BACKGROUND:Sensorineural hearing loss is mainly caused by missing or damaged hair cells in the inner ear. Application of adipose-derived mesenchymal stem cells to regenerate inner ear hair cells is an effective treatment for hearing loss. OBJECTIVE:To explore the feasibility of in vitro inducing adipose-derived mesenchymal stem cells to differentiate into inner ear hair cel-like cells in guinea pigs. METHODS:Adipose-derived mesenchymal stem cells from guinea pigs were isolated and cultured to the 3rd generation. cellphenotype was detected using flow cytometry. Cytokines were added for induction and differentiation by stages, including epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor-1, al-trans retinoic acid, brain-derived neurotrophic factor, neurotrophin 3. RESULTS AND CONCLUSION:Adipose-derived mesenchymal stem cells of guinea pigs cultured in vitro were fusiform and showed a swirled adherent growth. Passage 3 cells were positive for CDCD29 and CD44, but negative for CD34 and CD45. After induction, the cells were positive for nestin and GFAP positive at early stage;after 10-day continuous induction, the cells expressed Myosin VIIa and Math1, specific markers of hair cells, indicating that cytokines can directly induce adipose-derived mesenchymal stem cells to differentiating into inner ear hair cells in guinea pigs.

Objective To investigate radiation dose diffent type of anesthesia affected radiation dose when we do interventional therapy for child patent ductus arteriosus (PDA).Methods From 201 1 to 2014,we collected 50 children(3-6 years old)with PDA which received interventional therapy were included in the study.They were divided into two groups:group A (25 cases,male/fe-male=7/18,mean weight=1 5.32 kg±2.41 5 kg)underwent interventional therapy of PDA under general anesthesia,and group B (25 cases,male/female=13/12,mean weight=1 6.40 kg±2.056 kg)using local anesthesia.The surgery were operated by the same doctor,we used DSA children cardiovascular film AE mode (ped CARD)to monitor the surgery,image frames 1 5-30 f/s,used the non-ionic contrast agent (Iodixanol 320 mg I/mL)and recorded the child cumulative incidence of skin surface dose (AK),dose area product(DAP),and time of fluoroscopy,and do statistical analysis.Results All 50 cases were performed the surgery successfully. There were no significant difference of age,weight and gender between groups (age:t=1.924,P =0.06;weight:t =1.703,P =0.095;gender:χ2 =3.00,P =0.083).The cumulative incidence of skin surface dose (AK),dose area product (DAP)and time of fluoroscopy were (0.061±0.025)Gy,(5.08±2.19)Gy·cm2 and (3.15±1.16)min in A group,and (0.094±0.046)Gy,(8.41±3.587)Gy·cm2 , (6.86±3.27)min in B group.The sequence and image number of two groups were same.There were significantly differences of cumulative incidence of skin surface dose (AK),dose area product (DAP)and time of fluoroscopy between two groups (AK:t =3.152,P =0.003;DAP:t =3.957,P =0.000;time of fluoroscopy:t =5.346,P =0.000).The radiation doses of A group significantly lower than B group,compared with B group,the radiation dose were 40 percent [(1-5.08/8.41)%]lower in A group,the 1.1 7 times discrepancy of time of fluoroscopy between two group [(3.1 5-6.86)/3.1 5].Conclusion Radiation dose is associated with type of anesthesia. Compared with local anesthesia,radiation dose reduced 40% using general anesthesia during interventional therapy for PDA.

Objective To investigate cognitive impairment mechanism by studying dorsolateral prefrontal cortex connectivity in patients with primary insomnia.Methods Forty patients with primary insomnia and 50 healthy subjects from the Department of Neurology,People's Hospital of Zhengzhou University during the period April 2011 through April 2013 were included.The World Health OrganizationUniversity of California Los Angeles Auditory Verbal Learning Test (WHO-UCLA AVLT) and the digital pin test were applied to evaluate the subjects' word study ability and vigilance.Resting state functional magnetic resonance imaging was used to observe the connectivity of dorsolateral prefrontal cortex.Results The Pittsburgh Sleep Quality Index (2.00 (1.00,3.00)) and the Hamilton Anxiety Scale scores (13.00 (11.25,15.75)) of primary insomnia patients were significantly higher than that of healthy controls (11.00(9.00,13.00),1.00 (0,2.00),Z=-5.517,Z=-5.525,P＜0.01).Digital pin test efficiency (60.03％ ± 13.95％ vs 66.32％ ± 13.73％,t =2.142,P＜0.05) and WHO-UCLA word learning (10.11 ± 2.29 vs 11.95 ± 2.42,t =-3.493,P ＜ 0.01) of primary insomnia patients were significantly lower than that of healthy controls.Compared to the healthy controls,the right dorsolateral prefrontal cortex of primary insomnia patients exhibited decreased functional connectivity of the right prefrontal lobe (-2.610 3 ± 0.172 6,t =-3.504,P ＜ 0.05).The left dorsolateral prefrontal cortex of primary insomnia patients exhibited increased functional connectivity of the bilateral insular lobes and right prefrontal lobe (2.8204±0.326 5,2.371 7 ±0.106 6,2.492 6 ±0.052 8,t =4.032,t =3.340,t =3.037,P ＜0.05).Conclusions The ability of WHO-UCLA word study and the digital pin test efficiency have been shown to decline in patients with primary insomnia.The possible mechanism of cognitive impairment may be the abnormal dorsolateral prefrontal cortex connectivity in patients with primary insomnia.

Objective To investigate the value of dynamic contrast?enhanced MRI (DCE?MRI) in the differential diagnosis of glioblastoma and brain metastases. Methods Twenty patients with high grade gliomas and 20 cases patients with brain metastases proved by surgery and pathology were collected, and patients were examined with conventional MRI and DCE?MRI preoperatively. The ROIs were manually placed in solid parts of the tumors and their surrounding tissues to calculate Ktrans, Kep and Ve values. The Ktrans, Kep and Ve values differences for the solid part and surrounding tissues of the two brain tumors were compared by two independent sample t test. The correlation between Ktrans of the solid parts of the two brain tumors and Ktrans, Kep and Ve values of their surrounding tissues were studied by Pearson correlation analysis. Results The Ktrans, Kep and Ve values of glioblastoma were(0.258 ± 0.063)min-1,(0.398 ± 0.082)min-1, 0.632±0.084, the Ktrans, Kep and Ve values of brain metastases were(0.233±0.053)min-1,(0.357±0.042)min-1, 0.672±0.113. There were no significant differences between the glioblastoma and brain metastases for Ktrans, Kep and Ve values(t=-1.354,-1.982, 1.276, all P>0.05). The Ktrans, Kep and Ve values of surrounding tissues of glioblastoma were(0.093±0.032)min-1,(0.411±0.089)min-1, 0.107±0.021, the Ktrans, Kep and Ve values of surrounding tissues of brain metastases were(0.033±0.010)min-1,(0.204±0.045)min-1, 0.069±0.017. The Ktrans, Kep and Ve values of surrounding tissues between glioblastoma and brain metastases had significant difference (t=-7.978,-9.303,-6.203, all P<0.05). The Ktrans of glioblastoma were correlated with Ktrans, Kep and Ve values of their surrounding tissues (r=0.759, 0.464, 0.651, all P<0.05); The Ktrans values of brain metastases had no relationship with Ktrans, Kep and Ve values of their surrounding tissues (P>0.05). Conclusion The DCE?MRI can quantitatively display the microvascular permeability and accurately evaluate the damage of blood?brain barrier of glioblastoma and brain metastases, which has an important value in studying biological characteristics and differential diagnosis of the two brain tumors.

BACKGROUND:A large number of studies have confirmed that anterior approach and posterior approach for multilevel cervical spondylotic myelopathy were effective, but there is stil no conclusion in which one is better.
OBJECTIVE:To systematical y assess the clinical effectiveness and safety of anterior approach versus posterior approach for multilevel cervical spondylotic myelopathy.
METHODS:The databases such as The Cochrane Library (Issue 3, 2013), PubMed (from 1966 to March 2013), OVID (from 1950 to March 2013), EMbase (from 1966 to March 2013), Chinese Biomedical Literature Database (from 1978 to March 2013), WanFang Database (from 1998 to March 2013), China National Knowledge Infrastructure (from 1999 to March 2013) were electronical y searched and five relevant journals were searched by hand to col ect the randomized control ed trials or non-randomized control ed trials about the clinical effectiveness and safety of anterior approach versus posterior approach for multilevel cervical spondylotic myelopathy. Two reviewers independently screened the literature according to the inclusive and exclusive criteria, extracted the data, and assessed the methodological quality of included studies. Then the meta-analysis was performed by using RevMan5.2 software.
RESULTS AND CONCLUSION:A total of 11 control ed trials involving 814 patients were included. Meta-analysis results showed that, compared with posterior approach, postoperative Japanese Orthopaedic Association scores were better (P<0.000 01), improvement rate of neurological function was higher (P=0.000 3), the incidence of C5 root palsy was lower (P=0.007), but operation time was longer (P<0.000 01), amount of intraoperative bleedin g was larger (P=0.000 7), incidence of adjacent segments degeneration was higher (P=0.01), incidence of postoperative complications was higher (P<0.000 01) and the rate of secondary surgical procedures was higher (P=0.003) after anterior approach. Additional y, there were no differences between the two groups in the cervical range of motion (P=0.56). For quantity limitation and low methodological quality of included studies, this conclusion stil needs to be further proved by performing more high-quality and large-scale randomized control ed trials.

BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation, however there is rare report addressing the construction of the lentivirus targeted its genes to inhibit its expression in the spinal cord astrocytes.
OBJECTIVE:To construct recombinant lentivirus vector expressing smal interfering RNA against TERT gene and to evaluate its potential for inhibiting the TERT expression.
METHODS:After shRNA-TERT sequence was designed and synthesized, the sequence was amplified by PCR and then connected to plasmid pLentilox3.7U6-hTERT to construct recombinant plasmid. The recombinant plasmid was then transfected to DH5αcel s to screen positive colony, and the sequence was identified. The recombinant plasmid pLentilox3.7U6-TERT was transfected in 293T cel s, generating recombinant lentivirus Le-TERT. The titer of recombinant lentivirus was determined and Le-TERT was transfected into the rat spinal cord astrocytes. The expression of TERT in astrocytes was detected by RT-PCR, western blot and immunofluorescence assay.
RESULTS AND CONCLUSION:The gene sequencing analysis confirmed that, recombinant plasmid pLentilox3.7U6-TERT was successful y constructed. The real-time quantitative PCR, western blot analysis and immunofluorescence assay indicated that, after Le-TERT was transfected in the astrocytes for 4 days, the inhibition rate of TERT mRNA was (63.98±2.6)%, and Le-TERT was lowly expressed in the transfected astrocytes. Recombinant expression vector pLentilox3.7U6-TERT can produce the lentivirus at high titer and effectively inhibit TERT expression in the transfected astrocytes.

Objective To explore the effect of measure accuracy of crevasse diameter of atrial septal defect (ASD) by transthoracic echocardiography (TTE) on radiation dose during interventional therapy.Methods 79 cases ASD with interventional therapy in our hospital were analyzed retrospectively,according to the times which we chosen occluder device on the basis of defect diameter by TTE,we divided those cases into three groups:group A,the occluder device chosen well,and it took only one time to block success (50 cases);group B,it took two times to block success (22 cases);group C,because of the measurement error,it needed three times to block success (7 cases).Following data were separately recorded:①measurement values of TTE,size of the occluder device and times of occluder device exchange;② radiation dose [cumulative radiation dose (AK,Gy),area dose product (DAP,Gy · cm2),fluoroscopic time (T,min)];③patient gender,age and body mass.Results All the cases were blocked success,the coincidence rate of TTE and occluder device were 62.67 ％,28 ％,9.33 ％,but there were significantly differences of AK,DAP and T in three groups (FAK=12.119,P=0.000;FDAp=8.241,P=0.001;FT =12.777,P=0.000).It showed that the radiation dose and T of group C were the largest and the longest,and group A were the least and the shortest,while the radiation dose and T of group B ranged between A and C groups.There was no statistically difference between radiation dose for different gender,age,body mass(P≥0.050).Conclusion The times of block and radiation dose is most influenced by the coincidence rate of TTE and occluder device.Increase of the coincidence rate may decrease the times of block and reduce the radiation dose for surgeon and patient.

Objective To explore the changes of brain functional network in insomnia disorder (ID) during resting-state with voxel-based degree centrality (DC).Methods Forty-five subjects underwent resting-state fMRI scans,including 22 patients with ID (ID group) and 23 sex-,age-,and education-matched healthy volunteers (control group).The subjects' sleep quality and emotion state were assessed with Pittsburgh sleep quality index (PSQI),Hamilton anxiety scale and Hamilton depression scale.The resting-state fMRI data were analyzed with voxel-based DC.The intra-and inter-groups parameters were compared using t-test.Correlation analysis was performed between DC values of ID group and clinical parameters.Results Compared with control group,DC values increased in left parahippocampal gyrus,left hippocampus and bilateral precuneus (all P＜0.05),while decreased in left middle occipital lobe,left precuneus,left inferior frontal gyrus and left middle frontal gyrus in ID group (all P＜0.05).DC values of the left hippocampus in ID patients showed significantly negative correlation with the score of Pittsburgh sleep quality index (r=-0.46,P=0.047).Conclusion ID patients have abnormal DC distribution of brain functional network,therefore providing basis for understanding pathophysiological mechanisms of ID.

Objective To observe the alterations in brain gray matter volume(GMV)in idiopathic generalized epilepsy(IGE)patients with typical absence seizure. Methods Nine IGE patients with typical absence seizure and 16 healthy volunteers were prospectively recruited from People's Hospital of Zhengzhou University from September 2012 to January 2014. By using a 3. 0T magnetic resonance imaging(MRI)and voxel - based morphometry(VBM)method, their whole brain structures and their brain GMV were scanned and analyzed,respectively,then the changes in GMV were observed. Results Compared with healthy control group,brain GMV extensively decreased in IGE patients with typical absence seizure. Thirteen regions with significant differences were as follows:the right rectal gyrus(t = 3. 13,P ﹤0. 01),the left rectal gyrus(t = 4. 82,P ﹤ 0. 01),the right calcarine/ cuneus/ gyrus lingualis/ occipital gyrus/ inferior oc-cipital gyrus(t = 6. 86,P ﹤ 0. 01),right gyrus lingualis(t = 4. 01,P ﹤ 0. 01),the left gyrus lingualis/ inferior occipital gyrus(t = 3. 73,P ﹤ 0. 01),the left inferior occipital gyrus/ gyrus lingualis(t = 5. 42,P ﹤ 0. 01),the left middle occipi-tal gyrus(t = 3. 76,P ﹤ 0. 01),the right middle occipital gyrus/ superior occipital gyrus/ middle temporal gyrus( t =3. 85,P ﹤ 0. 01),left middle temporal gyrus/ superior temporal gyrus(t = 5. 06,P ﹤ 0. 01),the right precuneus/ cuneus (t = 3. 33,P ﹤ 0. 01),and the right superior parietal lo-bule(t = 3. 66,P ﹤ 0. 01),right precentral gyrus(t = 3. 44,P ﹤0. 01),right superior frontal gyrus/ paracentral lobule/ supplementary motor area(t = 3. 50,P ﹤ 0. 01). However,GMV increase was not found. Conclusions Brain GMV extensively decreased in 13 brain regions of IGE patients with typical absence seizure,and among them occipital lobe is the most significant.

Objective To analyze the complications of radical vaginal hysterectomy in patients with locally bulky cervical cancer.Methods All 258 patients of early cervical cancer (stage Ⅰ A2-Ⅱ A) selected in our hospital during the period of June 2011 to June 2016 undergoing radical vaginal hysterectomy and laparoscopic pelvic lymph node dissection were analyzed.All cases were divided into observation (locally bulky) and control (conventional) groups.Comparison analysis was performed for complications of the two groups and the influence of neoadjuvant chemotherapy on the complications of local bulky patients.Results The incidence of complications in the observation group was 26 cases (27.9％),higher than that in the control group of 28 cases (17.0％),the difference was statistically significant (P ＜ 0.05).The incidence of bladder injury (5 cases vs 2 cases) in observation group was significantly higher than that in control group (P ＜ 0.05).For patients with locally bulky early cervical cancer,the incidence rate of complications of the neoadjuvant chemotherapy group (15.8％) was significantly lower than the simple operation group (36.4％) (P ＜ 0.05).The incidence rate of ureteral injury in neoadjuvant chemotherapy group (0 cases vs 6 cases) was significantly lower than that in simple operation group (P ＜ 0.05).Conclusions Early cervical cancer patients with locally bulky (stage Ⅰ B2/ 1Ⅱ A2) had higher incidence of complications of radical vaginal hysterectomy,and neoadjuvant chemotherapy can reduce the incidence of surgical complications.