In older to study the interrelationship between laryngeal carcinoma and human papillomavirus (HPV). The distribution and expression of genome types of HPV in laryngeal carcinoma were investigated. HPV DNA of 160 cases of fresh tissue samples with different lesions of the larynx was detected by applying consensus primers and multiple primers of polymerase chain reaction (PCR) . The consensus primers used were able to detect 9 types of HPVs DNAs such as HPV6, 11, 16, 18, 31, 33, 34, 42 and 58. The results turned out to be as follows: The rate of positive cases with HPV infection was 49.3% (35/71) for the group of laryneal carcinoma, 22.7% (5/22) for the group of neck metastatic lymphnode, 11.8% (2/17) for precarcinomous lesions and 6.7% (2/30 ) for the group of vocal cord polypus, 20 case of normal laryngeal tissue adjacent to the carcinoma was HPV DNA negative. The results demonstrated that the occurrence and development of laryngeal carcinoma were closely related to HPV infection.

Objective To understand the types of drug use,medication habits,medical treatment behavior and drug management of urban families in Hefei,and to provide background information for drug safety education and scientific medical education.Methods The questionnaire design covers 32 issues,some teachers and parents of 2 training institutions,part of the staff of the 2 administrative institutions and the residents of the 3 residential areas were investigated.320 questionnaires were distributed,and valid questionnaires were collected from 306 copies.Results 96.1% of Hefei urban family had standing drugs,anti cold drugs was the highest proportion(87.3%),followed by antibacterial drugs(51.0%),the main channel was the purchase pharmacy(89.2%) and hospital(36.3%).When the family was ill,65.7% of the respondents first ate home medicine;the proportion of chosing a large hospital up to 83.3%.74.5% of respondents believed that antibiotics abuse.Up to 83.3% of respondents had been performed intravenous infusion therapy when illness,55.9% of the respondents thought intravenous infusion therapy most unnecessary,and only 5.9% of the respondents thought intravenous infusion therapy was dangerous.Consulting a doctor when they had a problem in 67.7%,followed by the internet query(44.1%) and to the pharmacy consultation(14.7%).40.2% of the respondents hoped to learn medical knowledge.Conclusion Hefei city of Anhui province has a high proportion of urban family with standing drugs,the purchase way is regular,and when the disease,first of all to eat home standing drugs,medical treatment prefers large hospital,most think most intravenous infusion is not necessary;They want to understand the medical knowledge,and hope to establish consultation channels.

BACKGROUND:The in vivo degradation process of chitosan/hydroxyapatite composite porous scaffolds is not very clear. Research on the effects of rat osteoblasts and degradation products is less.
OBJECTIVE:To analyze the biocompatiblity of rat osteoblasts with degradation products of chitosan/hydroxyapatite composite porous scaffolds.
METHODS:The second generation of cultured rat osteoblasts were respectively cultured in the extract of degradation products of chitosan/hydroxyapatite composite scaffolds (experimental group) and Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum (control group). At 2, 4, 6, 8, 10 days of culture, cel counting was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Alkaline phosphatase activity was measured by the recommended method of determination of the Federation, and total protein was determined by BCA method.
RESULTS AND CONCLUSION:The proliferation speed, alkaline phosphatase activity, total cel ular protein synthesis and ratio of alkaline phosphatase to total protein in rat osteoblasts cultured in the experimental group were significantly higher than those in the control group (P<0.05). This experiment showed that the degradation products of chitosan/hydroxyapatite composite porous scaffolds cannot only promote rat osteoblast adhesion, growth and proliferation, but also enhance its ossification function, with good biocompatibility.

Objective To study the developmental process of the region of basal nuclei of postmortem fetuses by 3.0 T and 7.0 T MRI.Methods One hundred and thirty-one postmortem fetuses of 14 to 40 weeks of gestational age(GA)were scanned by 3.0 T MR,of which 11 fetuses of 14-27 weeks of GA were chosen and scanned by 7.0 T MR. The time when the structures in the region of basal nuclei could be detected and the changes of MR signal intensity were analyzed for MRI of different Tesla.Results On 3.0 T MRI.the dorsal thalamus could be delineated as early as 14 weeks of GA. The germinal matrix, caudate nucleus,and putamen could be visualized as early as 15 weeks of GA. The globus pallidus could be described as early as 18 weeks of GA.and the internal capsule and external capsule could be shown as early as 20 weeks of GA. The signal of the caudate nucleus during 15-30 weeks of GA was relatively hypointense on T1WI and hyperintense on T2WI.but during 31-40 weeks of GA, it was relatively hyperintense on T1WI and hypointense on T2WI. The putamen had a relatively high signal intensity on T1WI and low signal intemity on T1WI during 15-17 weeks of GA, and it appeared patchy during 18-25 weeks of GA,then it had a relatively low signal intensity on T1WI and high signal intensity on T2WI during 26-30 weeks of GA, and during 31-40 weeks of GA,its signal intensity was relatively high on T1WI and low on T2WI.The globus pallidus had a relatively high signal intensity on T1WI and low signal intensity on T2WI during 20-40 weeks of GA Compared to the 3.0 T MRI,the T2 images of 7.0 T MRl were more clear,and most structures in the region of basal nuclei could be clearly displayed as early as 16 weeks of GA.such as the germinal matrix,caudate nucleus,dorsal thalamus,putamen,globus pallidus,internal capsule,and external capsule.The claustrum could be delineated as early as 18 weeks of GA on 7.0 T MRI. Conclusions 3.0 T MRI could show the developmental process of the region of basal nuclei well,but the T2 images of 7.0 TMRl were comparatively better.

The roles of Indian hedgehog (Ihh) signaling pathway in the proliferation and apoptosis of precartilaginous stem cells (PSCs) were investigated.PSCs,labeled with fibroblast growth factor receptor 3 (FGFR-3),were isolated from neonatal rats by immanomagnetic separation.After identifi-cation with FGFR-3 and Col Ⅱ,the cells were incubated with different concentrations of cyclopamine (cyclo),the specific inhibitor of lhh signaling pathway.The morphologic changes of the cells were observed under the inverted phase contrast microscope.The mRNA expression levels of Ibh,para-thyroid hormonerelated peptide (PTHrP),protein Patched (Ptch),Bcl-2 and p21 were detected by RT-PCR.The protein expression levels of Ihh and Ptch were measured by Western blot.MTT assay was used to examine the effects of cyclo on proliferation of PSCs.Apoptosis rate of PSCs was exam-ined by Annexin V/PI assay of flow cytometric analyses.After PSCs were incubated with cyclo,ob-vious morphologic changes were observed as compared with the control group.The mRNA expres-sion levels of PTHrP,Ptch and Bcl-2 were decreased to varying degrees in a cyclo dose-dependent manner.However,the expression levels of lhh and p21 mRNA were increased.The protein expres-sion of Ptch and Ihh had the same change as the mRNA expression.Meanwhile,cyclo could obvi-ously inhibit the proliferation and promote the apoptosis of PSCs.The results indicated that Ihh sig-naling pathway plays an important role in regulating the proliferation and apoptosis of PSCs,which is probably mediated by Bcl-2 and p21.

Objective:To investigate the effect of ureteral decellularized matrix (UDM) coating on the differentiation of adipose stem cells.Methods:From January 2018 to October 2019, UDM was prepared by perfusion method. H&E staining, DAPI staining, and DNA quantification were used to assessment the residues of cellular component in UDM and normal ureter; Masson′s trichrome staining and collagen quantification evaluated the collagen retention in UDM and normal ureter; the distribution and content of glycosaminoglycan in UDM and normal ureter were analyzed through Alcian Blue staining and glycosaminoglycan quantification. Canine adipose mesenchymal stem cells (cADMSCs) were isolated and cultured and identified by flow cytometry. The UDM was digested by pepsin enzyme to prepare the decellularized matrix coating as the experimental group. Type Ⅰ rat tail collagen coating was used as a control group. The cADMSCs were seeded on different coatings, and the differentiation of the cADMSCs was detected by immunofluorescence staining and Western Blot.Results:H&E and DAPI staining showed that the nuclear residue was not observed in the UDM. The DNA quantification demonstrated that the DNA content in UDM [(38.87±3.40) ng/mg] was significantly lower than that in the normal ureter group [(1 694.63±169.83) ng/mg, P<0.05]. Masson′s trichrome staining and collagen quantification confirmed that the collagen content in UDM [(265.89 ± 16.40) μg/mg] was no significantly different from the normal ureter group [(288.73 ± 16.32) μg/mg, P>0.05]. Alcian Blue staining showed the distribution of glycosaminoglycan in the UDM, and glycosaminoglycan quantification suggested that the content of glycosaminoglycan in the UDM [(1.57 ± 0.19) μg/mg] was lower than that in the normal ureter group [(3.43 ± 0.12) μg/mg] ( P<0.05). Immunofluorescence staining and Western Blot confirmed that the expression of Alpha-smooth muscle Actin (α-SMA) in the experimental group (2.51 ± 0.27, 3.68 ± 0.33, 4.91 ± 0.45) was higher than that in the control group (0.97±0.09, 1.02 ± 0.10, 1.00 ± 0.11) at 3 d, 7 d, 10 d ( P<0.05). The expression of α-SMA in the experimental group increased gradually with culture time ( P<0.05). While no changing of α-SMA expression in the control group was recordered. Conclusions:The prepared UDM removed the cellular components, and retained the collagen structure and bioactive components well; the ureter decellularized matrix coating could promote the differentiation of cADMSCs to smooth muscle cells.

Objective:To investigate the effect of tissue engineered bladder patch constructed by double-layer silk scaffold and adipose-derived stem cells (ADSCs) in the repair and reconstruction of bladder.Methods:This study was conducted from May 2020 to March 2021. The silk fibroin (SF) aqueous solution was obtained from silkworm cocoons, and a double-layer silk scaffold composed of silk fibroin film and silk fibroin sponge was further prepared. The rat ADSCs were isolated, cultured, and the ADSCs surface markers (CD29, CD90, CD45, CD106) were identified by flow cytometry. The ADSCs were planted on a double-layer silk scaffold to construct a tissue-engineered bladder patch. Thirty-six male SD rats were randomly divided into three groups: tissue engineered bladder patch group (SF-ADSCs group, n=15), double-layer silk scaffold group (SF group, n=15), control group ( n=6). The tissue engineered bladder patch (SF-ADSCs group) and double-layer silk scaffold (SF group) were wrapped on the omentum to promote vascularization. The vascularization was evaluated by HE and immunofluorescence staining. The wrapped tissue engineered bladder patch and double-layer silk scaffold were used to repair the defective bladder. In the control group (six rats), the incision was closed immediately after the bladder tissue fully exposed. At 4 weeks and 12 weeks after operation, the general morphology of bladder tissue and cystography were performed to evaluate the recovery of bladder morphology. After the graft was harvested, HE and Masson's trichrome staining and immunofluorescence staining were used to observe the regeneration of bladder wall tissue. Urodynamics was used to assess the recovery of bladder function at 12 weeks after operation. Results:The flow cytometry results confirmed that the isolated cells positively expressed CD29 and CD90, and there was no significant expression of CD45 and CD106. Gross observation and scanning electron microscope confirmed that the preparation of double-layer silk scaffold not only had a pore structure that was conducive to cell planting, but also had good toughness and was facilitated to surgical suture. The number (43.50±2.66) and area (0.73±0.03)% of vascular-like structures in the SF-ADSCs group after the omentum encapsulation was significantly higher than that in the SF group [(24.50±3.51), (0.55±0.05)%], and the difference was statistically significant ( P<0.05). At 4 weeks after bladder repair, the histological staining of the grafts in the SF-ADSCs and SF groups showed a large number of degraded fragments of double-layer silk scaffold. At 12 weeks, the morphology of the graft in the SF-ADSCs group showed uniform bladder morphology, which was similar to that of normal bladder tissue. Immunofluorescence staining showed that the continuous urothelial layer, abundant smooth muscle tissue, vascular structure and regenerated neurons could be observed in the SF-ADSCs group. Urodynamic test showed that the bladder maximum volume (0.74±0.03)ml and compliance (16.68±0.44)μl/cm H 2O in the SF-ADSCs group, which were better than that in the SF group [(0.47±0.05)ml, (14.89±0.37)μl/cm H 2O], but lower than that in the control group [(1.12±0.08)ml, (19.34±0.45)μl/cm H 2O], and the difference was statistically significant ( P<0.05). Conclusions:The tissue engineered bladder patch constructed with double-layer silk scaffolds and ADSCs could promote the morphological repair of bladder tissue, the regeneration of bladder wall structure and the recovery of bladder physiological function.

Objective:To prepare the whole bladder acellular matrix (BAM) using the self-designed perfusion decellularization system, and evaluate the feasibility of constructing the tissue engineering bladder with the adipose-derived stem cells (ADSCs).Methods:This study was conducted from October 2020 to April 2021. The self-designed perfusion decellularization system was used, and four different decellularization protocols (group A, group B, group C and group D) were formulated, according to the flow direction of the perfusate and the action time of different decellularization solutions. Among them, the urethral orifice of the bladder tissue was used as the outflow tract of the perfusion fluid in groups A and B. The top of the bladder was cut off and used as the outflow tract of the perfusion fluid in groups C and D. In groups A and C, 1% Triton X-100 was treated for 6 h, and 1% sodium dodecyl sulfate (SDS) was treated for 2 h. In groups B and D, 1% Triton X-100 was treated for 7 h, and 1% sodium dodecyl sulfate (SDS) was treated for 1 h. In addition, the tissue in the normal bladder group was directly obtained from the natural bladder tissue, which did not require perfusion, cryopreservation and thawing. The fast and efficient decellularization protocol was screened out through HE, DAPI, Masson trichrome and Alcian Blue staining and quantitative analyses to prepare the whole bladder scaffold. The prepared BAM was used as the scaffold material, and the ADSCs were used as the seeding cells to construct the tissue engineering bladder. HE and DAPI staining were used to observe the distribution of ADSCs on the BAM.Results:HE and DAPI staining showed that there was no obvious nuclear residue in the group C. Masson trichrome and Alcian Blue staining showed that the collagen structure and glycosaminoglycan were well preserved in the group C. There was no significant difference in bladder wall thickness between the group C and the normal bladder group [(975.44±158.62)μm vs.(1 064.49±168.52)μm, P > 0.05]. The DNA content in the group C [(43.59 ±4.59) ng/mg] was lower than that in the normal bladder group, group A, group B and group D [(532.50±26.69), (135.17±6.99), (182.49±13.69) and(84.00±4.38)ng/mg], and the difference was statistically significant ( P<0.05). The collagen content [(10.98 ± 0.29)μg/mg] and glycosaminoglycan content [(2.30±0.18)μg/mg] in group C were not significantly different with those in the normal bladder group [(11.69±0.49) and (2.36±0.09)μg/mg, P>0.05]. Scanning electron microscopy showed that a large number of pore structures could be observed on the surface of the prepared BAM in groups A-D and were facilitated to cell adhesion. The isolated and cultured ADSCs were identified by flow cytometry to confirm the positive expression of CD90 and CD29, and the negative expression of CD45 and CD106. Live/dead staining and CCK-8 detection confirmed that the prepared BAM in the group C had no cytotoxicity. HE and DAPI staining showed that a large number of ADSCs were distributed on the surface and inside of the tissue engineering bladder. Conclusions:The whole bladder shape BAM prepared by the self-designed perfusion decellularization system could be used as the scaffold material for bladder tissue engineering, and the constructed tissue engineering bladder could be used for bladder repair and reconstruction.

Objective:To determine the correlations of single nucleotide polymorphisms (SNPs) with atrial fibrillation (AF) in the Chinese Han population from the central plains.Methods:A total of 168 hospitalized patients,including 56 AF and 112 controls,were recruited in this case-control study.The clinical data were obtained from the medical records.All 5 SNPs,rs337711 in KCNN2,rs11264280 near KCNN3,rs17042171 near PITX2,rs6771157 and rs6795970 in SCN10A,were genotyped using amplification refractory mutation system-polymerase chain reaction or direct sequencing.The x2 test was used to compare categorical variables and preliminarily examine correlations between the genotype frequencies and AF.Subsequently,a logistic regression model was constructed to determine the associations between the SNPs and AF based on the above screened results.Odds ratios (ORs) and 95％ confidence interval (CI) were calculated to assess the strength of the correlations.Moreover,we downloaded the genotype data from the HapMap Project for linkage disequilibrium analysis ofrs17042171.Results:AF patients were likely to be of older age and longer left atrial diameter and had more coronary artery disease and higher hypertension compared with the control group (P＜0.05).Among the 5 SNPs,the frequency distribution of genotype AA for rs17042171 was significantly different between the AF and control groups (P＜0.05).After adjusting for several covariates,there was still a high risk ratio in patients with the AA genotype compared with the AC+CC genotype (OR:5.591,95％CI 2.176 to 14.365,P-B＜0.008).Similarly,stratification analysis on the AA genotype demonstrated significant differences between rs17042171 and persistent AF.However,there were not significant correlations between AF and the control groups for the other 4 SNPs (P＜0.05).Conclusion:Rs17042171,near PITX2 on chromosome 4q25,is associated with AF susceptibility in the Chinese Han population from the central plains,suggesting that this SNP can provide a new strategy for clinical diagnosis in AF patients.

Objective:To investigate the effect of different acquisition duration of brain image of 18F-florbetaben(18F-FBB)positron emission tomography(PET)on standardized uptake value(SUV).Methods:Eight subjects who underwent 18F-FBB PET examination in Chinese PLA General Hospital from May 2021 to June 2021 were selected,including 5 persons of healthy control and 3 patients with mild cognitive impairment(MCI).All subjects underwent 18F-FBB PET imaging,and the dynamic PET images of them on brains were continuously acquired for 20 min between 90 and 110 min after the 18F-FBB injection was injected as(3.7-5.5 MBq/kg).Under the situation that other reconstruction parameters did not change,the images were reconstructed at 0-1,0-3,0-5,0-10,0-15 and 0-20 min,respectively.The same of region of interest(ROI)ranges were delineated in bilateral frontal cortex,bilateral temporal cortex,bilateral parietal cortex,posterior cingulate gyrus and cerebellar cortex of each group of images.And then,the corresponding mean standardized uptake value(SUVmean)of each region was obtained.The differences of SUVmean values of different ROI values between each group of data images and the images of 0-20 min were compared and analyzed.Results:There were significant differences in SUVmean between the acquired images in 0-1,0-3,0-5 and 0-10 min and the acquired standard images of 0-20 min(t=-7.569--2.410,P<0.05),respectively.There were no significant differences in SUVmean between the acquired images of 0-15 min and the acquired standard images of 0-20 min in the bilateral frontal lobe,bilateral temporal lobe,bilateral parietal cortex and posterior cingulate gyrus(P>0.05),only there was significant difference in the cerebellar cortex area between them(t=-5.597,P<0.001).Conclusion:The results of 15 min can reach to the similar results of 20 min in acquiring images,which can shorten the time of examination,and enhance the degrees of comfort and cooperation of patients in examination.It has clinical application value.