Objective The prestin,a motor protein responsible for outer hair cell(OHC)electromotility,is expressed on the OHC surface.Previous experiments revealed that OHC electromotility and its associated nonlinear capacitance was mainly located at the OHC lateral wall and was absent at the apical cuticular plate and the basal nucleus pole.Immunofluorescent staining for prestin failed to demonstrate the prestin expression at the OHC basal ends in whole-mount preparation of the organ of Corti.The aim of this study is to investigate the distribution of prestin at OHC.Methods In this experiment,the localization of prestin protein in single dissociated OHCs from cochlea of normal mouse,rat and guinea pig,were examined by immunofluorescent staining and confocal microscopy.Results We found that prestin was uniformly expressed on the OHC basolateral surface,including its basal pole.No staining was observed on the cuticular plate and stereocilia.The OHC lateral wall had a trilaminate organization and was composed of the plasma membrane,cortical lattice,and subsurface cisternae.By with co-staining with a membrane marker di-8-ANEPPS,prestin-labeling was found locating at the outer layer of the OHC lateral wall.Further separating the plasma membrane from the underlying subsurface cisternae,using a hypotonic extracellular solution,prestin-labeling was shown locating at the plasma membrane instead of the subsurface cisternae.Conclusion The data revealed that prestin is expressed in the plasma membrane on the whole OHC basolateral surface.

Safety is a critical link restricting the development of TCM injections.Hospital centralized monitoring is of importance in the safety evaluation of TCM injections.However,the results of centralized monitoring studies usually deviated from the actual situation because of problems in design and process quality.Our research team have completed several projects over centralized monitoring in recent years.Based on the previous research experience,this paper mainly discussed the current statuation,the significance and methods for standard process of hospital centralized monitoring in regard to TCM injections,in order to improve the quality of centralized monitoring studies and provide technique support for recognizing the safety of TCM injections.

According to the related requirements of safety reevaluation of TCM injections,hospital centralized monitoring of the safety of TCM injection was consecutively pressed ahead.Centralized security monitoring was a large-scale real world research involving numerous data acquisition,transmission,verification and analysis.The quality of data acquisition and data verification directly affected the authenticity and reliability of research data and results.Therefore,data management played a significant role in the safety monitoring of TCM injections.Based on the analysis of previous researches,this paper discussed the implementation content,links and the frequent problems and corresponding solutions of data management during the safety monitoring of TCM injections,combining with the experience in the implementation of projects.

The implementation process management plays a important role for the quality of centralized safety monitoring study of post-marketing Chinese medical injections.One of the critical link is hospital.Currently,there is no principle or specification for hospital choosing,number of monitoring points,data collection responsible part and forms of data collection as well.These issues caused uneven quality,huge differences in results and questioned conclusion.Based on the previous research experience,this paper focused on discussing the link of hospital where research data may be influenced,and proposed some suggestions.

BACKGROUND: It has been successful to repair articular cartilage defects by using solid carrier as cytoskeleton. We tried to transplant liquid or gel carrier materials combined cells into the body of animals, and investigated its feasibility.OBJECTIVE: To investigate the feasibility of homo-transplatation with liquid or gel carrier materials of Pluronic F-127-recombinant human bone morphogenetic protein-2 (rhBMP-2) engineered chondrocytes for the repair of full-thickness rabbit articular cartilage defect.DESIGN: A controlled experiment.SETTINGS: Department of Orthopaedics, Weihai Municipal Hospital;Shandong Institute of Orthopaedics and Traumaology.MATERIALS: The experiments were carried out in the laboratory of Shandong Institute of Orthopaedics and Traumaology from November 2001 to September 2003. Thirty-six healthy adult New Zealand rabbits of 2.5-4.5 kg, either male or female, were divided into four groups according to the method of random number table: Pluronic F-127-rhBMP-2 engineered chondrocytes group, Pluronic F-127-rhBMP group, Pluronic F127 engineered chondrocytes group and blank control group, with 9 rabbits in each group.METHODS: After grouping, the 36 rabbits were made into models of articular cartilage defects. Pluronic F-127-rhBMP-2 was used as a vector of chondrocytes which were obtained from New Zealand rabbits after cultured and amplified in vitro. The mixture of Pluronic F-127, Pluronic F-127-rhBMP-2 and cultured chondrocytes was transplanted into the defects of articular cartilage that had been made previously with φb3.5 mm drill.There was not any treatment in the blank control group. At 4, 8 and 12 weeks postoperatively, the repairing conditions of the defects were evaluated with gross observation and histological observation under light microscope and under electron microscope. The repaire quality was assessed blindly according to the Wakitani scoring standard.MAIN OUTCOME MEASURES: ① Healing of cartilage defects; ② Property and morphology of the chondrocytes, characteristics, number and arrangement of collagens in matrix.RESULTS: ① In the Pluronic F-127-rhBMP-2 engineered chondrocytes group, the transplanted chondrocytes could grow better than those in other groups, the defected areas were completely filled at 4 weeks. The regenerated tissues at 8 and 12 weeks had similar appearance with the surrounding normal cartilage tissue, but vague. Delimitation. The histological examination showed that transparent cartilages formed, and the defects were healed. ② Under electron microscope at 8 and 12 weeks, there were mature transparent cartilages in the repaired tissues, and there were irregularly arranged slight, even and non-periodical collagen Ⅱ in surrounding. In the blank control group, only fibrous repair was observed, the regenerated tissue lacked elasticity with rough surface. ③ Repairing quality score: The scores at each time point in the Pluronic F-127-rhBMP-2 engineered chondrocytes group were significantly different from those in the other groups.Those in the Pluronic F-127-rhBMP-2 engineered chondrocytes group and Pluronic F-127-rhBMP-2 group and Pluronic F-127 engineered chondrocytes group were significantly different from those in the blank control group [4 weeks: (3.93±1.91), (4.56±1.07), (4.78±1.09), (8.44±1.13) points:8 weeks: (2.80±1.45), (3.24±1.00), (3.33±1.00), (8.44±1.13) points; 12 weeks (2.22±1.10), (3.01±0.69), (3.00±0.71), (9.00±0.87) points, P ＜ 0.001],but there were no significant differences between the two groups (P ＞ 0.05).CONCLUSION: The mixture of Pluronic F-127-rhBMP-2 and cultured chondrocytes can repair successfully the cartilage defects of femoral condyle of rabbit knees by means of hyaline cartilage than simple application of Pluronic F-127-rhBMP-2 or Pluronic F-127 engineered chondrocytes.

Objective To evaluate the reporting item quality of randomized controlled trials of Chinese herbal medicine as an adjunctive therapy for advanced non-small cell lung cancer. Methods The terms including Chinese medicine, Chinese herbal medicine, randomized controlled trials, and non-small cell lung cancer were searched in Chinese and English databases by computer systems to collect relevant literatures. Based on the CONSORT 2010 (consolidated standards of reporting trials 2010), the reporting item quality of the abstracts and main text in eligible papers was evaluated. Results Ninety-five eligible studies were identified from 692 potential eligible articles. None reported all of abstract and main text items in CONSORT 2010. Only 4.12％could be identified as the randomized trial in the title. More than 40％of reports showed the scientific background or rationale in the abstract but not in the main text. Three (3.16％) eligible reports defined the primary or secondary outcome measures. None reported complete information of subjects throughout the clinical trial process. Results A total of 95 eligible papers were collected, of which 0 papers reported complete abstracts and text entries according to the CONSORT. The titles of 4.12％papers were identified as randomized trials. More than 40％of the papers only reported the study purpose or hypothesis in the abstract. 3.16％of the papers reported primary and secondary efficacy index entries, and 0 papers reported complete information of subjects from enrollment to included data analysis sets. Conclusion There is a serious problem of items shortage in the literatures of randomized controlled clinical trials on Chinese herbal medicine as an adjunctive therapy for advanced non-small cell lung cancer. The randomized controlled clinical trials of Chinese herbal medicine as an adjunctive therapy for advanced non-small cell lung cancer should be reported based on the CONSORT to improve the quality of trials.

Purpose: Erectile dysfunction (ED) is a common postoperative complication of pelvic surgery for which there is currently no effective treatment. This study investigated the therapeutic effects and potential mechanisms of adipose derived mesenchymal stem cells-derived mitochondria (ADSCs-mito) transplantation in a rat model of bilateral cavernous nerve injury (CNI) ED. Materials and Methods: We isolated mitochondria from ADSCs and tested their quality. In vivo, twenty male Sprague Dawley rats were randomly divided into four groups: sham operation group and CNI groups that received intracavernous injection of either phosphate buffer solution, ADSCs-mito or ADSCs. Two weeks after therapy, the erectile function of the rats was evaluated and the penile tissues were harvested for histologic analysis and western blotting. In vitro, the apoptosis rate, reactive oxygen species (ROS), mitochondria derived active oxygen (mtROS) and adenosine triphosphate (ATP) levels were detected in corpus cavernosum smooth muscle cells (CCSMCs) after the incubation with ADSCs-mito. In addition, intercellular mitochondrial transfer was visualized by co-culture of ADSCs and CCSMCs. Results: The ADSCs, ADSCs-mito and CCSMCs were isolated and identified successfully. ADSCs-mito transplantation notably restored the erectile function and smooth muscle content of CNI ED rats. Moreover, the levels of ROS, mtROS and cleaved-caspase 3 were reduced and the levels of superoxide dismutase and ATP were increased after ADSCs-mito transplantation. In CNI ED rats, the mitochondrial structure of cells in penile tissues was destroyed. ADSCs could transfer its own mitochondria to CCSMCs. Pre-treatment with ADSCs-mito could significantly decrease apoptosis rate, ROS levels and mtROS levels as well as restore the ATP level in CCSMCs. Conclusions ADSCs-mito transplantation significantly ameliorated ED induced by CNI, with similar potency to ADSCs treatment. The ADSCs-mito might exert their effects via anti-oxidative stress, anti-apoptosis and modulating energy metabolism of CCSMCs. Mitochondrial transplantation should be a promising therapeutic method for treating CNI ED in the future.

OBJECTIVE: To explore strategies of prenatal genetic testing for fetuses featuring abnormal skeletal development. METHODS: Clinical data of 17 fetuses with skeletal dysplasia was collected. The results of genetic testing and outcome of pregnancy were analyzed. RESULTS: For 12 fetuses, the femur-to-foot length ratio was less than 0.9. Thirteen fetuses had a positive finding by genetic testing. One fetus was diagnosed with chromosomal aneuploidy, three were diagnosed with microdeletion/microduplications, and nine were diagnosed with hereditary bone diseases due to pathological variants of FGFR3, COL1A2, GPX4 or ALPL genes. CONCLUSION For fetuses with skeletal dysplasia characterized by short femur, in addition to chromosomal karyotyping and microarray analysis, sequencing of FGFR3 and other bone disease-related genes can improve the diagnostic rate.
Bone Diseases, Developmental/genetics* ; Female ; Fetus/diagnostic imaging* ; Genetic Testing ; Humans ; Karyotyping ; Pregnancy ; Prenatal Diagnosis ; Receptor, Fibroblast Growth Factor, Type 3/genetics* ; Ultrasonography, Prenatal

OBJECTIVE: To study the correlation between dosage and curative effect of blood coagulation factor VIII in the prevention and treatment of haemophilia A in children and to determine the suitable dose for prevention of hemophilia in developing countries. METHODS: For different body weights of child patient, every time we used the same dosage of blood coagulation factor VIII (250 U each time, 3 times a week) and observed and recorded the number of hemorrhages in child patients. Then we compared the number of hemorrhages with children without treatment to determine the curative effect. According to the different body weights, we calculated the dosage of VIII factor of blood coagulation per kilogram (hereinafter referred to as the dose), and used Spearman correlation coefficient to study the correlation between dose and curative effect. RESULTS: The number of hemorrhages in 58 child patients before the treatment was 4.36 ± 1.78, while after the treatment was 2.22 ± 1.04 (t=7.91, P<0.001). The Spearman correlation coefficient of child patients of 5-10 U/kg was -0.421 (P=0.005); the Spearman correlation coefficient of child patients of 10-15 U/kg was -0.331 (P=0.030); the Spearman correlation coefficient of child patients over 15 U/kg was -0.16 (P=0.325). CONCLUSION Prevention and treatment can significantly reduce the times of hemorrhage in hemophilia patients.
Blood Coagulation ; Child ; Factor VIII ; administration & dosage ; therapeutic use ; Hemophilia A ; therapy ; Hemorrhage ; prevention & control ; Humans

Objective:To investigate the clinical symptoms and cone-beam CT (CBCT) imaging characteristics of temporomandibular joint osteoarthritis (TMJOA) with chewing side preference (CSP).Methods:One hundred patients with TMJOA diagnosed in the Department of Stomatology, General Hospital of the Chinese PLA from January 2018 to December 2020 were enrolled, including 32 males and 68 females, with an median age of 27.5 years (16-71 years). According to the habit of CSP, 100 cases were divided into 71 cases of TMJOA with CSP group and 29 cases of TMJOA without CSP group. The clinical symptoms were observed, including pain, TMJ sounds, limited mouth opening as well as the radiograph imaging changes of condylar bone. When analyzing the radiograph imaging changes of condylar, the cases with bilateral TMJ symptoms were excluded and the remaining cases were divided into symptomatic sides and asymptomatic sides with CSP or without CSP according to the symptoms of the chief complaint. SPSS 25.0 was used to analyze the statistical data. Age data did not conform to normal distribution so that median and quartile spacing were used for description, and Mann-Whitney U test was used for nonparametric test. Qualitative data such as gender, clinical symptoms and condylar lesion types were described by composition ratio and chi-square test was performed. Results:There was no statistical significance in age and gender of TMJOA patients in the group with or without CSP ( P>0.05). The incidence of pain in CSP group [83.1% (59/71)] was marginally higher than that in non-CSP group [65.5% (19/29)] but without statistical difference (χ2 =3.71, P=0.054). There was also no significant difference in TMJ sounds and limitation of mandibular movement between the two groups (χ2 =0.11, P=0.742; χ2 =0.48, P=0.489). Among all of joints, the most common types of TMJOA were articular flattening and shortening and erosion. CBCT showed that erosion [65.0% (130/200)], flattening and shortening [73.0% (146/200)], subcortical sclerosis [42.0% (84/200)], osteophyte [30.5% (61/200)] and subcortical cystic [15.5% (31/200)]. According to the different groups of chief complaint sides, intra-group comparisons show that the proportion of erosion in symptomatic sides of CSP group [80.0% (40/50)] was significantly higher than that in asymptomatic sides of CSP group [50.0% (25/50)] (χ2=9.89, P=0.002). Inter-group comparisons show that the proportion of condyle flattening and shortening in symptomatic sides of CSP group [84.0% (42/50)] was significantly higher than that in bilateral joint of non-CSP group (8/15) (χ2=8.81, P=0.032). There was no significant difference in the proportion of subcortical sclerosis, osteophyte and subcortical cystic between the group with or without CSP ( P>0.05). Conclusions:TMJOA patients with CSP may be more prone to clinical symptoms of pain and CBCT imaging changes of condyle erosion as well as flattening and shortening. CSP may be a promoting factor for the development of TMJOA.