The effect of large-conductance calcium-activated potassium channels on the migration of pericytes in the mice of senile cochlear stria vascularis / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate whether large conductance calcium-activated potassium channel (BK(Ca)) was involved in the migration of pericytes (PC) in the mice of senile cochlear stria vascularis capillaries PC. Methods: C57BL/6J mice were divided into 3-month (n=10) and 12-month groups (n=10). Auditory brainstem response (ABR) was used to test the hearing threshold of each group. The immunofluorescence was used to detect the expression changes of osteopontin (OPN) and β-BK(Ca) channels on cochlear stria vascularis PC. The morphological changes of perivascular cells in cochlea were observed by transmission electron microscope (TEM). Cell experiment: The PC, which were in the stria vascularis of the cochlea were primary cultured and identified. A cell senile model was made with D-gal. The appropriate intervention concentration of low galactose (D-gal) was determined by CCK8. β-galactosidase (SA-β-gal) staining was used to evaluate the cell decrept level. The change of BK(Ca) channels current on PC were recorded by whole cell patch clamp technique. The expression of BK(Ca) channels on PC was detected by immunofluorescence. The migration and invasion ability of two groups were detected by using Scratch test and Transwell. The levels of OPN and β-BK(Ca) channels were detected by Western blot. SPSS 22.0 software was used to analyze the data. Results: The ABR threshold in the 12-month group was higher than 3-month group (t=12.66, P<0.01). In the 12-month group, the expression of β-BK(Ca) channel was lower and the expression of OPN was increased (t=14.64, P<0.01; t=20.73, P<0.01). In TEM, cochlear stria vascularis PC were tightly connected to endothelial cells in 3-month group, while PC were loosely connected to endothelial cells or PC soma were separated from the capillary in 12-month group. Cell experiment: The positive rate of PC in the primary cultured cochlear stria vascularis is above 95%. Compared with the SA-β-gal stained cells in the control group, the positive rate of 15 mg/ml D-gal intervention PC was 85% (t=36.90, P<0.01). Whole cell patch clamp BK(Ca) channels current decreased in the D-gal group compared with the young group PC (t=12.18, P<0.05). The OPN expression in the senile group was higher than control group (t=16.30, P<0.01), while the β-BK(Ca) channels expression was decreased (t=11.98, P<0.01; t=15.72, P<0.05), and migration ability raised (t=7.91, P<0.01;t=7.59, P<0.01). After intervened of BK(Ca) channels specific blocker IBTX in the D-gal group, the expression of OPN and migration were increased (t=4.26, P<0.05; t=5.88, P<0.01; t=21.97, P<0.01). Conclusion: PC migration capacity were increased during the senile period, and the expression of β-BK(Ca) channel was decreased. The administration of IBTX, a specific blocker of BK(Ca) channel, at the cell level could increase the migration capacity, suggesting that BK(Ca) might be involved in the migration of PC in the stria vascularis of the aging cochlea.

Effect of microvascular pericytes of cochlear stria vascularis on endothelial cell permeability in C57BL/6J mice / 中华耳鼻咽喉头颈外科杂志

Objective: To study the changes in the permeability of the blood labyrinth barrier of the aging cochlea in mice, and to establish a non-contact co-culture model of endothelial cells (EC) and pericytes (PC) to furtherly investigate the cochlear stria vascularis microvascular pericytes impact on the permeability of endothelial cells. Methods: C57BL/6J mice were divided into two groups, three months old as young group, 12 months old as senile group. Cell experiment was divided into four groups, EC group, EC+PC co-culture group, D-gal+EC group and D-gal+EC+PC co-culture group. Auditory brainstem response (auditory brain response, ABR) was used to detect the auditory function of the two groups of mice. Evans blue staining was applied to detect the permeability of the cochlear blood labyrinth barrier of the two groups of mice. Transmission electron microscopy was used to observe the ultrastructure of blood labyrinth barrier endothelial cells, pericytes and tight junctions in the two groups of mice. Immunohistochemistry was used to detect the expression levels of tight junction proteins in the stria vascularis of the cochlea of the two groups of mice. Transwell chamber was used to detect the permeability of endothelial cells. Western blot and immunofluorescence technology were used to detect the expression level of tight junction protein on endothelial cells. SPSS 20.0 software was used to analyze the data. Results: Compared with the young group, the ABR threshold of the aging group was significantly increased, the latency of wave I was prolonged (t=10.25, P<0.01;t=5.61, P<0.05), the permeability of the cochlear blood labyrinth barrier was increased and the expression of tight junction protein on the vascular stria was decreased (P<0.05). The cochlear ultrastructure showed that the cochlear vascular stria microvascular lumen was deformed, the basement membrane thickened and the tight junction gap between endothelium enlarged. The positive rate of ECs and PCs in primary culture was more than 95%. The cells induced by 15 g/L D-gal were determined to be senescent cells. Compared with EC group, the expression of tight junction protein in endothelial cells of D-gal+EC group decreased（t=7.42，P<0.01；t=13.19，P<0.05）and the permeability increased (t=11.17, P<0.01). In the co-culture group, the expression of tight junction protein between endothelial cells in EC+PC co-culture group and D-gal+EC+PC co-culture group increased and the permeability decreased. Conclusions: In aging mice, the permeability of cochlear blood labyrinth barrier will increase and the level of tight junction protein will decrease; in aging state, cochlear vascular stria microvascular pericytes may affect endothelial cell permeability by regulating the expression of tight junction protein.

The efficacy of green surgery pathway on senile hip fractures / 中华骨科杂志

To evaluate the efficacy of green surgery pathway on senile hip fractures. Methods The green surgery pathway was implement in senile (≥75 years) hip fracture cases in our department since March 2016. A historically con?trolled trial study was designed: 97 senile hip fracture patients admitted between March 2016 and March 2017 were included in the intervention group and 78 senile hip fracture patients admitted between March 2015 and March 2016 were included in the con?trol group. The patients’general information were recorded, including gender, age, type of fracture, internal medical conditions, surgery method, the American Association of Anesthesiologists classification of physical status (ASA) classification, et al. The hip function was evaluated by the Harris score system at one week,6 months and one year after surgery. The waiting time for operation, operation time, length of hospital stay, complications during hospitalization and hip Harris score were compared between the two groups. Results For the baseline data between the two groups in terms of gender, age, fracture type, surgery method, ASA classi?fication, and combined underlying disease, the differences were not statistically significant. The waiting time for operation of the green surgery pathway group (36.3±7.8 h) was significantly shorter than that in the control group (46.9±11.4 h, t=6.995, P=0.000). The operation time of the green surgery pathway group was 45.4±17.5 min, and that of the control group was 43.8±20.8 min, the differences were not statistically significant. The length of hospital stay in the green surgery pathway group (6.8±2.4 d) was signifi?cantly shorter than that in the control group (9.5±2.8 d), the difference was statistically significant (t=6.866, P=0.000). In the green surgery pathway group, one patient developed acute myocardial infarction at 15 h after surgery and died of invalid rescue. In the control group, one patient developed acute cerebral hemorrhage 3 d after surgery and died of invalid rescue. The incidence of pul?monary infection, urinary tract infection, and acne in the green surgery pathway group were lower than that in the control group, the differences were statistically significant (χ2PI=5.081, PPI=0.024; χ2UI=6.841, PUI=0.009; χ2acne=11.768, Pcne=0.001). For the inci?dence of acute myocardial infarction, acute cerebrovascular accident, pulmonary embolism and deep vein thrombosis between the two groups, the differences were not statistically significant. The Harris score of hip in the green surgery pathway was higher than that in the control group (72.6±13.1 points vs 62.2±15.4 points, t=4.826, P=0.000) one week after the surgery, the difference was statistically significant. While the score was slightly higher than that of the control group in the half year (93.8±16.8 vs 90.5±14.7) and one year (94.1±18.3 vs 92.4±15.9) after surgery, but the differences were not statistically significant. The mortality rate of the green surgery pathway was slightly lower than that of the control group within one year after surgery (6.19% vs 8.97%), but the dif?ference was not statistically significant. Conclusion The implementation of green surgery pathway can reduce the occurrence of complications in bed, promote the early hip rehabilitation, effectively shorten the hospitalization time. It is worth further promotion because of its social benefits and health economic benefits.

The efficacy of green surgery pathway on senile hip fractures / 中华骨科杂志

Objective@#To evaluate the efficacy of green surgery pathway on senile hip fractures.@*Methods@#The green surgery pathway was implement in senile (≥75 years) hip fracture cases in our department since March 2016. A historically controlled trial study was designed: 97 senile hip fracture patients admitted between March 2016 and March 2017 were included in the intervention group and 78 senile hip fracture patients admitted between March 2015 and March 2016 were included in the control group. The patients’ general information were recorded, including gender, age, type of fracture, internal medical conditions, surgery method, the American Association of Anesthesiologists classification of physical status (ASA) classification, et al. The hip function was evaluated by the Harris score system at one week,6 months and one year after surgery. The waiting time for operation, operation time, length of hospital stay, complications during hospitalization and hip Harris score were compared between the two groups.@*Results@#For the baseline data between the two groups in terms of gender, age, fracture type, surgery method, ASA classification, and combined underlying disease, the differences were not statistically significant. The waiting time for operation of the green surgery pathway group (36.3±7.8 h) was significantly shorter than that in the control group (46.9±11.4 h, t=6.995, P=0.000). The operation time of the green surgery pathway group was 45.4±17.5 min, and that of the control group was 43.8±20.8 min, the differences were not statistically significant. The length of hospital stay in the green surgery pathway group (6.8±2.4 d) was significantly shorter than that in the control group (9.5±2.8 d), the difference was statistically significant (t=6.866, P=0.000). In the green surgery pathway group, one patient developed acute myocardial infarction at 15 h after surgery and died of invalid rescue. In the control group, one patient developed acute cerebral hemorrhage 3 d after surgery and died of invalid rescue. The incidence of pulmonary infection, urinary tract infection, and acne in the green surgery pathway group were lower than that in the control group, the differences were statistically significant (χ2PI=5.081, PPI=0.024; χ2UI=6.841, PUI=0.009; χ2acne=11.768, Pcne=0.001). For the incidence of acute myocardial infarction, acute cerebrovascular accident, pulmonary embolism and deep vein thrombosis between the two groups, the differences were not statistically significant. The Harris score of hip in the green surgery pathway was higher than that in the control group (72.6±13.1 points vs 62.2±15.4 points, t=4.826, P=0.000) one week after the surgery, the difference was statistically significant. While the score was slightly higher than that of the control group in the half year (93.8±16.8 vs 90.5±14.7) and one year (94.1±18.3 vs 92.4±15.9) after surgery, but the differences were not statistically significant. The mortality rate of the green surgery pathway was slightly lower than that of the control group within one year after surgery (6.19% vs 8.97%), but the difference was not statistically significant.@*Conclusion@#The implementation of green surgery pathway can reduce the occurrence of complications in bed, promote the early hip rehabilitation, effectively shorten the hospitalization time. It is worth further promotion because of its social benefits and health economic benefits.

Tissue-engineered scaffolds for articular cartilage repair / 中国组织工程研究

BACKGROUND:Articular cartilage repair has been a difficulty in the clinical setting, which is mainly treated with autologous or al ogeneic osteochondral grafts, and cartilage periosteum or periosteum grafts. However, the limited source, secondary lesion and immunological rejection force some researchers to search for a novel treatment strategy, cartilage tissue engineering, that is of great significance for cartilage regeneration and repair. OBJECTIVE:To investigate the tissue-engineered scaffolds for the repair of articular cartilage defects. METHODS:The first author searched the PubMed and WanFang databases for the articles addressing tissue-engineered cartilage for articular cartilage defects published between 1991 and 2015 using the keywords“articular cartilage defect, scaffold, tissue engineered cartilage”in English and Chinese, respectively. The irrelative and repetitive literatures were excluded. RESULTS AND CONCLUSION:Final y 48 eligible literatures were enrol ed based on the inclusion and exclusion criteria. Cartilage tissue engineering possesses the advantages of control ability, little damage to tissue itself, and biological repair of injured cartilage. Tissue-engineered scaffold material is a critical factor in tissue engineering construction;therefore, it should hold biodegradability and histocompatibility. The commonly used scaffold materials include natural macromolecule materials (col agen, silk fibroin and chitosan), and synthetic polymer materials (polylactic acid and tricalcium phosphate). It is necessary to prepare composite scaffolds with high bioactivity integrate advantages of each material. The tissue engineering is bound to be a hotspot in the field of articular cartilage repair.

Optimization of sperm alkaline single-cell gel electrophoresis / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures.</p><p><b>METHODS</b>Using alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method.</p><p><b>RESULTS</b>Three agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05).</p><p><b>CONCLUSION</b>The optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.</p>

Expression of hypoxia-inducible factor 2 alpha and vascular endothelial growth factor in chondrocytes of articular cartilages in human osteoarthritis / 中国组织工程研究

BACKGROUND:Studies have found that vascular endothelial growth factor and hypoxia inducible factor are involved in the development process of osteoarthritis, but their correlation is rarely reported. OBJECTIVE:To observe the expression and correlation of hypoxia inducible factor-2α and vascular endothelial growth factor in chondrocytes of articular cartilages in human osteoarthritis. METHODS: Articular cartilage specimens were colected from 50 patients with knee osteoarthritis undergoing total knee joint replacement. According to the joint Kelgren-Lawrance (K-L) X-ray grouping classification standard, there were 18 cases of K-LIII level and 32 cases of K-LIV level. Besides, articular cartilage specimens from 10 patients undergoing amputation for legs tumor or traffic accident served as control group. Hematoxylin-eosin staining, Safranin O-Fast Green staining and Mankin scoring were performed to observe and evaluate the histological characteristics of articular cartilages of each group, immunohistochemical staining was conducted to detect the expression of hypoxia inducible factor-2α and vascular endothelial growth factor in chondrocytes of articular cartilages, and their correlations were analyzed. RESULTS AND CONCLUSION:The Mankin score of K-LIV group was significantly higher than those of K-LIII group and control group. Immunohistochemical staining revealed that the number of chondrocytes with positive expression of hypoxia inducible factor-2α or vascular endothelial growth factor in K-LIV group was significantly higher than that in K-LIII group and control group (P < 0.05). The expression of hypoxia inducible factor-2α and vascular endothelial growth factor increased in chondrocytes of articular cartilages of osteoarthritis patients, and to up-regulate the expression of vascular endothelial growth factor may be the regulatory mechanism of hypoxia inducible factor-2αinthe pathogenesis of osteoarthritis.

Biological characteristics of different sources of mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Mesenchymal stem cels, underin vivo orin vitro specific induction conditions, can differentiate into the cartilage, muscle, tendons and so on. Clinical trials concerning mesenchymal stem cels mainly include tissue repair (such as bone, cartilage and joint repair) and treatment of heart, liver, spinal cord injury and nervous system diseases. OBJECTIVE:To compare the biological characteristics of mesenchymal stem cels from different sources. METHODS: PubMed and CNKI databases were retrieved for articles related to sources of mesenchymal stem cels and biological characteristics of mesenchymal stem cels published from 1987 to 2015. The retrieved articles were summarized and analyzed in the folowing aspects: cel surface marker, proliferation, differentiation, migration, and function, so as to explore the merits and demerits of mesenchymal stem cels from different sources. RESULTS AND CONCLUSION:A difference in the proliferation ability and surface markers is found between different sources of mesenchymal stem cels. Immunological competence of mesenchymal stem cels from different sources may be correlated with their activation status, species differences, tissue sources and culture conditions, resulting the immunological competence of mesenchymal stem cels from different sources is not exact the same. In-depth understanding of the factors and mechanisms by which influence the migration of mesenchymal stem cels from different sources can enhance the migration ability of different sources of mesenchymal stem cels, and increase their efficiency in wound healing, tissue repair and regeneration treatment.

Osteochondral autograft and allograft and tissue engineering materials for repair of articular cartilage injury / 中国组织工程研究

BACKGROUND:In recent years, repair of articular cartilage injury has become an important field in basic medical research. Because injured articular cartilage is difficult to repair, the repair of articular cartilage injury has become a difficult hotspot.

Local intra-articular injection of sodium hyaluronate delays articular cartilage degeneration after traumatic osteoarthritis / 中国组织工程研究

Abstract BACKGROUND: Sodium hyaluronate is an effective treatment for osteoarthritis, but the underlying mechanism remains unclear. There is evidence that abnormal expressions of matrix metaloproteinase (MMP)-1, -3 and -9 and tissue inhibitor of metaloproteinase (TIMP)-1 and -2 show great effects on osteoarthritis. OBJECTIVE: To assess the influence of intra-articular injection of sodium hyaluronate on expressions of MMPs-1, 3, 9 and tissue inhibitor of TIMPs-1, 2 in the rabbit cartilage after osteoarthritis. METHODS: Twenty-four mature New Zealand white rabbits were divided into normal control, model, and sodium hyaluronate groups. The model and sodium hyaluronate groups underwent unilateral anterior cruciate ligament transection, and rabbits in the sodium hyaluronate group received 0.3 mL of 1% sodium hyaluronate via intra-articular injection at 4 weeks after modeling, once a week for 5 weeks. At 11 weeks folowing surgery, the rabbits were kiled and the cartilage was harvested to extract total RNA. mRNA expressions of MMPs-1, 3, 9 and TIMPs-1, 2 in the cartilage were analyzed using real-time PCR for each group. RESULTS AND CONCLUSION:Compared with the model group, the range and extent of cartilage damage was reduced in the sodium hyaluronate group (P < 0.01), and Mankin scores were noticeably decreased (P < 0.05). In the cartilage, mRNA expressions of MMPs-1, 3, 9 were enhanced and mRNA expressions of TIMPs-1, 2 were down-regulated in the model group. However, the mRNA expression levels of MMPs-1, 3, 9 and TIMPs-1, 2 in the articular cartilage were not obviously changed in the sodium hyaluronate group. These results suggest that MMPs-1, 3, 9 and TIMPs-1, 2 are involved in the progression of osteoarthritis and the therapeutic mechanism of sodium hyaluronate is not realized through the down-regulation of their expressions during development of osteoarthritis. Sodium hyaluronate for treatment of osteoarthritis is a complex process and the underlying mechanisms require further investigation.

Vascular endothelial growth factor and hypoxia-inducible factor suppress chondrocyte apoptosis / 中国组织工程研究

BACKGROUND:Osteoarthritis is a joint disease that primarily affects the cartilage. With the changes of the extracelular matrix, chondrocytes appear to have apoptosis. Vascular endothelial growth factor plays an important role in promoting endothelial cel division and proliferation and inducing angiogenesis. Hypoxia-inducible factor is a celular transcription factor and produces different reactions due to the oxygen content. Vascular endothelial growth factor and hypoxia-inducible factor are focused on inhibiting chondrocyte apoptosis. OBJECTIVE:To elaborate the effects of vascular endothelial growth factor and hypoxia-inducible factors on chondrocyte apoptosis. METHODS: Recent literatures related to chondrocyte apoptosis were summarized and analyzed. During the process of osteoarthritis, changes in vascular endothelial growth factors in chondrocytes and regulatory effects of vascular endothelial growth factor and hypoxia-inducible factor on chondrocyte apoptosis were elaborated.

Homology and carbapenemase gene in Acinetobacter baumannii / 中南大学学报(医学版)

OBJECTIVE@#To study the antibiotic resistance evolution, homology, phenotypes and genotypes of carbapenemase in Acinetobacter baumannii from clinical isolates.@*METHODS@#A total of 72 strains of Acinetobacter baumannii were isolated from Xiangya Hospital of Central South University from March to May 2012. Antimicrobial susceptibility test was carried out by automatic microorganism clinical analytical system VITEK-II. The homology of the 72 strains was analyzed by enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). Modified Hodge test was used to screen carbapenemases of the strains. Carbapenemase genes blaOXA-23, blaOXA-40 and blaOXA-58 were also amplified and sequenced.@*RESULTS@#The 72 strains of Acinetobacter baumannii remained sensitive to cefoperazone/sulbactam (resistance rate 8.33%), followed by Amikacin. Otherwise, they were resistant to most of the antimicrobial agents (resistance rate more than 70%). The 72 strains were identified as 7 epidemic clones, A-G, by means of ERIC-PCR and the phylogenetic relationship among D, E, F and G was very close, suggesting a nosocomial infection possibility. Totally 56 strains produced carbapenemase; 61 strains were positive for carbapenemase gene blaOXA-23 and 1 strain positive for blaOXA-58. All strains were negative for carbapenemase gene blaOXA-40.@*CONCLUSION@#Acinetobacter baumannii strains isolated clinically are resistant to most of the antimicrobial agents and nosocomial infection had been observed. Most of the strains produce carbapenemase, among which, blaOXA-23 gene is the main carbapenemase gene. blaOXA-58 gene exists in the Acinetobacter baumannii isolates from Hunan Province.

Study on the characteristic of assimilating nitrogenous phosphorous fertilizer and the accumulation disciplinarian of total tanshinons of Salvia miltiorrhiza / 中国中药杂志

<p><b>OBJECTIVE</b>To confirm the amount of fertilizer and the ration of fertilizer.</p><p><b>METHOD</b>Through the experiment of planting Salvia miltiorrhiza in pot and in field were carried out.</p><p><b>RESULT</b>When N:P = 1:1, The production was 1.8 times more than the contrast group in plotting experiment, and in field the production of the highest fertilizer plot were 2.5 times more than the contrast and the higher and high fertilizer plot is 2.25 and 1.2 times respective were than those in the contrast group.</p><p><b>CONCLUSION</b>The proper ration of nitrogenous fertilizer and phosphorous fertilizer is 1:1. Nitrogen shows negative effects to the accumulation of tanshinon IIA, the more the nitrogen ous fertilizer, The less the content of tanshinon IIA. In contrast, phosphor ous fertilizer shows good effects on the accumulation of tanshinon IIA. Phosphorous fertilizer could alleviate the decline of the content of tanshinon IIA by using nitrogenous fertilizer. The accumulation peak of the tanshinon IIA emerge in the period of 150 d.</p>