[A pedicle gastrocnemius muscle flap transposition for salvage of an exposed prosthesis and deep infection of knee joint after total knee arthroplasty].

Objective: To explore the effect of a delayed pedicle gastrocnemius muscle flap transposition for the treatment of a knee joint deep infection secondary to wound necrosis after total knee arthroplasty (TKA). Methods: The clinical data of 7 patients treated in Shanxi Provincial People's Hospital with a delayed pedicle gastrocnemius muscle flap transposition from December 2015 to September 2019 for wound necrosis, exposed prosthesis and deep infection of knee joint after TKA were analyzed retrospectively. Before the muscle flap transplantation, 5 of the patients had received at least one debridement but failed for relapse, and resulted in an exposed prostheses and infected knee joint. Four patients were positive in their wounds or joint exudates bacterial culture, while the other 3 patients were negative but only with an obvious purulent secretion. The radiographs in all of the patients had no signs of lucent peripheral to or sink of the prosthesis. Results: The patients were followed-up for a mean time of 16.5 months (7-39 months). The flap and skin graft survived uneventfully with no pain, sinus, fistula, edema, and hematoma occurred. The appearances of the legs were normal. Only 1 patient had a mild limp, and the others gained almost a normal gait. One of the patients recurred 5 months after the gastrocnemius muscle flap transposition, and a two-stage revision procedure was applied, that involving a prosthesis remove and vancomycin impregnated cement (4 grams of vancomycin powder mixed with forty grams of polymethylmethacrylate) spacer implanted, and a new prosthesis was re-implanted 6 months later. The assessment of Knee Society Score (KSS) graded as: 4 patients classified as excellence, 2 as fine, 1 as general. Conclusions: Delayed pedicle gastrocnemius muscle flap transposition is an effective method for the complication of wound necrosis, deep infection, prosthesis exposure after TKA. This protocol was appropriate for those who have experienced a comparative long time of infection while had no signs of prosthesis loosening, and with which the implant may be salvaged, the defect be closed, and the infection be eradicated.

[microRNA-16-5p targeted tetraspanin 15 gene to inhibit the proliferation, migration and invasion of osteosarcoma cell through phospoinositide 3-kinase/protein kinase B signaling pathway].

Objective: To study the effects of miR-16-5p on proliferation, migration and invasion of osteosarcoma cells and its mechanism. Methods: Quantitative polymerase chain reaction (qPCR) and Western blotting were used to detect the mRNA and protein expression of miR-16-5p and TSPAN15 in human normal osteoblasts hFOB 1.19 and osteosarcoma cells MG63, Saos2 and HOS. The miR-16-5p or si-TSPAN15 was transfected into MG63 cells to observe its role in cell proliferation, migration and invasion. Cell proliferation was measured with MTT assay, cell migration and invasion were examined by Transwell, and the protein expression of CyclinD1, matrix metalloproteinase 2 (MMP-2), MMP-9, tetraspanin 15 (TSPAN15), phospha-tidylinositol3-kinase(p-PI3K) and phospha-protein kinase B(p-AKT) were determined by using Western blotting. The starbase website prediction combined with dual luciferase gene reporter assay was performed to analyze the targeting relationship between miR-16-5p and TSPAN15. miR-16-5p and pcDNA-TSPAN1 were co-transfected to assess the effect of high expression of TSPAN15 on overexpression of miR-16-5p-induced proliferation, migration and invasion of MG63 cells. Data comparison between the two groups was performed by using t test. Results: Compared with hFOB 1.19 cells (1.00±0.12), the expression of miR-16-5p was significantly decreased in MG63, Saos2 and HOS cells (0.32±0.05, 0.40±0.04, 0.45±0.06, respectively)(F=156.204, P<0.05), and TSPAN15 mRNA and protein levels were greatly increased (F=71.718, 110.350, both P<0.05). Overexpression of miR-16-5p obviously reduced the expression of CyclinD1, MMP-2, MMP-9 protein, cell viability, cell migration and invasion (F=150.136,117.228, 154.971, 89.479, 98.373, 130.880, all P<0.05) in MG63 cells. Knockdown of TSPAN15 greatly reduced CyclinD1, MMP-2, MMP-9 protein levels, cell survival rate, cell migration, and invasion number (F=93.206, 107.030, 109.326, 115.625, 146.113, 139.300, all P<0.05). Overexpression of miR-16-5p markedly decreased the expression of p-PI3K and p-AKT protein in MG63 cells (F=156.755, 181.419, both P<0.05). miR-16-5p targeted to regulate the expression of TSPAN15. High expression of TSPAN15 partially reversed the inhibitory effect of miR-16-5p on TSPAN15, CyclinD1, MMP-2, MMP-9, p-PI3K, p-AKT protein expression, cell viability, cell migration number and invasion number in MG63 cells. Conclusion: miR-16-5p inhibits the proliferation, migration and invasion of osteosarcoma cells by targeting the TSPAN15 gene and regulating the PI3K/AKT signaling pathway.

[Loss of autophagy in condylar chondrocytes causes increased apoptosis rate in temporomandibular joint osteoarthritis of rats].

Objective: To observe the effect of autophagy of condylar chondrocytes on apoptosis in temporomandibular joint osteoarthritis (TMJOA) of rats. Methods: Fourty male 2-month-old SPF SD rats were equally divided into sham group (n=20) and experimental group (n=20). UAC metal prosthesis was cemented to the left incisors of maxilla and mandible of the rats in experimental group rats. After 8 weeks, all rats were sacrificed and the temporomandibular joint was taken. Two groups of rat condylar chondrocytes were extracted and cultured in vitro to the third generation. Immunofluorescence technique was used to detect the levels of collagen Ⅱ and matrix metalloproteinase-13 (MMP-13) in chondrocytes. The level of light chain-3 (LC-3), an autophagy marker of chondrocytes, was detected. Immunohistochemical technique was used to detect the level glycogenin-1, a glycogen formation marker of chondrocyte, was detected. The level of caspase-3, an apoptosis marker of chondrocyte, was also detected. Tunel technique was used to detect the apoptosis rate of the two groups at 72 h. Cracking cell extraction of total protein, Western-blotting (WB) technology to detect the levels of collagen Ⅱ, MMP -13, LC-3, glycogenin-1, caspase-3 and make gray analysis. Results: Compared with sham group, the level of collagen Ⅱ decreased, MMP-13 increased, LC-3 decreased, glycogenin-1 increased and caspase-3 increased in experimental group. The apoptosis rate of chondrocytes in experimentaal group [ (17.3±4.4) %] at 72h was higher than that in control group [ (5.6±2.1) %](t=10.732, P<0.001) .WB bands gray statistical results show that the level of collagen Ⅱ in chondrocytes of experimental group (0.43±0.21) was lower than that of control group (0.71±0.26) (t=2.409, P=0.043) , the level of MMP-13 in chondrocytes of experimental group (0.73±0.31) was higher than that of control group (0.24±0.10) (t=3.364, P=0.010) , the level of LC-3 in chondrocytes of experimental group (0.09±0.04) was lower than that of control group (0.39±0.18) (t=3.638, P=0.007) , the level of glycogenin-1 in chondrocytes of experimental group (0.68±0.30) was higher than that of control group (0.29±0.17) (t=2.529, P=0.035) , the level of caspase-3 in chondrocytes of experimental group (0.19±0.08) was higher than that of control group (0.05±0.02) (t=3.796, P=0.005) . Conclusions: The level of autophagy of condylar chondrocytes in temporomandibular joint of rats decreased, glycogen accumulation increased, the rate of chondrocyte apoptosis increased, and the number of chondrocytes decreased, resulting in degeneration of condylar cartilage tissue.

[Research progress on current situation and preventive measures for the narcotic and psychotropic drugs abuse in the elderly].

Both narcotic and psychotropic drugs are more often used to alleviate related multiple physical or mental health problems, but these drugs are very easily addicted to. With the aging of population, abuse of narcotic drugs and psychotropic drugs among the elderly are called for more attention. In this paper, harms caused by the abuse of anesthetic and psychotropic drugs, current situation and causes related to the abuse of anesthetic and psychotropic drugs as well as risk factors and preventive measures regarding the abuse of anesthetic and psychotropic drugs, among the elderly, are reviewed.

[Experimental study on the improvement of Ilizarov transverse tibial bone transport and microcirculation reconstruction technique].

Objective: To observe the effect of traditional and modified Ilizarov transverse tibial bone transport on microvascular regeneration of lower limbs in dogs. Methods: After general anesthesia on 10 experimental dogs, traditional and modified transverse tibial bone transport were performed on both tibias respectively. The control group was treated with the traditional method (the periosteum and bone flap were completely isolated), while the experimental group was treated with the modified method (the fibular periosteum of the open window bone flap was retained). All the external fixators were pulled outwards at a speed of 1 mm every day from 5 days after operation;after one week, the external fixators were moved back every 3 days for one week. The situation of wounds and activity of lower limbs were observed. Simultaneously, the angiogenesis was observed by digital subtraction angiography (DSA) through femoral artery at different stages, and the density of vascular endothelial cells measured by local tissue sections. The data before and after the operation were compared with paired t test. Results: The operation was successfully completed in 10 experimental dogs, and all wounds healed about 1 week after the operation. There was no significant abnormality in lower limb movements in all dogs. Peripheral blood vessel area in middle leg of lower limb in 4 weeks and 8 weeks after operation was (5.9±0.4) mm(2) and (6.9±0.6) mm(2) in control group and it was (6.2±0.6) mm(2) and (8.0±0.6) mm(2) in experimental group, respectively; all were significantly improved than those before the operation ((5.0±0.4) mm(2), (4.9±0.4) mm(2), respectively) (F=446.457, 829.192, both P<0.05). There was no significant differences in vessel area between the two groups at the 4th week after operation (t=1.216, P=0.240), but there was significant difference at the 8th week after operation between the two groups (t=4.423, P=0.000). The percentage of vascular endothelial cells in stained cells under endoscopy was 4.42%±0.28% and 5.63%±0.53% in the control group at the 8th week; and in the experimental group, it was 5.35%±0.26% and 7.18%±0.25%, respectively;all were significantly elevated than those before the operation; and there were significant differences between the two groups (t=7.35, 8.30, both P<0.05). Conclusion: Transverse tibial bone transport and microvascular network regeneration technology can reconstruct the microvasculature below the calf of dogs; the method of window-opening osteotomy is improved to "door" type window-opening, it can retain the lateral periosteum of tibial crest and regenerate the microvasculature network more abundantly.

[Effect of low expression of PTHrp receptor on proliferation of tibial growth plate chondrocytes in chronic renal insufficiency young rats].

Objective: To observe the effect of parathyroid hormone-related protein (PTHrp) receptor on the proliferation of tibial growth plate chondrocytes in chronic renal insufficiency (CRI) young rats. Methods: Two-week-old male SD rats were randomly divided into two groups: (1) Sham group (n=6), only left ureter was exposed; (2) CRI group(n=6), left ureter was ligated to induce chronic renal insufficiency. Rats were sacrificed 2 weeks after operation and the blood concentration of PTHrp was detected by intracardiac blood sampling. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. The level of PTHrp receptor in chondrocytes was observed by immunohistochemistry and quantitative analysis was completed by Western blot. The proliferation rate of chondrocytes from two groups at 24 h was detected by using 5-ethynyl-2'-deoxyuridine (EDU) technique. Three types of PTHrp receptor mRNA plasmids (overexpressed, empty vector and knockdown) were used to treat the chondrocytes from CRI group. The mRNA and protein levels of PTHrp receptor were detected after 24 h and 48 h intervention, respectively. The chondrocyte proliferation rate at 24 h was detected by EDU. Results: Blood concentration of PTHrp in CRI group was higher than that in Sham group [(1.36±0.42) ng/L vs (0.77±0.21) ng/L, t=3.913, P=0.001]. The results of Western blot showed that the level of PTHrp receptor in growth plate chondrocytes from CRI group decreased (0.15±0.07 vs 0.41±0.13, t=5.569, P<0.001). Chondrocyte proliferation rate of CRI group was lower than that in Sham group at 24 h [(11.3±3.1)% vs (24.6±5.7)%, t=6.482, P<0.001]. The mRNA and protein levels of PTHrp receptor increased in chondrocytes of CRI group after intervention with overexpressed plasmid. The chondrocyte proliferation rate increased at 24 h. On the contrary, the mRNA and protein levels of PTHrp receptor decreased afer intervention with knockdown plasmid, and the chondrocyte proliferation rate also decreased [overexpression: (22.8±6.5)%, empty carrier: (10.2±4.3)%, knockdown: (5.6±2.1)%, F=29.840, P<0.001]. Conclusion: Increased PTHrp concentration in the blood of CRI young rats leads to decreased PTHrp receptors in growth plate chondrocytes, which results in decreasing PTHrp activity and proliferation rate of chondrocyte.

A metabolomics approach to characterize raw, pasteurized, and ultra-high temperature milk using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry and multivariate data analysis.

We developed a metabolomics workflow using ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry to determine the effect of thermal treatment on milk composition and metabolites based on multivariate data analysis. We analyzed raw, pasteurized, and UHT milk samples. The samples were first centrifuged to remove the fat layer and mixed with methanol to precipitate proteins. Subsequently, the supernatant was analyzed by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry in electrospray negative mode. Mass spectral data were acquired in MSE mode, a technique whereby both precursor and fragment mass spectral are simultaneously acquired by alternating between low and high collision energy (CE) during a single analytical run, to enable metabolite identification. Based on multivariate data analysis, these markers were significantly affected by thermal treatment. Among the 8 potential markers, we identified 7 oxylipids (9-hydroxydecanoic acid, 12-hydroxydodecanoic acid, 2-hydroxymyristic acid, 3-hydroxytetradecanoic acid, 5-hydroxyeicosatetraenoic acid, 3-hydroxyhexadecanoic acid, and 10-hydroxyoctadecanoic acid) and 1 phospholipid (LysoPE, hexadecanoyl-lysophosphatidylethanolamine). The oxylipids seemed to be adequate for distinguishing UHT milk from raw and pasteurized milk. The structures of the 8 potential markers were identified and characterized using informatics software. Our metabolomics workflow provides a fast approach for the identification of various types of milk.

Genome-wide DNA methylation patterns in coronary heart disease.

BACKGROUND: To better understand the molecular mechanisms of atherosclerosis, we conducted a comparative analysis of DNA methylation patterns in right coronary arteries in the area of advanced atherosclerotic plaques (CAP), great saphenous vein (GSV), and internal mammary artery (IMA) of patients affected by coronary heart disease. METHODS: DNA methylation data (accession number E­GEOD-62867) were divided into three paired groups: CAP vs. IMA, CAP vs. GSV, and IMA vs. GSV. Differentially methylated genes (DMGs) were extracted to analyze the changes in the DMGs in the three different tissues. The gplots package was used for the clustering and heatmap analysis of DMGs. Subsequently, DMG-related pathways were identified using DAVID (Database for Annotation, Visualization and Integrated Discovery) and transcription factors (TFs) were predicted. RESULTS: Based on the filtering criterion of p < 0.05, and a mean beta value difference of ≥0.2, there were 252, 373, and 259 DMGs, respectively, in the CAP vs. IMA, CAP vs. GSV, and IMA vs. GSV groups. Interestingly, the S100A10 gene was hypomethylated in CAP compared with IMA and GSV. Clustering and heatmap analyses suggested that DMGs were segregated into two distinct clusters. Hypermethylated genes in CAP as compared with GSV were only involved in the pathway of fat digestion and absorption, while hypomethylated genes in CAP compared with GSV mainly participated in immune response-associated pathways (cytokine-cytokine receptor interaction, MAPK signaling pathway). CONCLUSION: The DNA methylation differences in vascular tissues of patients with coronary artery disease may provide new insights into the mechanisms underlying the development of atherosclerosis. The functions identified here-cytokine-cytokine receptor interaction, MAPK signaling pathway, DMG (S100A10), and TF (NF-kB)-may serve as potential targets in the treatment of atherosclerosis.

Trends in drug-resistant tuberculosis after the implementation of the DOTS strategy in Shenzhen, China, 2000-2013.

SETTING: The DOTS strategy has been regarded as the most cost-effective way to stop the spread of tuberculosis (TB) since its launch by the World Health Organization. OBJECTIVE: To estimate the effects of DOTS by tracking long-term trends in multidrug-resistant TB (MDR-TB). DESIGN: A retrospective cohort study was conducted from 2000 to 2013 to analyse trends in resistance to anti-tuberculosis drugs and the effect of DOTS-based treatment in Shenzhen, China, using the χ2 test. RESULTS: An overall MDR-TB rate of 4.2% was observed between 2000 and 2013, with an annual reduction of 0.16%. From 2000 to 2013, trends in resistance to isoniazid (INH), rifampicin (RMP) and MDR-TB declined significantly in new TB patients (P < 0.01), but not in retreatment cases. Sputum smear conversion rates after 2 months of treatment decreased significantly, in particular after 2007, in new and retreatment cases. CONCLUSION: INH and RMP resistance and MDR-TB rates declined significantly, suggesting that DOTS-based programmes were successful in reducing drug resistance in new cases but not in retreatment cases. The decreasing sputum smear conversion rates may have been due to an increase in the number of migrants. These two findings suggest that TB is unlikely to be completely eliminated by 2050 in Shenzhen.

CCL2−2518 (A/G) polymorphisms and tuberculosis susceptibility: a meta-analysis.

BACKGROUND: It has been found that the -2518 C-C motif ligand (CCL)-2 promoter variant increases the risk of developing active tuberculosis (TB). OBJECTIVE: To study the association between -2518 variants and susceptibility to TB. DESIGN: We searched Medline, PubMed and the Wan Fang databases for human genetic studies on whether the -2518 CCL2 polymorphism influences the expression of active TB. Articles published from January 1998 to November 2010 were included. A random effects model was conducted in the meta-analysis. RESULTS: The CCL2-2518G allele (OR 1.51, 95%CI 1.11-2.04, P = 0.008) showed significant association with susceptibility to TB. In genotype analysis, the recessive model (CCL2 genotype GG, OR 1.66, 95%CI 1.19-2.33, P = 0.003) was slightly superior to the dominant model (G carrier genotypes OR 1.53, 95%CI 1.07-2.17, P = 0.018). These observations were prominent among Asians and Latin-Americans of Hispanic ancestry, but not in Africans from Ghana and South Africa. The presence of epistatic genes in one population but not in the other, environmental differences and pathogen virulence may account for this. CONCLUSION: The CCL2-2518G allele increases the risk of developing TB in Asians and Hispanics.

Impact of new migrant populations on the spatial distribution of tuberculosis in Beijing.

SETTING AND OBJECTIVES: The tuberculosis (TB) case notification rate in Beijing has been increasing since 2000. Migration was speculated to play an important role in promoting the growth of the epidemic. The identification of spatial clusters of TB can be a key indicator for targeting limited public health resources. METHODS: Spatial modelling was applied to the 220 towns of Beijing and summarised for the 18 districts, which were combined into four functional areas in 2005 and 2006. Population density was combined with the numbers of TB cases, and TB incidence data was used to identify high rate clusters. A negative binomial regression model was used to confirm the association between TB and migration status in Beijing. RESULTS: There were 4584 TB cases among permanent residents and 2838 among migrants. TB cases and population were most densely grouped in four central districts. High-rate TB clusters in both permanent residents and migrants were detected in the 'New Districts for Urban Development' and Chaoyang District in 2005 and 2006. Migration and the population growth rate of new migrants are contributing to the TB increase in Beijing. CONCLUSIONS: The increasing migrant population has had a drastic influence on the spatial distribution of TB in Beijing. Spatial analysis could provide additional information in addition to common incidence plots.

Heteroclitin H, a new lignan from Kadsura heteroclita.

A new dibenzocyclooctadiene lignan, heteroclitin H (1), was isolated together with seven known compounds, heteroclitin D (2), interiorin B (3), interiorin (4), neokasuranin (5), interiotherin C (6), gomisin J (7) and (+)-anwulignan (8), from the stems of Kadsura heteroclita. Their structures were elucidated by spectroscopic methods. This is the first report of the isolation of compounds 3, 5, 6, 7 and 8 from K. heteroclita.