Objective:To investigate the effect of GRP94 gene expression down-regulation on the proliferation in human lung carcinoma A549 cells and possible mechanism.Methods:Expression of GRP94 was examined in 4 lung carcinoma cells and two lung bronchial epithelial cells by qRT-PCR and Western blot.Chemically synthesized siRNA targeting GRP94 gene and transfected into A549 cells used by Lipofectamine2000.The mRNA and protein expression levels of GRP94,c-myc and cyclinD1 were detected by qRT-PCR and Western blot.CCK8 assay and Colony formation assay were used to detect the effect of specific GRP94 siRNA on cell proliferation.Results:The levels of GRP94 expression in the four lung carcinoma cells were significantly higher than that in two lung bronchial epithelial cells (P<0.05).GRP94 siRNA significantly downregulated the levels of GRP94 mRNA and protein in A549 cells, and the downregulation evidently suppressed cell proliferation.However, The c-myc and cyclinD1 expression levels in A549 cells in siRNA-GRP94 group was significantly lower than that in siRNA-NC group (P<0.05).Conclusion:The levels of GRP94 expression in lung carcinoma cells were significantly higher than that in normal lung epithelial cells.Knockdown GRP94 expression significantly inhibit the level of lung carcinoma A549 cells proliferation,which may act via influencing the expression of c-myc,cyclinD1.

Chromatography, Gas ; methods ; Chromatography, High Pressure Liquid ; methods ; Humans ; Hypnotics and Sedatives ; blood ; Pentobarbital ; blood ; Pharmaceutical Preparations ; blood ; Sensitivity and Specificity

Objective To explore the therapeutic effect of infrabrow excision blepharoplasty combined with botulinum toxin A (BTXA) injection on periorbital rejuvenation.Methods A total of 146 females were involved in this study.The incision was designed,hiding at the lower edge of the brow.Eye orbicularis was folded,eyelid lift and some of the skin removed during operation,and simultaneously the shape of eyebrow was better adjusted.After suture cutting,BTXA was injected at the glabella and the lateral of canthus.Results All the 146 cases were followed up for 3 to 12 months.The blepharotosis symptom was improved effectively,the periocular wrinkles were alleviated obviously and no complications occurred in these patients.Conclusions Combining Infrabrow excision blepharoplasty with BTXA injection in periocular treatment is safe,convenient and effective.It is a good clinical approach to make periorbital rejuvenation.

Objective To review the experience in managing severe major jugular vascular injuries. Methods Fifteen cases (11 men and 4 women, aged 18～49 years) treated by operation were collected. The injuried vessels included common carotid artery, internal carotid artery, subclavian artery and accompanying veins, carotid reconstruction was performed by jugular vein transplantation and followed by wrapping with PTFE artificial blood vessel in 15 cases. Result All the fifteen patients were cured and the vasculars were well-functioning 3～12 weeks after the surgery. Conclusion The mortality of severe large cervical vascular injuries is very high because of acute bleeding, cerebral ischimia and suffocation. In managing this catastrophe, keeping the airway clear and stop bleeding effectively are fundamental. Good knowledge and expertise in vascular and microvascular surgery are mandatory for the success of the procedure.

Objective Curcumin and MS-275,an inhibitor of histone deacetylase (HDAC),are promising anti-tumor agents. The aim of present study was to investigate the mechanisms of apoptosis induced by combined use of MS-275 in low dosage and curcumin in prostate cancer cell line DU145. Methods MTT assay was used to evaluate the lethal effect on DU145 cells by solitary use of MS-275 and or combined with curcumin. The changes in cell life cycle was detected by flow cytometry. The expressions of the survival signaling pathways were determined by Western blotting. Results Solitary application of MS-275 or curcumin may inhibit the growth of DU145 cells in a time and dose-dependent manner. The combination of MS-275 (1?mol/L) and curcumin (20?mol/L) exhibited obvious cytotoxic effect on cell viability,which was only 45.9% 48h after the combined treatment. Cell cycle assay showed that the combination of MS-275 and curcumin resulted in obvious appearance of sub-G1 phase in DU145 cells,implying that the cell apoptosis had been induced. The results of Western blotting showed that after treatment of MS-275 or curcumin singly,the phosphorylation level of Akt and ERK kinase declined slightly,however,when MS-275 and curcumin were used together,there appeared a prominent inhibitory effect on Akt and ERK kinase,indicated by a sharp decline of their phosphorylation level,and at the same time,the level of cleaved PARP,a hallmark of apoptosis,was increased in DU145 cells. Conclusion Combined use of MS-275 and curcumin may exert a synergistic cytotoxic effect on the viability of DU145 cells,and exhibit an inhibitory activity on Akt and ERK kinases to induce apoptosis.

Objective To study the effects of ultrasound mediated microbubbles destruction on endothelial cells transfected with lipofectamine DNA complexes.Methods One hour after addition of pIRES2 EGFP hVEGF gene and lipofectamine,endothelial cells on 6 well plates were exposed to ultrasound for 30 s,60 s,or 90 s in the presence(10 ?l,100 ?l,500 ?l) or absence of ultrasound contrast agent.The transducer (Sonos 5500,S3, 1.3 MHz,MI 1.0 ) was submerged in water and held at a distance of 1 cm.Expression of the marker gene EGFP was observed by fluorescent microscopy 2 days later.Results Ultrasound treatment of endothelial cells on plates for 30 s and 60 s yielded significant increase in transfection rates without damaging the cells,but 90 s treatment killed most of the cells.Ultrasound contrast agent was able to significantly increase endothelial cells transfection rate by enhancing cavitation effects at a concentration of 100 ?l,and damaged most cells when applied at a concentration of 500 ?l.Conclusions Ultrasound mediated destruction of microbubbles enhances gene expression after lipofectamine mediated transfection of endothelial cells in vitro; and it may represent an improved avenue for therapeutic gene delivery in vivo.

Objective To research the removal of the low concentration of formaldehyde in the indoor air by using copper sulfate.Methods The low concentration of formaldehyde(10.0 mg/L)in the indoor air was determined by the way of MBTH spectrophotometry.The influence of pH,chelon and concentration on the removal of different concentration formaldehyde was investigated by the way of chemisorption.Results When pH was 11.99,12.86,13.08 and 13.42,using copper sulfate,the removal rate of 10.0 mg/L formaldehyde was 43.82%,62.75%,69.21% and 73.40% respectively.When the concentration of copper sulfate was at 3.0 g/L,5.0 g/L,7.0 g/L and 10.0 g/L,the removal rate was 51.43%,73.40%,66.36% and 62.18% respectively in the condition of pH=13.42.When used potassium sodium tartrate and EDTA as the ehelon,pH=13.42,concentration of copper sulfate was 5.0 g/L,the removal rate of 2.0 mg/L formaldehyde was 77.21% and 62.51% respectively,that of 10.0 mg/L formaldehyde was 86.54% and 73.40% respectively,that of 100.0mg/L formaldehyde was 96.71% and 91.32% respectively.Conclusion Using potassium sodium tartrate as the chelon,at pH=13.42,5.0 g/L copper sulfate can produce a good removal efficiency for indoor low level formaldehyde.

OBJECTIVETo investigate the effects of free and pedicled thoracodorsal artery perforator (TDAP) flaps for repairing skin and soft tissue defects in limbs, neck, axillary and shoulder.

METHODSFrom October 2009 to Auguest 2011, 16 TDAP flaps were used to repair skin and tissue defects. Among them, five ipsilateral pedicled flaps were used to repair wounds in neck, axillary and shoulder. 11 free TDAP flaps were used to repair the wounds with bone or tendon exposure. In 12 cases, the flaps were pedicled with thoracodorsal artery and vein-lateral branches-perforators, in 4 cases, pedicled with thoracodorsal artery and vein-serratus anterior muscular branches-perforators. The deep fascia, the latissimus dorsi and thoracodorsal nerve were not included in all flaps. The flaps size ranged from 10 cm x 5 cm to 26 cm x 10 cm.

RESULTSAll 16 flaps survived completely with primary healing both at donor site and recipent area. After a follow-up of 3 to 24 months, all flaps gained good texture and appearance. Only linear scar was left at donor area. The shoulder could move freely.

CONCLUSIONSTDAP flap has good texture, long vascular pedicle,and reliable blood supply, leaving less morbidity at donor site. The latissimus dorsi and thoracodorsal nerve are also preserved. The pedicled TDAP flap is an ideal flap for repairing the ipsilateral skin and soft tissue defects of the neck, shoulder, axillary. The free TDAP flap is suited for repairing skin and soft tissue defects of the extremities.

Arteries ; Axilla ; Humans ; Muscle, Skeletal ; Perforator Flap ; transplantation ; Surgical Flaps ; blood supply ; transplantation ; Thoracic Wall ; Wound Healing ; Wounds and Injuries ; surgery

OBJECTIVETo observe the effect of cold or hot properties of traditional Chinese medicines (TCM) on biological effect indexes, and analyze the contribution of variables on cold or hot properties, in order to preliminarily establish the discrimination mode for the biological effects of cold or hot properties.

METHODRats were randomly divided into the blank control group, cold TCM groups (Coptidis Rhizoma, Scutellariae Radix, Phellodendri Cortex, Gardeniae Fructus, Sophorae Flavescentis Radix and Gentianae Radix) and hot TCM groups (Aconiti Lateralis Preparata Radix, Zingiberis Rhizoma, Alpiniae Officinarum Rhizoma, Zanthoxyli Pericarpium, Cinnamomi Cortex and Evodiae Fructus), and orally administered with 10 mL x kg(-1) of corresponding TCM water decoctions for 30 d, twice a day. Altogether 53 biological effect indexes correlated to cold or hot properties of traditional Chinese medicines were founded by searching literatures. The data warehouse were established by using data-mining software Clementine12.0. Data of the blank control group, cold TCM groups (Coptidis Rhizoma, Phellodendri Cortex, Gardeniae Fructus, Sophorae Flavescentis Radix, Gentianae Radix) and hot TCM groups (Aconiti Lateralis Preparata Radix, Zingiberis Rhizoma, Alpiniae Officinarum Rhizoma, Zanthoxyli Pericarpium, Cinnamomi Cortex) were selected into a training set. C5.0 algorithm and C&R classification and regression algorithm were adopted to define the importance of variable, create the decision trees, and test hot or cold properties of Evodiae Fructus and Scutellariae Radix.

RESULTAccording to C&R classification and regression algorithm, SDH activity of livers was the most important hot or cold property, with the significance closed to 30%. It was followed by triglyceride, liver Na' -K' -ATPase enzyme, muscle glycogen and platelet distribution width, with the accuracy up to 97.39% in models. C5.0 algorithm showed that liver SDH activity was the most important hot or cold property, with the significance closed to 40%. It was followed by triglyceride, GOT, muscle glycogen and liver Na(+)-K(+)-ATPase enzyme, with the accuracy up to 98.26% in models. The possibilities that Evodiae Fructus is in hot property and Scutellariae Radix is in cold property were 100. 00% and 77.78% by using both C&R classification and regression algorithm and C5.0 algorithm.

CONCLUSIONThe SDH activity of liver is the most important biological effect index to distinguish cold and hot properties of TCMs. The discrimination pathway or mode of cold and hot properties is closely related to energy metabolism.

Algorithms ; Animals ; Drugs, Chinese Herbal ; classification ; pharmacology ; Fruit ; chemistry ; Liver ; drug effects ; metabolism ; Liver Glycogen ; metabolism ; Male ; Medicine, Chinese Traditional ; methods ; Outcome Assessment (Health Care) ; methods ; Phytotherapy ; classification ; methods ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; classification ; Random Allocation ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Succinate Dehydrogenase ; metabolism ; Triglycerides ; metabolism

Objective With the method of detecting hepatitis B virus deoxyribonucleic acid(HBV-DNA) gene in breast milk,in ordered to diagnose whether there is HBV in breast milk in those mothers whose HBsAg are positive.Methods Collected breast milk from 187 mothers whose HBsAg were positive,and complified HBV-N gene in breast milk by Nested-Polymerase chain reaction (Nested-PCR).Results Nested-PCR could detect HBV-DNA in breast milk,and the positive rate was 3.2%.Conclusion The method of detecting HBV-N gene in breast milk by Nested-PCR can detect HBV-DNA in breast milk,and it can be an laboratory evidence for whether breast feeding or not.