticity of differentiating into cardiac muscle cell-like cells, which can modulate theelectrophysiological abnormality and left ventricular remodeling following myocardial infarction.

BACKGROUND:Studies have shown that bisphosphonates inhibit osteoclast resorption, but whether cathepsin K, a key cytokine of bone resorption, plays an effect has rarely been reported.
OBJECTIVE:To study the effect of bisphosphonate on capthesin K and bone resorption function during osteoclast differentiation.
METHODS:Osteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groups:control group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture and gene expression of capthesin K was detected by immunofluorescence method at 72 hours of culture. Western blot assay was used to detect capthesin K protein expression at 72 hours of culture.
RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group (P<0.01). Immunofluorescence showed expression of capthesin K was higher in the control group than the alendronate group (P<0.01);furthermore, the protein expression of capthesin K was also lower in the alendronate group than the control group (P<0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting gene expression of capthesin K.

BACKGROUND:Tartrate-resistant acid phosphatase is a specific marker for osteoclast differentiation and bone resorption, which is a sign of osteoclast maturity.
OBJECTIVE:To study the effect of alendronate on tartrate-resistant acid phosphatase related to osteoclast differentiation and bone resorption.
METHODOsteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groupcontrol group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture. Gene expression of tartrate-resistant acid phosphatase was detected by immunofluorescence method. Western blot assay was used to detect protein expression of tartrate-resistant acid phosphatase.
RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group (P<0.01). Immunofluorescence showed expression of tartrate-resistant acid phosphatase was higher in the control group than the alendronate group (P<0.01);furthermore, the protein expression of tartrate-resistant acid phosphatase was also lower in the alendronate group than the control group (P<0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting protein expression of tartrate-resistant acid phosphatase.

Objective To develop an effective way to evaluate the accurate platelet count in a patient with anticoagulants-induced pseudothrombocytopenia (PTCP).Methods It was studied that various anticoagulants effect on the platelets count for an infrequent patient with anticoagulants-dependent PTCP. When vitamin B6,aminophylline,gentamicin and amikacin were separately added to four anticoagulated blood samples from anticoagulants-dependent patient within 15 min after blood withdrawal,platelets count and morphological changes of blood cells after 4 hours of incubation at room temperature were investigated. The best anti-aggregating agent and its optimal concentration among them were explored.Results The four anticoagulants all could not inhibit the aggregation of the patient's platelets.Only amikaein among the above anti-aggregating agents can prevent and dissociate the aggregation of platelets without apparent morphological changes of blood cells and the platelet counts was stable within 4 hours after blood drawn when amikacin was added either before or after blood sampling.With increasing the concentration of amikaein,the platelet counts increase and then tend to be stable.The optimal concentration of amikacin is 5 mg/ml blood.Conclusions The supplementation of amikaein either before or after blood sampling is a useful method for the diagnosis anticoagulants-dependent PTCP and for the eva/uation of platelet counts in infrequent patients with anticoagulants-dependent PTCP.

Objective To investigate the suppressive effects of collagen from Cyanea nozakii on adjuvant induced arthritis inrat.Methods Rats with adjuvant arthritis received different do- ses of Cyanea nozakii collagen by intragastric administration for two weeks.Incidence and severity of arthritis were assessed by calculation of mean arthritis index,the concentrations of NO,MDA and activity of SOD in the serum were examined.Results Cyanea nozakii colla- gen at different doses could ameliorate the adjuvant induced arthritis,suppress the concen- trations of NO,MDA and increase the activity of SOD in the serum.Conclusion Cyanea noza- kii collagen has therapeutic efficacy in treatment of adjuvant arthritis rats,and the mecha- nism of Cyanea nozakii collagen is probably related to the antioxidation.

Two endophytic-bacteria isolates of G18 and F19 were isolated from the stem of Taxus chinensis. The G18 and F19 were respectively classified into Psudomonas sp. and Stenotrophomonas sp. based on biological characteristics and 16S rDNA sequence analysis. The bioactivity analysis showed that the fermented broths of the G18 and F19 exhibited antagonistic activities against three pathogenic bacteria, and had good antagonistic effectiveness to Verticillium dahliae and Colletotrichum gloeosporioides, respectively. The G18 can degrade salicylic acid, and the F19 can do dichlorvos.