Objective:To establish a matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) assay for the identification of common Salmonella serotypes and provide etiology evidence for the early precise treatment of salmonellosis. Methods:A total of 500 strains were collected from different regions and sources and five predominant Salmonella serotypes ( Salmonella Typhi , Salmonella Paratyphi A , Salmonella Typhimurium , Salmonella Enteritidis , and Salmonella Indiana) of each strain was identified by agglutination test and whole-genome sequencing. The protein complex of the strains was extracted by using optimized pretreatment method to establish the fingerprint database of peptides for each Salmonella serotype. The new serotyping assays were established by using different modules based on the mass spectra database. Additional 155 strains with specified serotypes and variant sources were used to test and evaluate the accuracy of the new typing assays. Results:Five MALDI-TOF MS databases were established, and two new serotyping assays were established via peptide fingerprint mapping/matching and machine learning of the neuronal convolutional network respectively based on the databases. The results showed that the fingerprint matching approach could quickly identify five common Salmonella serotypes in clinical practice compared with the machine learning method, the accuracy of fingerprint matching assay to identify five Salmonella serotypes reached 100.00% and the serotyping can be conducted within a short time (15-20 minutes) and had a good reproducibility, while the machine learning method could not completely identify these serotypes. Moreover the sensitivity and specificity of fingerprint matching assay were all 100.00% respectively, while they were only 82.23% and 95.81% for machine learning method. Conclusion:The established Salmonella serotyping assay based on MALDI-TOF MS in this study can easily, rapidly and accurately identify different serotypes of Salmonella.

Propofol,as a widely used intravenous anesthetic,has been found to have the potential of anti-tumor growth and metastasis in recent years.At present,the research on the mechanism of pro-pofol inhibiting tumor growth and metastasis is still relatively limited.In this paper,we summarize the recent studies on the role of propofol in inhibiting tumor growth and metastasis and its possible mo-lecular mechanisms,such as regulating specific sig-naling pathways or key molecules,and interacting with other molecules.At the same time,the appli-cation of propofol,including the optimization of ad-ministration route and dose,as well as the possibili-ty of being an adjunct therapy to radiotherapy and chemotherapy,was analyzed,and the recent stud-ies on the mechanism of propofol inhibiting tumor growth and metastasis were reviewed,providing a reference for further exploration of the application and research direction of propofol in clinical anti-tu-mor growth and metastasis therapy.

Guilu-Erxian Gum, a famous prescription of Traditional Chinese Medicine, which could warm the kidney and benefit essence, tonify qi and nourishing the blood. It's main function is to treat chronic kidney deficiency, sour waist and soft knees, lack of essence and blood, spermatorrhea and impotence. This prescription has been changed and expanded in the dosage, preparation and efficacy of Chinese medicine included in the classics. Pharmacological studies have shown that it can strengthen the bone, play the role of anti-tumor, relieve bone marrow suppression, improve sperm quality and relieve post-traumatic stress disorder. The modern clinical application of this priscriptionhas been greatly expanded. It can be used in the treatment of adverse reactions after chemotherapy, liver cirrhosis, hypoproteinemia, osteoporosis, osteoarthritis, AD and other diseases. It is often used in combination with other medicine or treatment methods, and could achieve good results.

ObjectiveTo explore the effect of Guilu Erxianjiao on improving reproductive injury in diabetic rats and its possible mechanism. MethodFifty-three SD male rats were randomly divided into 5 groups， with 1 group as the normal group and the other 4 groups as the modeling groups. Rats in the modeling groups were fed with a high-fat diet combined with 30 mg·kg^-1 streptozotocin （STZ） intraperitoneal injection to induce diabetes， with the random blood glucose >16.7 mmol·L^-1 for 3 consecutive times as the criteria for inclusion in the model of diabetic reproductive injury. The rats with diabetic reproductive injury were then randomly divided into a model group， a Guilu Erxianjiao group （2 g·kg^-1）， a Vitamin E group （0.03 g·kg^-1）， and a Wuzi Yanzong pill group （0.6 g·kg^-1） according to the blood glucose level. The rats were given the corresponding drug dose intragastric administration， once a day for 4 weeks， and their body weight and blood glucose were measured weekly. After 4 weeks， samples were collected for index determination. Morphological changes in testis and epididymis were observed by hematoxylin-eosin （HE） staining， and apoptosis of testis cells was observed by in situ end labeling （TUNEL） staining. Sperm concentration and motility were detected by the semen analyzer. Serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） were determined by enzyme-linked immunosorbent assay （ELISA）. The content of superoxide dismutase （SOD）， propylene glycol （MDA）， reactive oxygen species （ROS）， and glutathione peroxidase （GSH-Px） in the testicular tissue was determined by ELISA. The expressions of nuclear respiratory factor-2 （Nrf2）， heme oxygenase 1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in the testicular tissue were detected by Western blot. ResultAs compared with the normal group， testicular tissue atrophy， decreased spermatogenic tubules， epididymal wall hyperplasia， and lumen stenosis were observed in the model group. Sperm concentration and motility decreased （P<0.01）， and serum levels of T， FSH， and LH decreased （P<0.01） in the model group. The content of ROS and MDA in the testis increased （P<0.01）， while that of SOD and GSH-Px decreased （P<0.01） in the model group. The expression of Bax increased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 decreased （P<0.01） in the model group. As compared with the model group， the pathological changes in the testis and epididymis in the Guilu Erxianjiao group were improved to some extent. Sperm concentration and motility increased （P<0.05， P<0.01）. In the Guilu Erxianjiao group， serum levels of T and LH increased （P<0.05， P<0.01）， while FSH levels showed no significant difference. The content of ROS and MDA in the testis decreased （P<0.01）， while that of SOD and GSH-Px increased （P<0.01） in the Guilu Erxianjiao group. The expression of Bax decreased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01） in the Guilu Erxianjiao group. ConclusionGuilu Erxianjiao improves the reproductive injury and sperm quality of diabetic rats to a certain extent， and the mechanism may be related to the improvement of oxidative stress and anti-apoptosis.

Objective To investigate the application and the effects of individualized hollow resin plugs in the prevention of external auditory canal stenosis after canaloplasty of external auditory meatus for microtia patients . Methods The clinical data of 65 patients (68 ears) with microtia and aural atresia hospitalized in our department from January ,2006 to December ,2015 were summarized .Otoplasty of external ears and canaloplasty of external auditory meatus were operated on all patients under general anesthesia .Thigh flap transplantation was lined in the bony canal .One month later ,individualized hollow resin plugs were made and worn for 6 to 12 months ,and fol-lowed up for 1～2 years .Results The shapes of the reconstructed auricles were satisfactory .Sixty-four ears with wide external auditory canal openings were recorded in 68 ears .Preoperative hearing was 70 .45 ± 5 .5 dB HL ,and postoperative hearing was 55 .55 ± 5 .2 dB HL .Their hearing was not affected .External auditory canal restenosis occurred in 4 ears .Conclusion The individualized hollow resin plugs do not affect the hearing in use ,and can effec-tively prevent the restenosis of the external auditory canal after microtia reconstruction .

Objective To investigate the effect of intense low frequency noise on the different hearing functions and hair cell damage in bama pigs.Methods Thirteen bama pigs were randomly divided into a normal control group (3 pigs) and an experimental group (10 pigs).The pigs in experimental group were randomly divided into 50 Hz subgroup (5 pigs) and 70 Hz subgroup (5 pigs).The 50 Hz subgroup was exposed to intense low frequency noise at 167～170 dB SPL,50 Hz for 30 min,and the 70 Hz subgroup was exposed to intense low frequency noise at 164 dB SPL,70 Hz for 30 min.Auditory brainstem response(ABR) and distortion product otoacoustic emission (DPOAE) were performed before and after noise exposure.The cochlear were collected and the inner ear morphology changes were studied.Results Before the experiment,the ABR threshold of the pig was 20～50 dB SPL,and the average of 50 Hz group was 33.5±9.4 dB SPL,the average of the 70 Hz group was 34.0±4.6 dB SPL.The level of the above 3 000 Hz DPOAE could be elicited.But they were not elicited after the exposure to the noise.The inner ear structures were damaged.The DAPI showed hair cell missing.Conclusion ABR and DPOAE were elevated after noise exposure.The auditory system of bama pigs had irreversible damages.Hair cells damaged were given priority to with necrosis,and the higher the level of intense low frequency noise,the more serious the damage to the hair cell.

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.

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 observe the morphological changes on cochlear hair cells of rats in simulated weightlessness and inboard noise and to investigate the different changes in three turns of hair cells.Methods: Thirty-two healthy SD rats, all males, were randomly divided into four groups: control group, weightlessness group, noise group and weightlessness+noise groups (n=8).Then rats were exposed to-30° head down tilt as simulated weightlessness and inboard noise including steady-state noise which was (72±2) dB SPL and impulse noise up to 160 dB SPL in spaceship environment.The control group was kept in normal condition for 8 weeks.Bilateral auditory brainstem response (ABR) thresholds were tested before and after exposure respectively, and immunofluorescence staining and scanning electron microscopy (SEMs) of basilar membrane were applied after exposure.Results: ABR threshold shifts of each group were higher after exposure.There was difference between ABRs of the experiment groups before and after exposure (P<0.05).IF showed that the inner hair cells (IHCs) missing was the main damage in the basal turn of weightlessness group, the hair cells in the middle turn were swell and in the top turn, the hair cells were not clear.In noise group, the main loss happened in the outer hair cells (OHCs) of the outermost layer.In weightlessness+noise group, the nuclear missing in the basal turn was apparent, and mainly happened at the outermost layer.Meanwhile, the missing of hair cells in the middle turn and top turn was seen at the innermost layer.SEM showed that the cilia in the basal turn of weightlessness group were serious lodging, and occasional absence.Furthermore, the basal cilia in noise group became lodged and absent, and the other two turns were seriously missing.And in weightlessness+noise group, the cilia missing in the basal turn was apparently seen.The damage degree of the four groups: weightlessness+noise group>noise group>weightlessness group>control group and the damage degree of the four turns of hair cells: basal turn>mid turn>top turn.Conclusion: The rats exposed to the above environment for 2 weeks displayed obvious changes in cochlea morphology, and the weightlessness+noise group had the most obvious damage.