Objective To investigate the effects of dedicator of cytokinesis 1 (Dock180) knockout on proliferation and apoptosis in rat derived H9C2 cardiomyocytes and their mechanisms.Methods A single guide RNA (sgRNA) targeting rat Dock180 gene was designed and constructed using CRISPR/Cas9 system.A plasmid contained above sgRNA was packaged into lentivirus and selected to knockout Dock180 in the cardiomyocytes.A single clone of cardiomyocyte with Dock180 knockout was established.Cardiomyocytes were divided into negative lentivirus group (Cas9, A group), Dock180 knockout group (B group), Cas9 lentivirus hypoxia group (C group), Dock180 knockout hypoxia group (D group).The expression of Dock180 mRNA was examined by RT-PCR, and relevant proteins were detected by Western blot.The cell proliferation rate of the cardiomyocytes was determined by MTT, and the apoptotic rate was measured by flow cytometry.Results Dock180 mRNA and protein were absent in B andD groups.Compared with A and C groups, p-ERK1/2 and Bcl-2 protein expression and cell proliferation rate were lower in B and D groups respectively (P<0.01), while Bax protein expression and cell apoptosis rate were higher in B and D groups respectively (P<0.05, P<0.01);Compared with A group, Dock180 mRNA and protein, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were reduced, while Bax protein and cell apoptosis rate were increased in C group(P<0.05,P<0.01).Compared with B group, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were decreased, while Bax protein and cell apoptosis rate were increased in D group(P<0.05,P<0.01).Conclusions Dock180 knockout with CRISPR/Cas9 can inhibit proliferation and promote apoptosis via p-ERK1/2, Bcl-2 and Bax in H9C2 cardiomyocytes.

ObjectiveTo investigate the cause,diagnosis and treatment measures of neonatal pneumothorax.MethodsThe clinical data of 98 cases with neonatal pneumothorax in our hospital from Mar 2005 to Mar 2010 were reviewed retrospectively.ResultsThere were 27 premature neonates and 71 full-term neonates in the 98 cases.Ten cases were spontaneous pneumothorax and 88 cases were non spontaneous pneumothorax.After the treatnent of primary disease,pleural aspiration,chest drainage and mechanical ventilation,88 cases were cured,8 cases were improved and 2 cases died.ConclusionNeonatal pneumothorax is related with cesarean section,improper use of endotracheal intubation,meconium aspiration,premature hyaline membrane disease using pulmonary surfactant and ventilator therapy.Most pneumothorax is due to lung disease.The prevention of preterm birth and asphyxia can reduce the incidence of neonatal pneumothorax.

Objective To investigate the effect of platelet-rich plasma (PRP) on starting of wnt3 gene and klotho gene of adipose-derived stem cells (ADSCs) of rabbit.Methods Epididymal adipose tissue stem cells were obtained from New Zealand white rabbits,and the cells identified by morphology and inducing differentiation,and the cells were cultured to the fourth generation,PRP and PPP (platelet-poor plasma) were prepared by traditional centrifugal method from abdominal aortic of rabbit; ADSCs were cultured in culture medium containing PRP (experimental group),PPP (control group) and all medium (blank group) for each 5％ for 24h,48h and 72h.Cells of each group were dissociated and total RNA extracted.Effects of the starting of wnt3 gene and klotho gene were detected by RT-PCR.Results Primary ADSCs of rabbit grew in the way of long spindle swirly.The results of oil red O and alizarin red staining of the ADSCs were positive.Expression of wnt3 gene and klotho gene in the experimental group significantly increased from the results of RT-PCR (P＜0.05).Conclusions PRP can promote proliferation of the ADSCs of rabbit and increase the expression of wnt3 gene and klotho gene significantly.

Aim To investigate the effects of taxol on apoptosis in Hela cell and the mechanism of apoptosis. The apoptotic cells were detected by TUNEL, HE, eletronic micrpscopy and flow cytometry method. The expressions and activity of apoptosis associated proteins such as PCNA and caspase-3 were examined using S-P and enzyme histochemistric method.The results followed as: HeLa cells exposed to Taxol undergo cell death, presenting morphological and biochemical characteristics typical of apoptosis and the apoptotic cells increased with time and concentration. In contrast to untreated Hela cells, which express low PCNA, Ones treated with Taxol expressed high amounts of PCNA. Conclusion Taxol may induced apoptosis in Hela cell. The apoptosis induced by taxol is related to the increase of PCNA protein and activity of caspase-3.

Objective To evaluate the effect of ion chamber sensitive volume on absolute dose verification in CyberKnife plan.Methods Solid water phantoms were scanned by a CT scanner, single-field plan, multi-field isocentric plan and sequential optimized plan were designed by the treatment planning system.Absolute doses were measured at the specified point in each plan using the ion chambers with sensitive volumes of 0.007 cm3(A16), 0.24 cm3(A12 s), and 0.6 cm3(PTW30013) and compared with calculated values.Results For the single-field plan, the relative error increased as the aperture size of collimator decreased;with relative errors within ±2%, the smallest aperture sizes of collimator were 12.5 mm (A16), 25 mm (A12 s), and 30 mm (PTW30013).For the multi-filed isocentric plan, the relative errors were 0.26%±3.90%(A16),-6.28%±14.33%(A12 s), and-9.41%±14.10%(PTW30013).For the sequential plan optimized with 15 mm cone, the relative error was 0.79%±1.43%;for the sequential plan optimized with 7.5 mm cone, the relative error was 2.01%±8.39%.In absolute dose verification for clinical plans, there was no significant difference between the results measured by these ion chambers (P=0.985).Conclusions There is no significant effect of ion chamber sensitive volume on absolute dose verification in CyberKnife plan under the following two situations:(1) the collimator with a relatively large aperture is used;(2) the sensitive volume of ion chamber is totally covered by the prescription isodose line.

OBJECTIVE:To compare the efficacy,safety,vascular endothelial growth factor(VEGF)and matrix metallopro-teinase-2 (MMP-2) of docetaxel combined with carboplatin and paclitaxel combined with cisplatin (DDP) in the treatment of ad-vanced ovarian cancer. METHODS:120 patients with advanced ovarian cancer were randomly divided into docetaxel combined with carboplatin group(60 cases)and paclitaxel combined with DDP group(60 cases). Docetaxel combined with carboplatin group received 70 mg/m2 Docetaxel injection,intravenous infusion of 1 h,d1;50 mg/m2 carboplatin injection,intravenous infusion of 1 h,d2. Paclitaxel combined with DDP group received 135 mg/m2 Paclitaxel injection,intravenous infusion of 24 h,d1;30 mg/m2 DDP for injection,intravenous infusion,d3;60 mg/m2 Paclitaxel injection (a maximum of 2.0 m2) by intraperitoneal infusion,d8. 3-week was regarded as 1 treatment course,and it lasted 6 courses. Clinical efficacy,VEGF,MMP-2,progression-free survival, overall survival before and after treatment,mortality rate within 2 years of treatment and the incidence of adverse reactions in 2 groups were compared. RESULTS:There were no significant differences in the objective response rate,disease control rate,mortal-ity rate,incidence of adverse reactions between 2 groups(P>0.05). The progression-free survival in docetaxel combined with car-boplatin group was significantly longer than paclitaxel combined with DDP group,the difference was statistically significant (P<0.05). Before treatment,there were no significant differences in VEGF and MMP-2 level between 2 groups(P>0.05). After treat-ment,VEGF and MMP-2 level in 2 groups were significantly lower than before,and VEGF at different time points and MMP-2 level after 4 weeks,8 weeks and 12 weeks of treatment in docetaxel combined with carboplatin group were lower than paclitaxel combined with DDP group,the differences were statistically significant(P<0.05). CONCLUSIONS:Docetaxel combined with car-boplatin and paclitaxel combined with DDP shows similar efficacy and safety in the treatment of advanced ovarian cancer,but docetaxel carboplatin combined with is superior to paclitaxel combined with DDP in reducing VEGF and MMP-2 and improving pro-gression-free survival.

Objective To assess the value of preoperative neutrophil-lymphocyte ratio ( NLR) for predict the prognosis in patients with high grade T1 bladder.Methods From January 2004 to December 2014, the data of 307 patients diagnosed as bladder cancer of Stage 1 and high grade after undergoing TURBT were analyzed, including gender, age, smoking status, tumor number and size, hydronephrosis, intravesical instillations and preoperative blood transfusion of 307 patients diagnosed as bladder cancer of stage 1 and high grade after undergoing TURBT were analyzed retrospectively.All patients were primary urothelial carcinoma.According to preoperative NLR,patients were divided into the low NLR group( NLR≤2.42,n=197) and the high NLR group(NLR >2.42,n =110).Recurrence-free survival (RFS) and progression-free survival ( PFS) were calculated according to the Kaplan-Meier model and compared by the log-rank model.Cox regression models were used for multivariate analyses of the association between NLR and bladder cancer, then the prognostic factors affecting RFS and PFS were evaluated.Result of these 307 patients, the low NLR group accounted for 64.2%(197/307), and the high NLR group accounted for 35. 8%(110/307).The mean follow-up period was 71(range, 1-123)months.The recurrence rate in the low NLR group and the high NLR group recurrence rate were 19.2%( 38/197 ) and 34.5%( 38/110 ) respectively, RFS were 73.0(range, 2-123)months and 67.5(range, 1-122)months respectively.The progression rates were 4.1%(8/197) and 10.9%(12/110) respectively.The recurrence and progression rates in the high NLR group is higher than those in the low NLR group(P<0.01 and P=0.008), and RFS was shorter( P=0.002).Multivariable Cox regression analysis showed that NLR>2.42(P=0.007,HR=1.912)and hydronephrosis (P<0.01, HR =2.485 ) are associated with higher risk of recurrence.Conclusion Elevated preoperative NLR is an independent predictor of RFS and PFS in patients with high grade T1 bladder cancer.

OBJECTIVETo investigate the facial aging analysis of 1200 Han adult people in Dalian with iris diameter, so as to provide reference for facial rejuvenation in Han people.

METHODS1200 facial digital photographs from the second-generation ID card of China were stratified randomly selected as the research sample, excluding facial deformity, tattooing eyebrow, tattooing lid margin, ptosis, lip grain and eyebrow deformity cases, as well as the cases whose join point of upper eyelid and lower eyelid not exposed at the nasal side. These photos were divided into six cohorts: female aged 18, 38, and 68 years old; and male aged 18, 38, and 68 years old, with 200 cases in each cohort. The individual iris diameter was divided into 20 equal parts in each photograph, with each part marked one unit as individual iris diameter ruler. Ten values were made from a horizontal plane between the medial canthus to ten points at one face ( the line between two medial canthus is used as the horizontal line, circumocular and facial soft tissue were measured with individual iris diameter ruler on the photographs), then the results were analyzed with Adobe Photoshop software.

RESULTSThere are statistically significant differences in eyebrow height (36.42 ± 4.22 unit in 18 years old male group, 40.22 ± 6.90 unit in 38 years old male group, 34.83 ± 9.39 unit in 68 years old male group; 37.59 ± 6.72 unit in 18 years old female group, 41.09 ± 5.15 unit in 38 years old female group, 36.84 ± 9.45 unit in 68 years old female group), palpebral fissure height, physiognomic external canthus height, palpebral fissure width, pupil height and other items (P < 0.05).

CONCLUSIONS(1) The brow position rises to the peak level at middle age (38 years old group), then drops down gradually with aging. (2) The physiognomic external canthus moves towards the nasal side and caudal side with aging. (3) Eyeball moves towards caudal side with aging. (4) The soft tissue around bilateral angle of mouth, nasal tip and submaxilla moves towards caudal side with aging. (5) The measurement of individual iris diameter ruler can apply to analyze the aging changes of facial soft tissue, and is more suitable for the case when facial photographs are taken at different distances.

Adolescent ; Adult ; Age Factors ; Aged ; Aging ; China ; ethnology ; Cohort Studies ; Eye ; anatomy & histology ; Eyebrows ; anatomy & histology ; Face ; anatomy & histology ; Female ; Humans ; Iris ; anatomy & histology ; Lip ; anatomy & histology ; Male ; Nose ; anatomy & histology ; Photography ; Rejuvenation ; Sex Factors

Objective To investigate whether the effect of puerarin on right ventricle hypertrophy of pulmonary hypertensive rats induced by chronic hypoxia-hypercapnia was related to new peptide Apelin or its receptor(APJ).Methods Thirty male Sprague-Dawley rats were randomly divided into three groups, they are control group,hypoxia-hypercapnia 4-week model group,and hypoxia-hypercapnia 4-week plus puerarin group.The concentrations of Apelin-36 protein in plasma and homogenate of right ventricular muscle were detected by radioimmunoassay.The mRNA expressions of Apelin and APJ in right ventricu- lar muscle were measured by semi-quantitive reverse transcription polymerase chain reaction(RT-PCR). Results The weight ratio of right ventricle to left ventricle plus septum[RV/(LV+S)] in model group was significantly higher than that in control group(P0.05).The plasma concen- tration of Apelin-36 protein in model group was significantly higher than that in control group(P

OBJECTIVETo summary the application of the extended free lateral arm flaps for buccal soft tissue reconstruction after buccal cancer.

METHODSFrom January to August 2011, three patients underwent the operation of buccal defect reconstruction using the extended free lateral arm flap in one-stage. PRCA was identification with the Doppler probe. According to the mark of PRCA, size and shape of defects, the flaps were designed and extended to the lateral epicondyle of humerus. The flap size ranged from 9 cm x 5 cm to 10 cm x 6 cm with a pedicle of 10 cm in length. The wounds at donor sites were closed directly.

RESULTSVascular crisis happened in one case due to local negative pressure, which resolved after emergency management. All the flaps survived completely. The patients were followed up for 6 to 10 months with no recurrence. Both the esthetic and functional results were satisfactory. Two cases suffered from numb feeling in donor sites which alleviated six months later.

CONCLUSIONSThe extended free lateral arm flap has reliable blood supply with appropriate thickness. It is an optional method for reconstruction of buccal defects after ablation of buccal cancer.

Aged ; Arm ; surgery ; Cheek ; surgery ; Facial Neoplasms ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps