Aim To establish ICR animal model with C6 glioma stem cells, to provide the ideal model for the further study of gli-oma stem cells in brain glioma model. Methods C6 glioma stem cell was cultured in vitro by suspension,and was identified with Nestin antibody. C6 stem cells of ICR mouse glioma model were used to investigate survival state and tumor volume in mice after the operation. HE staining and CD133 immunohistochemi-cal study were adopted to investigate the postoperative pathologi-cal changes in mice. Results The expression of Nestin was 96. 01% in C6 glioma stem cells, and Nestin was highly ex-pressed in the cultured C6 glioma stem cells. Mice were inocula-ted with tumor after loss of appetite, weight, behavior and slow, sluggish reaction. Tumor volume at day 21 after modeling was (9. 77 ± 6. 58) mm3 . After HE staining, the model showed the invasive growth, tumor cell shrinkage and derangement. Immu-nohistochemical CD133 staining revealed that tumor cytoplasm color was brown. Conclusion Glioma model can be established based on glioma stem cells into a high rate of tumor, the tumor cycle is short, which can be used as an ideal model for glioma.

BACKGROUND:The cell-sheet technology, based on a temperature-responsive culture, has been drawing more and more attention;however, the temperature-responsive culture dish is quite expensive. Therefore, it is imperative to develop a substitutive technique.OBJECTIVE: To study the feasibility of cell-sheet ulturing using common culture dish, and investigate the chondrogenesis of the cell sheet. METHODS: A piece of nasal septal cartilage was adopted from a patient with deviation of nasal septum to extract primary chondrocytes that were then cultured and amplified. The passage 3 chondrocytes were used to construct ell sheets. Monolayer cell sheet was formed by intensive culturing and allowing the extracellular matrix secretion. Bilayer cell sheet was constructed by seeding passage 2 chondrocytes on the monolayer cell sheet. The cell sheets were harvested using cell scraper, their properties were investigated prior to plantation into nude mice to construct the tissue-engineered cartilage. RESULTS AND CONCLUSION: Both bilayer and monolayer cell sheets with soft tremellose structures showed no significant difference through naked eyes. The newly harvested cell sheets appeared to have good fluidity and gelation. Eight weeks after mplantation into the nude mice, mature cartilage blocks were obtained. Histologically, the cell sheets were thin films composed by layered chondrocytes and extracellular matrix. Glycosaminoglycan formation and type Ⅱ collagen expressions were observed in the cell sheets cultured in vitro. The explanted samples exhibited ature cartilaginous tissue at 8 weeks after implantation. Biochemical analysis showed that the DNA contents of the neocartilages were higher than those of native human costal cartilage, while the contents of glycosaminoglycan and hydroxyproline were similar to native human nasal septal cartilage. To conclude, the hondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis.

BACKGROUND:To seek for ideal scaffold materials is still an important task for cartilage tissue engineering.OBJECTIVE:To investigate the application of the AviteneTM microfibrillar collagen hemostat sponge in cartilage tissue engineering.METHODS:Rabbit auricular cartilage was harvested via surgical operation,and primary chondrocytes were isolated and amplified.Microfibrillar collagen hemostat sponge was cut into small bricks.The passage 2 chondrocytes were suspended and seeded onto the spongy bricks.After 1 week of in vitro culture,the constructs were then implanted into nude mice.After 8 weeks,the specimens were collected and evaluated using gross,histological and immunohistochamical observation.RESULTS AND CONCLUSION:During the cell seeding,the scaffold maintained its dimensions.No shrinkage was observed when the cell suspension was added.There was no considerable change in dimensions during the 1-week in vitro culture and at 8 weeks after implantation in nude mice.At 8 weeks post-implantation,mature cartilage blocks were harvested,which were white,translucent,and flexible.Histologically,the constructs appeared to have typical mature cartilaginous tissues,with robust extracellular matrix secretion,in which the microfibrillar collagen was incompletely degraded.We conclude that the microfibrillar collagen is a favorable scaffold material for cartilage tissue engineering.

Objective]To study the feasibility of Cartilage engineering using fibrin gel and chondrocyte cell sheets.[Methods]rabbit auricular chondrocytes were isolated and cultured to form cell sheets in flasks. The cell sheets were harvested using cell scrapers,and cut into fragments. The two precursor solutions of Fibrin gel were used to suspend the cell sheet fragments and isolated chondrocytes,and then added into the wells of a 48-well plate to form Gelatinous chondroid disc constructs. After in vitro culture, the constructs were implanted into nude mice. After 8 weeks,the constructs were harvested,and the specimens were evaluated using grossly observing, histological and immunohistochemical observation. [Results]Mature cartilage discs were obtained. The histomorphology of the explanted discs appeared non-uniform cartilaginous tissue comprise of regenerated cartilage islands with different size and irregular shape. Immunohistochemistry staining demonstrated that type II collagen highly expressed in the ECM of the cartilage islands. In 1 of the 8 discs,partial ossification was observed.[Conclusion]Fibrin gel is a favourable carrier. Artificial cartilage with stereochemical structure was constructed via combining the fibrin gel and chondrocyte cell sheets.

Objective:To explore the clinical effects of penile ventral Z-shaped cross flap and penile dorsal pedicled transfer flap in penoplasty for concealed penis.Methods:From January 2017 to June 2019, the data of 151 patients with concealed penis admitted to our hospital was retrospectively reviewed. The patients were divided into 2 groups according to the surgical method. 69 cases were treated with penis ventral Z-shaped cross flap to form penis and 82 cases were treated with dorsal penis pedicled flaps to form the penis. In Z-shaped flap group, the penis length of 33 patients with tight scrotum was (3.06±0.25)cm before surgery and the penis length of 36 patients with relaxed scrotum was (2.99±0.28) cm before surgery. In flap with transfer group, the penis length of 39 patients with tight scrotum was (3.04±0.30)cm before surgery and the penis of 43 patients with relaxed scrotum was (3.04±0.24)cm before surgery. The length of the penis after surgery and incidence of postoperative complications were compared between Z-shaped flap group and flap with transfer group. Common complications included penile body retraction, foreskin edema, foreskin stenosis and penile wound splitting.Results:151 patients were followed up for 6-12 months, and all patients were satisfied with penis appearance. There was no penile necrosis or urinary fistula. In Z-shaped flap cross group, the penis length of 33 patients with tight scrotum extended (2.47±0.22)cm after surgery.The penis length of 36 patients with scrotum relaxation extended (2.61±0.27)cm after surgery, 39 patients was adopted the penile dorsal pedicled transfer flap with scrotum tight had extended penis (2.90± 0.16)cm, which significantly different from the Z-shaped flap group( P<0.05). In flap with transfer group, 43 patients with relaxed scrotum extended (2.79±0.18)cm after surgery, which was significantly different from the Z-shaped flap group ( P<0.05). In Z-shaped flap group, 33 patients with scrotum tight, there were 2 cases of penile retraction, 1 case of stenosis of the foreskin, 2 cases of foreskin edema, 2 cases of penile wound rupture. In transfer flap group, of 39 patients with scrotum tight, there was 1 case of foreskin edema. The incidence of complications that adopted the penile dorsal pedicled transfer flap with scrotum tight was lower than those adopted penile ventral Z-shaped cross flap [2.56%(1/39) vs. 21.21%(7/33), P=0.033]. In transfer flap group, of the 43 patients with scrotum relaxation, there were 3 cases of penile retraction, 3 cases of foreskin stenosis, 2 cases of penile ventral foreskin edema, and 1 case of penile wound rupture. Z-shaped flap group: 36 patients was scrotum relaxation was 1 case of foreskin edema. The incidence of complications that adopted the penile dorsal pedicled transfer flap was higer than those adopted penile ventral Z-shaped flap [20.93%(9/43) vs. 2.78%(1/36), P =0.038]. Conclusions:In terms of children with tight scrotum or loose scrotum, the effect of the transfer flap method to extend the penis is better than that of the Z-shaped flap method. However, the transfer flap method has a low complication rate for children with tight scrotum, while the Z-shaped flap method has a low complication rate for children with loose scrotum.

Objective To investigate the effects of intense low frequency noise on expression of caspase -3 in bama pigs ,and observe the death of cochlea hair cell .Methods 8 bama pigs were randomly divided into a normal control group(2 pigs) and experimental groups(6 pigs) .Auditory brainstem responses(ABR) were tested before the experiment .The control group was the same as the experimental groups except noise exposure .The experimental group was randomly divided into immediate group ,36 h group ,and 84 h group(2 pigs per group) .They were ex-posed to intense low frequency noise at 142 dB of 50 Hz for 5 min according to the three time points .ABR were test-ed again before the cochlea were collected at different time points .The expression of caspase -3 was studied through immunohistochemistry and immunofluorescence ,hair cell loss rate by counting to determine cell death .Results The average ABR threshold of 8 bama pigs(16 ears) before the exposure was 61 .25 ± 10 .72 dB nHL .ABR thresh-olds were not elicited after exposure to the noise .Different parts of bama pigs showed positive expression of caspase -3 in two ways .As time prolonged after exposure ,the positive expression of caspase -3 gradually weakened .The im-mediate group and 36 h group ,compared with the control group ,showed the apparent misplace of three out hair cells in the arrangement and levels .The 84 h group through immunofluorescence lost out hair cells ,and only inner hair cells were visible .Conclusion ABR thresholds were elevated after noise exposure .The procedure of hair cell nucleus damage and caspase-3 expression is different ,and the noise can induce opening apoptotic program of spiral ganglion .

Objective:To compare the advantages and disadvantages of laparoscopic robot-assisted transmesenteric approach and retrocolic approach disconnected pyeloplasty in the treatment of children with hydronephrosis.Methods:From October 2020 to March 2021, 19 children with hydronephrosis were divided into two groups: intra-renal type and extra-renal type. Among them, 15 were males and 4 were females. The average age of the patients was 3.5 years old (0.2 years old to 16.8 years old), and the average weight was 18.4 kg (5.5 kg to 67.0 kg). The average ERPF of affected kidney before surgery was 35.4%(23.0%-49.8%). All of them were treated with laparoscopic robot-assisted transmesenteric approach and retrocolic approach disconnected pyeloplasty. The operation was performed in accordance with the standard surgical procedures of the guidelines. After the insertion of the trocar, the children in the transmesenteric group were exposed to the renal pelvis by incising the colonic mesangium into the retroperitoneal space, while in the retrocolic group, the peritoneum was cut into the retroperitoneal space to expose the renal pelvis. After that, the steps of incision, cutting, tube placement, and suture of the renal pelvis and ureter were the same in the two groups. Among the 10 cases of the extrarenal type, 6 cases were in the transmesenteric group and 4 cases were in the retrocolic group; among the 9 cases of the intrarenal type, 5 cases were in the transmesenteric group and 4 were in the retrocolic group. There was no statistically significant difference in age, weight, and renal function of the affected side before operation in different surgical approach groups ( P>0.05). The operation time, intraoperative anastomosis time, intraoperative blood loss and postoperative hospital stay were recorded and compared. There was no statistical difference in the age, weight, and renal function of the affected side before the operation. Results:19 cases were followed up for 6 months, no complications such as fever or wound infection occurred. The operation was successfully completed in all patients, no patients were transferred to open surgery, and the hydronephrosis was significantly reduced. Symptoms disappeared in both groups. Of the 19 children. In children with extrarenal type, the operation time of the transmesenteric group and the retrocolic group were (108.8±15.5) min and (132.8±7.6) min, and the intraoperative anastomosis time was (40.7±6.1) min and (51.5±5.5)min, the estimated intraoperative blood loss was (9.5±2.1) ml and (9.3±0.8) ml, respectively, and the postoperative hospital stay was (9.0±1.6) d and (9.3±2.9) d. The operation time and the difference of intraoperative anastomosis time was statistically significant ( P<0.05). In children with intrarenal type, the operation time of the transmesenteric group and the retrocolic group were (136.6±7.9) min and (116.5±13.5) min, and the intraoperative anastomosis time was (52.8±6.9) min and (40.8±6.2), min, the estimated blood loss during the operation was (11.4±2.3) ml and (10.5±0.9) ml, and the postoperative hospital stay was (8.8±1.7) d and (8.0±1.6) d. The operation time and The difference of intraoperative anastomosis time was statistically significant ( P<0.05). The 19 cases were followed up for 6 months, and there was no complications such as fever or wound infection. The volume of hydronephrosis was significantly reduced compared with that before operation, and the renal blood perfusion increased compared with that before operation. The difference was statistically significant ( P<0.05). Conclusion:In terms of shortening the operation time and suture time, for laparoscopic robot-assisted transmesenteric approach and retrocolic approach disconnected pyeloplasty in the treatment of children with hydronephrosis, the transtransmesenteric approach is more advantageous in the treatment of extrarenal hydronephrosis, while the retrocolic approach is more advantageous in the treatment of intrarenal hydronephrosis.

In this paper, the research has been conducted by the Microsoft kinect for windows v2 for obtaining the walking trajectory data from hemiplegic patients, based on which we achieved automatic identification of the hemiplegic gait and sorted the significance of identified features. First of all, the experimental group and two control groups were set up in the study. The three groups of subjects respectively completed the prescribed standard movements according to the requirements. The walking track data of the subjects were obtained straightaway by Kinect, from which the gait identification features were extracted: the moving range of pace, stride and center of mass (up and down/left and right). Then, the bayesian classification algorithm was utilized to classify the sample set of these features so as to automatically recognize the hemiplegia gait. Finally, the random forest algorithm was used to identify the significance of each feature, providing references for the diagnose of disease by ranking the importance of each feature. This thesis states that the accuracy of classification approach based on bayesian algorithm reaches 96%; the sequence of significance based on the random forest algorithm is step speed, stride, left-right moving distance of the center of mass, and up-down moving distance of the center of mass. The combination of step speed and stride, and the combination of step speed and center of mass moving distance are important reference for analyzing and diagnosing of the hemiplegia gait. The results may provide creative mind and new references for the intelligent diagnosis of hemiplegia gait.
Algorithms ; Bayes Theorem ; Gait ; Gait Analysis ; methods ; Gait Disorders, Neurologic ; diagnosis ; Hemiplegia ; complications ; Humans ; Walking

Objective: To explore the utility of pharyngeal pH monitoring which positive standard is Ryan index in diagnosis of laryngopharyngeal reflux disease. Methods: In a retrospective study, clinical data of 590 patients who had symptoms laryngopharyngeal reflux disease from February 2016 to March 2017 were analyzed. All patients were received electronic laryngoscopy, assessment of reflux symptom index(RSI) and reflux finding score(RFS), and pharyngeal pH monitoring. SPSS 19.0 software was used to analyze the date. Results: There were 94 patients whose Ryan index were positive(15.93%). Among the 94 patients, 70 were positive during upright, 12 during supine and 12 during both upright and supine. There were 40 patients(6.78%)with pH decline events related to symptoms, while those Ryan index were normal. There were 536(90.85%), 417(70.68%), 233(39.49%) and 117(19.83%) patients with pH<6.5, pH<6.0, pH<5.5 and pH<5.0 events respectively. The positive rate of RSI, RFS, RSI and RFS, RSI or RFS were 44.24%, 16.78%, 7.12%, 53.90% respectively. The RFS score in Ryan index positive group was higher than that in Ryan index negative group[(8.2±2.4) vs (4.0±2.9), u=5.424, P<0.05], while the RSI score in Ryan index positive group was not statistically different from that in Ryan index negative group[(11.3±6.2) vs (12.7±5.8), t=1.247, P=0.167]. Conclusions Pharyngeal pH monitoring is an objective and non-invasive method which can reflect laryngopharyngeal reflux directly. However, with the Ryan index as a criterion for the diagnosis of laryngopharyngeal reflux disease, partial patients may be missed. Further studies are needed to obtain more accurate and objective laryngopharyngeal pH statistical index for diagnosis of laryngopharyngeal reflux disease.