Objective To study the association between human papillomavirus (HPV) genotypes and viral load and clinical features of verruca vulgaris.Methods Tissue samples were collected from 48 outpatients with verruca vulgaris,and DNA was extracted from these tissue samples.To determine the genotype of HPV,PCR was performed to amplify the L1 fragment of HPV with universal primers followed by bidirectional sequencing and BLAST.The genotyping results were validated by PCR with type-specific primers.Real-time fluorescence-based quantitative PCR was conducted to measure the viral load of HPV,and hematoxylin and eosin (HE) staining to observe histological changes in these tissue specimens.Results The L1 fragment of HPV was amplified from 35 out of the 48 tissue specimens.Of the 35 L1-positive specimens,32 harbored HPV 7,1 harbored HPV 57,and 2 harbored both HPV 2 and HPV 7.Multiple lesions were observed on extremities in the patient infected with HPV 57,but on the head,face and trunk in the patients coinfected with HPV 2 and HPV 7.There were no significant differences in HPV viral load or vacuolated cell number between patients with single lesions and those with multiple lesions,or between patients with a clinical course of ＜ 6 months and those with a clinical course of 6-12 months.However,HPV viral load tended to decrease one year after the onset,and there was pronounced hyperkeratosis and less vacuolated cells in lesions of long duration (more than 2 years) compared with those of short duration (less than 2 years).Conclusions HPV 7 appears to be the most common HPV genotype associated with verruca vulgaris,and HPV 7 infection usually occurs on the head and face.For verruca vulgaris of less than 1 year,neither HPV viral load nor vacuolated cell number is associated with the count or clinical course of warts.

Objective To investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.

Objective To observe the expression of mitochondrial fission protein locus Fis1 and ultrastructural changes in the renal cells of rats with chronic fluorosis,and to reveal the mechanism in mitochondrial damage of the renal cells.Methods Sixty SD rats were randomly divided into 3 groups according to sex and body mass(20 in each group):control group,lower fluoride group and higher fluoride group.All the rats were fed with different doses of sodium fluoride in drinking water(0,10 and 50 mg/L,respectively).Six-month later,the expression of Fisl in renal cells was determined by real-time fluorenscence quantitative PCR and immunohistochemistry technology,the mitochondrial morphology of renal cells was observed under transmission electron microscopy (TEM).Results As compared with the control group(28.70 ± 12.41),Fis1 mRNA levels(91.48 + 34.83 and 582.09 ± 184.69) in renal cells of the lower fluoride and the higher fluoride groups were increased(all P ＜ 0.05).As compared with the control group(10.49 ± 7.66),Fisl protein levels(16.33 ± 10.26 and 21.50 ± 5.24) in renal cells of the lower fluoride and the higher fluoride groups showed a trend of increasing,the higher fluoride group was higher than that of the control group(P ＜ 0.05).By TEM,mitochondrial crest in renal cells of the lower fluoride and the higher fluoride groups was vague or disappeared,mitochondrial division section appeared.Conclusions Fluoride is a kind of toxicant that can cause damage to mitochondrion of renal cells,induce the expression of Fis1 in transcriptional and protein level,and lead to the obstacles of mitochondrial fusion-fission and ultrastructural abnormality of mitochondrion,which may play an important role in mechanism of mitochondrial damage in the renal cells of rats with chronic fluorosis.

AIM: To discuss the clinical applications of methods to localize nasal cut ends and the effects of Z-plasty in the surgeries for lower eyelid longitudinal laceration combined with lower lacrimal canaliculi disruption.
METHODS: From September, 2010 to October, 2013, a total of 37 patients ( 37 eyes ) with lower eyelid longitudinal laceration combined with lower lacrimal canaliculi disruption were operated for anastomosis of lacrimal canaliculi disruption and suture of lower eyelid longitudinal. Different methods to search for the nasal cut ends of lacerated lacrimal canaliculi, such as “under a microscope directly”, “guided by probing needle” and“pigtail curved probe”. Then, to repair lower eyelid longitudinal laceration with Z-plasty transposition flaps. Follow up was 3mo~2a after operation.
RESULTS: All nasal cut ends could be found successfully on 37 patients;Lacrimal duct unobstructed in 31 patients (83. 8%), improved in 5 patients (13. 5%), invalid in 1 patient (2. 7%),the overall successful rate was 97. 3%; the eyelids repair was satisfactory, small scars, the appearance and function was normal.
CONCLUSION: The nasal cut ends can be found successfully by “directly under a microscope”, “guided by probing needle” and“pigtail curved probe”;the effect of silicone drainage tube used as lacrimal canaliculi bracket is satisfactory; most patients gained excellent recovery for both appearance and function after Z-plasty.

OBJECTIVE: To investigate the resistance phenotype of acinetobacter baumannii and the expression of ade efflux pump gene. METHODS: Non-repetitive 51 strains of Acinetobacter baumannii were collected in Peking Union Medical Hospital between Feb. 2004 and Feb. 2005. The active efflux system adeB structural gene and sequence were identified by PCR. RESULTS: The tested strains were not susceptible to common broad-spectrum antibiotics. Multidrug resistant Acinetobacter baumannii carried adeB active efflux pump gene. CONCLUSION: The multiple-drug resistance of Acinetobacter baumannii is independent of ?-lactamase. It maybe related to other drug resistance mechanism. The effect of active efflux system is possibly one of the major mechanisms of multidrug resistance of acinetobacter baumannii.

Adolescent ; Altitude ; Ghrelin ; blood ; Humans ; Overweight ; blood ; Sleep

Objective To observe the effect of sustained release type Ⅰ collagen-vascular endothelial growth factor (VEGF) on healing of bone-tendon junction injuries.Methods Partial patellectomy was conducted in 72 rabbits divided equally into control group,type Ⅰ collagen group,and collagen type Ⅰ-VEGF group.The scaffold was planted into the bone-tendon interface.Animals were sacrificed at 4,8 and 12 weeks.New bone formation into the patella-patella tendon surface was detected using X-ray films and histological observations.Quality of bone healing was assayed using biomechanical testing.Results At postoperative 4,8 and 12 weeks,X-ray films showed bone formation of type Ⅰ collagen group [(4.1 ± 0.4) mm2,(12.1 ± 0.5) mm2,(13.0 ± 1.2) mm2 respectively] and of collagen type Ⅰ-VEGF group [(3.8 ± 0.4) mm2,(11.0 ± 0.5) mm2,(13.1 ± 1.0) mm2 respectively] were more than that of control group [(2.1 ± 0.6) mm2,(4.1 ± 0.3) mm2,(6.6 ± 0.6) mm2 respectively] (P ＜ 0.05).Histology identified few new bone,massive fibrocyte accumulation and disrupted alignment of tendon fiber in control group,massive new bone formation,neat and orderly alignment of collagen fiber tissues and massive aggrecan expression at postoperative 4 and 8 weeks (fibrous cartage repair in largely) in collagen type Ⅰ-VEGF group,and massive new bone formation but worse alignment of tendon collagen fibers and less aggrecan expression (fibrous repair in largely) in type Ⅰ collagen group.Biomechanical test showed the ultimate tensile strength increased over time in all groups,with significantly higher value at 12 weeks than that at 4 and 8 weeks.At the same time point,ultimate tensile strength ranged in an order as follows:collagen type Ⅰ-VEGF group ＞ collagen type Ⅰ group ＞ control group (P ＜ 0.05).Conclusion Sustained release type Ⅰ collagen-VEGF can accelerate early healing of bone-tendon junction injury and improve the histological and mechanical properties.

Objective To evaluate the relationship between sternum protection and bone marrow suppression in postoperative radiotherapy for esophageal carcinoma. Methods Total of 98 postoperative patients with thoracic esophageal carcinoma were randomly divided into experimental group (52 cases) and control group (46 cases). All patients were given intensity-modulated radiotherapy (IMRT), with the dose of 50-50.4 Gy. The patients in experimental group were irradiated by 6 fields (4-fields in front, 2-fields behind) which were crossed to avoid direct exposure to the sternum. The patients in control group were irradiated by 5 fields (3-fields in front, 2-fields behind) with front-middle of the field passing through the sternum. Concurrently all patients received 2 cycles of cisplatin chemotherapy. Results Dmean, V20 and V30 of the sternum in the experimental group were (20.21 ±3.60) Gy, (40.78 ±7.19) % and (33.78 ±9.44) %, which were lower than those in the control group [(30.91±5.21) Gy, (81.01±4.81) %, (51.60±6.84) %], respectively (P<0.05). However, the volume and dose distribution of lung, spinal cord and heart were similar between the two groups (P> 0.05). Both the incidence rates of bone marrow suppression at 14th day and 35th day after radiotherapy were significantly higher in the control group (52.2%, 73.9%) than those in the experimental group (28.8 %, 50.0 %) (P< 0.05), and the incidence rate of bone marrow suppression at 7th day after radiotherapy was similar between the two groups. Conclusion Protecting and sketching for sternum in postoperative radiotherapy for esophageal carcinoma can reduce the incidence of bone marrow suppression effectively, which would not increase the radiation dose in the lung, heart and spinal cord.

Objective To investigate the relationship between serum follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and clinicopathological features and prognosis of serous ovarian cancer retrospectively. Methods A total of 73 patients with serous ovarian cancer treated in the Department of Obstetrics and Gynecology of Tianjin Medical University General Hospital from January 2000 to December 2015 were included in this study. The relationship between serum FSH, LH, PRL and clinicopathological features was analyzed by Mann-Whitney U method. Kaplan-Meier (K-M) method was used to analyze survival rates of patients with different clinical features. Multivariate Cox proportional hazards regression analysis was used to analyze prognostic factors of serous ovarian cancer patients. Results The mean concentrations of serum FSH and LH were significantly higher in the>50 year-old group than those in the<50 year-old group (P<0.05). The mean concentrations of FSH and LH were significantly higher in menopause group than those in non-menopause group (P<0.05). There were no significant differences in serum levels of FSH and LH in patients with other different clinicopathological features (P>0.05). There was no significant correlation between serum PRL concentration and clinicopathological features (P>0.05). Analysis results showed that poor prognosis of patients was related with high serum levels of FSH (>40.13 IU/L), PRL (>14.96 μg/L) and FIGO stage (Ⅲ+Ⅳ) (P<0.05). There was no significant correlation between serum LH concentration and prognosis (P>0.05). COX regression analysis showed that the serum PRL>14.96 μg/L was risk factor for prognosis of serous ovarian cancer [HR(95%CI): 3.530(1.180-10.557),P=0.024]. Conclusion The serum levels of FSH and LH are significantly increased in postmenopausal women than those in menopause women. The serum level of PRL is correlated with the prognosis of serous ovarian cancer.

Objective To explore the effects of under-expression of large tumor suppressor 1 (LATS1) on activation of Hippo signaling pathway and differentiation, proliferation, migration of bone marrow mesenchymal stem cells (mMSCs) of micein vitro.Methods mMSCs of C57BL/6 mice were divided into normal control (MSC) group, empty vector control (MSC-GFP) group, LATS1-over-expressing (MSC-LATS1) group, empty vector without LATS1 shRNA control (MSC-shControl) group and LATS1-under-expressing (MSC-shLATS1) group. Lentiviral vectors with activated,inactivated LATS1 (the key molecule of Hippo signaling pathway) modifications and empty vectors were constructed and were used to infect mMSCsin vitro. The transduction efficiencies mediated by the lentiviral vectors were evaluated by fluorescence microscopy and flow cytometry. The mRNA expression of LATS1 was quantified by quantitative real-time polymerase chain reaction (qRT-PCR), and the protein expressions of LATS1, YAP (p-YAP), 14-3-3 were quantified by Western Blot to evaluate the activation of Hippo signaling pathway. Osteogenic and adipogenic differentiation of mMSCs were evaluated through measurement of Runx2, OSX and C/EBPα, PPAR-γ mRNA by qRT-PCR, as well as Alizarin Red S and Oil red O staining. Proliferation of mMSCs was evaluated using methy thiazdyl tetrazolium (MTT) assay. The scratch test and Transwell chamber test were used to analyze the horizontal and vertical migration ability of mMSCs.Results The transduction efficiencies mediated by the lentiviral vectors were 94.74%-96.10%. Compared with MSC-GFP group, the activation of Hippo signaling pathway was promoted in MSC-LATS1 group [LATS1 mRNA (2-ΔΔCT): 4.37±0.21 vs. 1.20±0.04, LATS1 protein (gray value): 2.21±0.06 vs. 1.09±0.10, p-YAP/YAP protein (gray value): 1.51±0.13 vs. 0.98±0.05, 14-3-3 protein (gray value): 1.92±0.18 vs. 1.10±0.09, allP < 0.05], osteogenic and adipogenic differentiation of mMSCs were decreased in MSC-LATS1 group [mineralization (A value):0.13±0.02 vs. 0.40±0.03, Runx2 mRNA (2-ΔΔCT): 0.51±0.02 vs. 0.98±0.09, OSX mRNA (2-ΔΔCT): 0.41±0.04 vs. 1.04±0.09, lipid accumulation (A value): 0.10±0.02 vs. 0.25±0.03, C/EBPα mRNA (2-ΔΔCT): 0.33±0.03 vs. 1.11±0.09, PPAR-γ mRNA (2-ΔΔCT): 0.29±0.02 vs. 1.04±0.10, allP < 0.05], the proliferation rate of mMSCs at 4-7 days was decreased in MSC-LATS1 group and so were the horizontal and vertical migration of mMSCs [wound healing rate: (18.65±3.53)% vs. (40.29±1.87)%, migrated cells (cells/MP): 35.99±6.18 vs. 103.67±17.77, bothP <0.05]. Compared with MSC-shControl group, the activation of Hippo signaling pathway was inhibited in MSC-shLATS1 group [LATS1 mRNA (2-ΔΔCT): 0.16±0.01 vs. 0.98±0.03, LATS1 protein (gray value): 0.38±0.03 vs. 1.04±0.07, p-YAP/YAP protein (gray value): 0.58±0.04 vs. 1.05±0.06, 14-3-3 protein (gray value): 0.14±0.02 vs. 1.02±0.09, allP < 0.05], osteogenic and adipogenic differentiation of mMSCs were increased in MSC-shLATS1 group [mineralization (A value): 0.93±0.13 vs. 0.44±0.05, Runx2 mRNA (2-ΔΔCT): 1.44±0.12 vs. 0.95±0.04, OSX mRNA (2-ΔΔCT):1.67±0.06 vs. 1.10±0.11, lipid accumulation (A value): 0.47±0.06 vs. 0.28±0.04, C/EBPα mRNA (2-ΔΔCT):3.98±0.61 vs. 0.99±0.10, PPAR-γ mRNA (2-ΔΔCT): 3.05±0.36 vs. 0.98±0.14, allP < 0.05], the proliferation rate of mMSCs at 3-7 days was increased in MSC-shLATS1 group and so were the horizontal and vertical migration of mMSCs [wound healing rate: (80.18±6.98)% vs. (46.18±1.01)%, migrated cells (cells/MP): 212.69±41.21 vs. 115.87±35.15, bothP < 0.05].Conclusions Under-expression of LATS1 promotes the differentiation, proliferation, migration of mMSCs by inhibition of Hippo signaling pathwayin vitro.