Impact of preoperative balance training on postoperative functional recovery of patients after total knee arthroplasty: a systematic review and meta-analysis.

OBJECTIVE: Previous studies' results on the impact of preoperative balance training on postoperative functional recovery after total knee arthroplasty (TKA) appeared to be ambiguous. Thus, this systematic review and meta-analysis were performed to investigate the effects of preoperative balance training on walking ability, balance-specific performance, and other functional indicators in elderly patients post-TKA. MATERIALS AND METHODS: Patient data were obtained from databases including PubMed, Physiotherapy Evidence Database (PEDro), CINAHL, SPORTDiscus, and Scopus. The inclusion criteria followed the Population-Intervention-Comparison-Outcome (PICO) principle. The assessment process involved meticulous screening, judicious data extraction, and rigorous evaluation of trial method quality, conducted by two independent researchers. Based on standardized mean differences and 95% confidence intervals, meta-analysis was performed employing a random-effects model or fixed-effects model. RESULTS: Preoperative balance training appears to be a potentially effective intervention for enhancing the knee osteoarthritis (KOA) patients' knee joint function (RR = 1.16, 95% CI: -2.58, 4.91), isometric knee flexion (RR = 2.49, 95% CI: -2.53, 7.50), knee extension (RR = -0.13, 95% CI: -0.45, 0.18), knee society score (KSS) (RR = 2.18, 95% CI: -1.51, 5.88), stair test (RR = -0.73, 95% CI: -1.84, 0.37), and timed up and go (RR = -1.18, 95% CI: -1.60, -0.76). CONCLUSIONS: Compared to interventions with less emphasis on balance training, rehabilitation programs highly emphasizing balance training significantly enhance the walking ability, balance specificity, and functional indicators of elderly patients post-TKA. This includes rehabilitation programs for senior TKA patients, with a focus on activities meant to improve the sensory system, balance in particular.

Application of personalized spacers made with 3D-printed mold in the two-stage procedure for periarticular infection.

BACKGROUND: Cement spacers treat periarticular infection after bone tumor resection in patients with bone defects. Complications such as poor joint function, poor soft tissue reconstruction, and poor postoperative daily living ability are present. We present a case of periarticular infection treated successfully after distal femoral osteosarcoma surgery with a personalized spacer made with a 3D-printed mold. CASE REPORT: A two-stage procedure was performed on an 18-year-old patient with high-grade conventional osteosarcoma of the left distal femur. After two biopsies, the boy developed a periarticular infection of the affected limb during neoadjuvant chemotherapy. We had a microbiologically confirmed methicillin-resistant Staphylococcus aureus (MRSA) infection. Because of the infection risk associated with primary joint replacement, a two-stage procedure was planned. In the first stage of surgery, we prepared a personalized spacer using a 3D-printed mold, antibiotic-loaded polymethylmethacrylate (PMMA), and an intramedullary needle. This spacer restored the function of the knee joint and the daily activities of the affected limb, and the infection was effectively eradicated. This spacer was firmly fixed two years after the surgery, and there were no surgical or spacer-related complications. The patient underwent a second stage of surgery to replace a permanent metal mega-prosthesis, and the knee joint functions returned to near normal. CONCLUSIONS: This case report describes limb-salvage surgery following distal femoral resection for periarticular infection. The personalized spacers prepared by a 3D-printed mold can be used in periarticular infection after long bone resection, mega-prosthetic infection, or limb-salvage surgery for temporary joints in small children.

[Advances of hydrogel in early protection of exposed intra-abdominal organs after open abdomen].

The protection of open abdomen (OA) wound is a significant subject in the field of trauma surgery. The key technical challenge in the early stage of OA wound management involves promoting granulation tissue filling between intestinal segments, reducing intestinal wall abrasion, and preventing the development of enteroatmospheric fistulas (EAF). Hydrogels, characterized by their high water content and exceptional biocompatibility, serve as extracellular matrix-mimicking materials, and are extensively employed in various medical and healthcare applications. In this review, we discuss the application of hydrogel developed by natural biomaterials in OA wounds protection, taking into consideration the unique pathophysiological characteristics of the OA wounds. This review aims to provide valuable insights for the development of hydrogel materials for early-stage OA wound protection in future research.

The axial and sagittal CT values of the 7th thoracic vertebrae in screening for osteoporosis and osteopenia.

AIM: To evaluate the difference in computed tomography (CT) attenuation value of different planes of the 7th thoracic vertebra and investigate the efficacy of axial and sagittal vertebral CT measurements in predicting osteoporosis. MATERIALS AND METHODS: Patients who underwent routine chest CT and dual-energy X-ray absorptiometry (DXA) within 1 month were included in this retrospective study. The CT attenuation values of different planes were compared. Logistic regression and receiver operating characteristic (ROC) were used to analyse the difference of each plane in the diagnosis of osteoporosis. RESULTS: The study included 1,338 patients (mean age of 61.9±11.9; 54% female). The CT attenuation values decreased successively in the normal group, osteopenia group, and osteoporosis group. The paired t-test results showed that the mid-axial measurements were greater than mid-sagittal measurements, with a mean difference of 9 HU, the difference was statistically significant (p<0.001, 95% confidence interval [CI] = 7.8-10.1). For each one-unit reduction in mid-sagittal CT attenuation value, the risk of osteopenia or osteoporosis increased by 3.6%. To distinguish osteoporosis from non-osteoporosis (osteopenia + normal), the sensitivity was 90% and the specificity was 52.4% at the mid-sagittal threshold of 113.7 HU. CONCLUSIONS: The CT attenuation values of mid-sagittal plane have higher diagnostic efficacy than axial planes in predicting osteoporosis. For patients with a sagittal CT attenuation value of <113.7 HU in the T7, further DXA examination is warranted.

[Molluscicidal activity of the secondary metabolites from Streptomyces nigrogriseolus XD 2-7 against Oncomelania hupensis and its preliminary mechanisms of molluscicidal actions].

OBJECTIVE: To evaluate the storage stability of metabolites from actinomycetes Streptomyces nigrogriseolus XD 2-7 and the mollcuscicidal activity against Oncomelania hupensis in the laboratory, and to preliminarily explore the mechanisms of the molluscicidal activity. METHODS: The fermentation supernatant of S. nigrogriseolus XD 2-7 was prepared and stored at -20, 4 °C and 28 °C without light for 10 d; then, the molluscicidal effect was tested against O. hupensis following immersion for 72 h. The fermentation supernatant was boiled in a 100 °C water bath for 30 min and recovered to room temperature, and then the molluscicidal effect was tested against O. hupensis following immersion for 72 h. The pH values of the fermentation supernatant were adjusted to 4.0, 6.0 and 9.0 with concentrated hydrochloric acid and sodium hydroxide, and the fermentation supernatant was stilled at room temperature for 12 h, with its pH adjusted to 7.0; then, the molluscicidal effect was tested against O. hupensis following immersion for 72 h. The fermentation product of S. nigrogriseolus XD 2-7was isolated and purified four times with macroporous resin, silica gel and octadecylsilane bonded silica gel. The final products were prepared into solutions at concentrations of 10.00, 5.00, 2.50, 1.25 mg/L and 0.63 mg/L, and the molluscicidal effect of the final productswas tested against O. hupensis following immersion for 72 h, while dechlorination water served as blank controls, and 0.10 mg/L niclosamide served as positive control. The adenosine triphosphate (ATP) and adenosine diphosphate (ADP) levels were measured in in O. hupensis soft tissues using high performance liquid chromatography (HPLC) following exposure to the final purified fermentation products of S. nigrogriseolus XD 2-7. RESULTS: After the fermentation supernatant of S. nigrogriseolus XD 2-7 was placed at -20, 4 °C and 28 °C without light for 10 d, immersion in the stock solution and solutions at 10- and 50-fold dilutions for 72 h resulted in a 100% (30/30) O. hupensis mortality. Following boiling at 100 °C for 30 min, immersion in the stock solution and solutions at 10- and 50-fold dilutions for 72 h resulted in a 100.00% (30/30) O. hupensis mortality. Following storage at pH values of 4.0 and 6.0 for 12 h, immersion in the fermentation supernatant of S. nigrogriseolus XD 2-7 for 72 h resulted in a 100.00% (30/30) O. hupensis mortality, and following storage at a pH value of 9.0 for 12 h, immersion in the fermentation supernatant of S. nigrogriseolus XD 2-7 for 72 h resulted in a 33.33% (10/30) O. hupensis mortality (χ2 = 30.000, P < 0.05). The minimum concentration of the final purified fermentation products of S. nigrogriseolus XD 2-7 was 1.25 mg/L for achieving a 100% (30/30) O. hupensis mortality. The ATP level was significantly lower in O. hupensis soft tissues exposed to 0.10 mg/L and 1.00 mg/L of the final purified fermentation products of S. nigrogriseolus XD 2-7 than in controls (F = 7.274, P < 0.05), while no significant difference was detected in the ADP level between the treatment group and controls (F = 2.485, P > 0.05). CONCLUSIONS: The active mollcuscicidal ingredients of the S. nigrogriseolus XD 2-7 metabolites are maintained stably at -20, 4 °C and 28 °C for 10 d, and are heat and acid resistant but not alkali resistant. The metabolites from S. nigrogriseolus XD 2-7 may cause energy metabolism disorders in O. hupensis, leading to O. hupensis death.

[Activity of aromatic pyrrole-based compounds against of Schistosoma japonicum cercariae and acute toxicity to fish].

OBJECTIVE: To test the activity of aromatic pyrrole-based compounds against cercariae of Schistosoma japonicum and test their acute toxicity to fish. METHODS: A series of aromatic pyrrole-based compounds were synthesized using 4-benzyl-5-(trifluoromethyl)-1H-pyrrole-3-nitrile as the lead compound. The synthesized compounds were prepared into solutions at concentrations of 10.00, 1.00, 0.10, 0.01 mg/L, and the activity of these solutions against S. japonicum cercariae was tested in 30 min, while 0.10 mg/L and 0.01 mg/L niclosamide solutions served as a positive control and dechlorinated water with 1% dimethyl sulfoxide (DMSO) was used as a negative control, with 10 to 30 cercariae of S. japonicum in each group. In addition, the compounds were prepared into solutions at concentrations of 0.50, 0.25, 0.12, 0.06, 0.03 mg/L, and their toxicity to zebrafish was tested in 72 h, while 0.15 mg/L and 0.30 mg/L niclosamide solutions served as a positive control and dechlorinated water with 1% DMSO was used as a negative control, with 10 zebrafishes in each group. RESULTS: A total of 7 aromatic pyrrole-based compounds were successfully synthesized. Treatment with compounds 102, 104 and 106 at a concentration of 0.01 mg/L for 30 min killed all S. japonicum cercariae, and compounds 105 and 107 showed no activity against cercariae. No death of cercariae was found in the blank control group, while treatment with 0.10 mg/L niclosamide for 10 min caused a 100% mortality rate of S. japonicum cercariae and 0.01 mg/L niclosamide failed to kill S. japonicum cercariae. No zebrafish death was found 72 h post-treatment with compounds 101, 104 and 105 at a concentration of 0.03 mg/L, and exposure to compounds 102, 103 and 106 at a concentration of 0.03 mg/L for 12 h resulted in a 100% mortality rate of zebrafish. No zebrafish death occurred 72 h post-treatment with 0.50 mg/L Compound 104, and no zebrafish death was found in the blank control group, while treatment with 0.30 mg/L niclosamide for 24 h resulted in a 100% mortality rate of zebrafish. CONCLUSIONS: Compound 104 achieves a 100% mortality rate against S. japonicum cercariae at a concentration of 0.01 mg/L for 30 min, and causes no death of zebrafish at a concentration of 0.50 mg/L for 72 h, which may serve as a cercaricide candidate.

[Effects of praziquantel isomers on the proliferation and activation of the LX-2 human hepatic stellate cell line].

OBJECTIVE: To compare the effects of levo-praziquantel (L-PZQ) and dextro-praziquantel (D-PZQ) on the proliferation and activation of the human hepatic stellate cell line LX-2 in vitro. METHODS: LX-2 cells were stimulated with transforming growth factor-ß (TGF-ß). LX-2 cell proliferation was measured using the CCK-8 assay after 24 h stimulation with 0 to 50 µg/mL concentrations of praziquantel, and the gene and protein expression of type Ⅰ collagen (collagen Ⅰ), type Ⅲ collagen (collagen Ⅲ) and α-smooth muscle actin (α-SMA) was quantified in LX-2 cells using quantitative real-time PCR (qPCR) and Western blotting assays 24 h and 48 h following stimulation with 15 µg/mL praziquantel to detect LX-2 cell activation. RESULTS: There were significant differences in the survival rate of LX-2 cells between L-PZQ and D-PZQ treatments at all concentrations (F = 6.119 and 79.180, both P values < 0.05). Either L-PZQ or D-PZQ at a concentration of < 30 µg/mL showed no remarkableeffectsonthe LX-2 cell proliferation (both P values > 0.05), and L-PZQ at a concentration of > 50 µg/mL and D-PZQ at a concentration of > 40 µg/mL inhibited the LX-2 cell proliferation (both P values < 0.05), while D-PZQ at concentrations of 40 µg/mL and 50 µg/mL showed greater inhibition on LX-2 cell proliferation than L-PZQ (t = 3.419 and 8.776, both P values < 0.05). There were significant differences in the collagen Ⅰ, collagen Ⅲ and α-SMA expression in LX-2 cells at both transcriptional (F = 21.55, 79.99 and 46.70, all P values < 0.05) and translational levels (F = 20.12, 30.29 and 32.93, all P values < 0.05) among the blank control group, TGF-ß stimulation group, L-PZQ treatment group and D-PZQ treatment group. L-PZQ treatment resulted in remarkable inhibition on collagen Ⅲ and α-SMA gene expression in LX-2 cells (both P values < 0.05); however, the treatment showed no remarkable inhibition collagen Ⅰ gene expression or collagen Ⅰ, collagen Ⅲ or α-SMA protein expression in LX-2 cells (all P values > 0.05). In addition, D-PZQ treatment resulted in significant inhibition on collagen Ⅰ, collagen Ⅲ and α-SMA expression in LX-2 cells at both translational and transcriptional levels (all P values < 0.05), and D-PZQ showed higher inhibition on collagen Ⅰ, collagen Ⅲ and α-SMA gene expression in LX-2 cells than L-PZQ (all P values < 0.05). CONCLUSIONS: Both L-PZQ and D-PZQ inhibit the proliferation and activation of LX-2 cells, and D-PZQ shows a higher inhibitory activity than L-PZQ.

[Studies on resistance of Schistosoma to praziquantel XVIII Sensitivity to praziquantel in filial generations of praziquantel-resistant and -sensitive Schistosoma japonicum mixed infections].

OBJECTIVE: To investigate the sensitivity of adult worms of filial generations from praziquantel-resistant and -sensitive Schistosoma japonicum mixed infections to praziquantel. METHODS: Mice were infected with the cercariae of an experimentally generated praziquantel-resistant S. japonicum isolate [median effective dose (ED50) = 277.4 mg/kg] and a laboratory-maintained praziquantel-sensitive S. japonicum isolate (ED50 = 99.6 mg/kg) at a mixture ratio of 1:1 and 2:1, which was maintained in the laboratory via the mouse-snail cycle for 8 generations. Then, mice were infected with the cercariae of the 8th filial-generation parasite, and grouped 35 days post-infection. Mice in the 5 treatment groups were given praziquantel treatment by gavage at a single oral dose of 37.5, 75, 150, 300 mg/kg and 600 mg/kg, while animals in the control group was administered orally with 2.5% cremophor EL. All mice were sacrificed 14 days post-treatment and adult worms were collected by perfusion of the portal vein. The worm burden reductions and praziquantel ED50 values were calculated. The praziquantel-resistant S. japonicum isolate generated from experimental induction with 12 rounds of praziquantel treatment with sub-curative doses was maintained in the laboratory via the mouse-snail cycle, and mice were infected with the cercariae of the 8th filial-generation parasite. The praziquantel ED50 value against the 8th filial-generation adults was measured. RESULTS: After mice were infected with the mixture of cercariae of PZQ-resistant and -sensitive S. japonicum isolates at a ratio of 1:1, the praziquantel ED50 was 135.2 mg/kg against the adults of the 8th filial-generation parasite. After mice were infected with the mixture of cercariae of PZQ-resistant and -sensitive S. japonicum isolates at a ratio of 2:1, the praziquantel ED50 was 129.2 mg/kg against the adults of the 8th filial-generation parasite. In addition, the praziquantel ED50 was 208.4 mg/kg against the adults of the 8th filial-generation S. japonicum without the selection pressure of praziquantel. CONCLUSIONS: Compared with the experimentally induced praziquantel-resistant S. japonicum isolate, the adult worms of the filial-generation S. japonicum show a reduced sensitivity to praziquantel in the same host following infection with the mixture of cercariae of praziquantel-resistant and -sensitive S. japonicum isolates. The adult worms of the filial generation of the praziquantel-resistant S. japonicum isolate without the selection pressure of praziquantel may still maintain the resistance to praziquantel.

[Study on the factors affecting the degradation of niclosamide in the soil].

OBJECTIVE: To investigate the factors affecting the degradation of niclosamide in the soil, so as to provide the evidence for the assessment of the environmental safety in the field snail control with niclosamide. METHODS: A high performance liquid chromatography was established for the determination of niclosamide in the field. Then, the degradation of niclosamide was investigated in soils with different moistures (10%, 30%, 50%, 70% and 90%), temperatures [(15 ± 1), (25 ± 1), (35 ± 1) °C], initial concentrations (1, 5, 10 mg/kg) and in sterilized and non-sterilized soils. In addition, the degradation of niclosamide was fitted with the first-order kinetics equation, and the degradation half-life was calculated. RESULTS: The niclosamide residues gradually decreased over time in soils with different moistures, and a higher rate of degradation was seen in soils with a higher moisture. The degradation half-life of niclosamide reduced from 4.258 d in the soil with a 10% moisture to 2.412 d in the soil with a 90% moisture. The niclosamide residues gradually decreased over time in soils with different temperatures, and a higher rate of degradation was seen in soils with a higher temperature. The degradation half-life of niclosamide reduced from 4.398 d in the soil with a temperature of (15 ± 1) °C to 2.828 d in the soil with a temperature of (35 ± 1) °C. The degradation half-lives of niclosamide were 3.212, 3.333 d and 3.448 d in soils containing niclosamide at initial concentrations of 1, 5 mg/kg and 10 mg/kg, and > 30 d and 3.273 d in sterilized and non-sterilized soils. Multiple linear regression analysis revealed that soil microorganisms (P = 0.010), moisture (P = 0.000) and temperature (P = 0.002) affected the half-life of niclosamide degradation. CONCLUSIONS: The degradation of niclosamide in soils fits the first-order kinetics equation, and presence of microorganisms, a high temperature and high moisture may accelerate the degradation of niclosamide in the soil.

Measurement of Scars Using Structured-Light 3D Scanning in Forensic Practice.

ABSTRACT: Objective To validate the accuracy and reliability of structured-light three-dimensional （SL-3D） scanning in measuring the length and area of the regular and irregular scars on body surface and discuss its value in forensic practice. Methods The lengths of 30 cases of simulated linear scars and 50 cases of linear scars after injury were measured using soft ruler, vernier caliper + thin line method, and SL-3D scanning. The areas of 35 cases of simulated patchy scars and 15 cases of patchy scars after injury were measured using length × width, film tracing with coordinate paper method, pixel method, and SL-3D scanning, and then statistically analyzed. Results The differences between the length of the simulated linear scars measured by SL-3D scanning and standard length had no statistical significance. When simulated patchy scars and patchy scars after injury were measured with high surface curvature and large irregular areas, the differences between the results of SL-3D scanning measurement and the standard area had no statistical significance. When the length of 50 cases of linear scars after injury were measured using SL-3D scanning, the correlation coefficient between the measurement results of two different investigators was 0.998, and the correlation coefficient between the two measurement results by the same investigator was 1.000. The correlation coefficient between the results of SL-3D scanning and that of vernier caliper + thin line method was 0.996. Conclusion The three-dimensional information of the scars on the body surface can be acquired using SL-3D scanning. The measurement of the length and area of the scars is not influenced by the location of scars, curvature of surface, and human factors. The measurement results are accurate, reliable and has unique advantages.

Recombinant lactobacillin PlnK adjusts the gut microbiome distribution in broilers.

1. The heterologous expression and biological function of the Lactobacillus bacteriocin plantaricin K (PlnK) remain largely unknown. 2. In this study, PlnK was efficiently expressed in competent E. coli BL21 (used in transformation and protein expression) after 12 h, at 37°C and in 0.4 mmol/l isopropyl ß- d-1-thiogalactopyranoside (IPTG). 3. The inhibitory bacterial spectrum of recombinant PlnK was investigated and indicated that levels of PlnK above 0.10 mg/ml produced an obvious inhibitory effect on gram-positive bacteria and gram-negative bacteria in vitro. 4. The effects of PlnK on intestinal immune function and the gut microbiome distribution in broilers were studied. The results revealed that, after consuming 2.50 × 10-3 mg/ml of PlnK in water for one week, at the phylum level, the abundance of Firmicutes was increased and the abundance of Bacleroidetes was decreased. At the family level, the abundance of Lachnospiraceae, Ruminococcaceae and Streptococcaceae were significantly improved, but the abundance of Bacteroidaceae was reduced. At the genus level, the abundances of Lachnoclostridium, Streptococcus and Ruminococcaceae-UCG-013, were significantly up-regulated, and the abundance of Bacteroides was down-regulated. 5. After oral liquid intake of PlnK for one week, levels of secretory immunoglobulin A (sIgA) in the duodenal mucus were not significantly increased, but the mRNA levels of TLR3, MDA5, IFN-α, IFN-ß, IFITM3 and IFITM10 in the duodenum were significantly reduced. 6. This study demonstrated that the recombinant PlnK could adjust the intestinal microbiome distribution and downregulate the IFN pathway.

Association between caregivers' anxiety and depression symptoms and feeding difficulties of preschool children: A cross-sectional study in rural China.

PURPOSE: To investigate the prevalence of feeding difficulties in preschool children and explore the association between caregivers' anxiety and depression symptoms and preschool children's feeding difficulties. METHODS: This cross-sectional study was conducted between June 2017 and January 2018 in rural areas of Anhui province, China. A total of 2231 preschool children and their caregivers were interviewed. Feeding difficulties of preschool children were reported by caregivers using the adapted Identification and Management of Feeding Difficulties (IMFeD) tool. Anxiety and depression symptoms of caregivers were evaluated via the Self-Rating Anxiety Scale (SAS) and the Self-Rating Depression Scale (SDS). RESULTS: In total, 54.1% of preschool children were reported to have feeding difficulties by their caregivers. Among all children, when the caregivers had symptoms of anxiety or depression, the children had a higher risk of feeding difficulties. Specifically, for caregivers' anxiety symptoms, the odds ratios (ORs) of feeding difficulties in all children, left-behind children (LBC), and non-LBC were 1.91 (95% confidence interval [CI]: 1.42-2.57), 2.04 (95% CI: 1.34-3.09), and 1.86 (95% CI: 1.21-2.87), respectively; for caregivers' depression symptoms, the ORs of feeding difficulties in all children, LBC, and non-LBC were 1.86 (95% CI: 1.46-2.39), 1.76 (95% CI: 1.24-2.51), and 2.08 (95% CI: 1.45-2.97), respectively. In addition, when caregivers who were parents or grandparents had anxiety or depression symptoms, their children had a higher risk of feeding difficulties. Specifically, for parents and grandparents with anxiety symptoms, the ORs of feeding difficulties were 1.84 (95% CI: 1.14-2.98) and 2.17 (95% CI: 1.46-3.22), respectively; for parents and grandparents with depression symptoms, the ORs of feeding difficulties were 2.03 (95% CI: 1.40-2.95) and 1.93 (95% CI: 1.37-2.73), respectively. CONCLUSION: Caregivers' anxiety or depression symptoms are positively associated with feeding difficulties in children.

MicroRNA-579-3p promotes the progression of osteoporosis by inhibiting osteogenic differentiation of mesenchymal stem cells through regulating Sirt1.

OBJECTIVE: To explore whether microRNA-579-3P was involved in the development of osteoporosis, and to investigate the possible molecular mechanisms. PATIENTS AND METHODS: The messenger RNA (mRNA) expression levels of microRNA-579-3P, alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2) and bone sialoprotein (BSP) in serum samples of osteoporosis patients and normal controls were detected by quantitative Real-time polymerase chain reaction (qRT-PCR), respectively. Meanwhile, the expressions of the above genes during osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) were examined as well. To investigate the effect of microRNA-579-3P on osteogenesis, microRNA-579-3P was overexpressed and knocked down in hMSCs. Subsequently, the mRNA and protein expression levels of osteogenesis-related genes, such as ALP, RUNX2 and BSP, were detected by qRT-PCR and Western blot, respectively. In addition, ALP activity and mineralization forming ability were evaluated by ALP staining and alizarin red staining. Bioinformatics predicted that Sirt1 was the target gene of microRNA-579-3P. Subsequent luciferase reporter gene assay was performed to verify the binding relationship of microRNA-579-3P to Sirt1. Meanwhile, qRT-PCR and Western blot were used to detect the changes in the mRNA and protein expression levels of Sirt1, respectively. After overexpression of microRNA-579-3P and Sirt1, qRT-PCR, Western blot, ALP staining and alizarin red staining assays were performed to detect the osteogenic differentiation of hMSCs. RESULTS: The expression of microRNA-579-3P in serum of patients with osteoporosis was significantly higher than that of normal controls. Meanwhile, the expression of microRNA-579-3P decreased gradually during osteogenic differentiation of hMSCs. Overexpression of microRNA-579-3P significantly reduced the expressions of osteogenic related genes, including ALP, RUNX2 and BSP. Besides, ALP activity and mineralized nodule formation ability decreased obviously as well. Luciferase reporter gene assay showed that microRNA-579-3P could bind to Sirt1. After overexpression of microRNA-579-3P, the mRNA and protein expression levels of Sirt1 were significantly reduced, which were reversed after silence of microRNA-579-3P. Simultaneous overexpression of microRNA-579-3P and Sirt1 could reverse the inhibition of osteogenic differentiation of hMSCs caused by overexpression of microRNA-579-3P alone. CONCLUSIONS: MicroRNA-579-3P could inhibit osteogenic differentiation of hMSCs by regulating Sirt1, thereby promoting the development of osteoporosis.

[Effect of liraglutide on glucagon secretion in obese type 2 diabetic patients].

Objective: To investigate the effect of liraglutide on glucagon release in obese type 2 diabetes (T2DM). Methods: A multi-center, prospective, and self-comparison study was conducted in four hospitals in Qingdao. Twenty-four patients with T2DM were selected and treated with liraglutide for 12 weeks. Glucagon levels before and after treatment were detected before and 30 min, 60 min and 120 min after meals. Results: After 12 weeks of treatment, the overall level of glucagon decreased, in which the differences in glucagon levels at 30 min [(220±79) ng/L vs. (203±77) ng/L, P<0.05] and 60 min [(248±119) ng/L vs. (203±82)ng/L, P<0.05] reached significance, respectively, comparing to those before treatment. The area under the curve of glucagon after treatment was significantly lower than that before treatment (438±190 vs. 389±153, P<0.05). In contrast, after treatment, the overall level of C-peptide increased, especially the levels at 30 min [(1.53±1.02) nmol/L vs.(2.03±1.29) nmol/L], 60 min [(1.93±1.19) nmol/L vs. (2.48±1.75) nmol/L] and 120 min [(2.36±1.47) nmol/L vs. (2.96±1.84) nmol/L], all P<0.05. The area under C-peptide curve increased significantly (3.6±2.2 vs. 4.6±2.9, P<0.05). Fasting plasma glucose, postprandial 2 h plasma glucose and glycosylated hemoglobin A1c were all lower than before, and the differences were statistically significant (P<0.05). Waist circumference and body mass index were significantly lower than before (P<0.05). The amount of insulin used for the treatment decreased by approximately 55.1% compared with that before liraglutide, and the difference was statistically significant (P<0.05). Conclusions: Liraglutide inhibits glucagon secretion and lowers blood glucose. It can also reduce body weight, improve islet cell function and reduce insulin use in T2DM.

The identification of multifocal breast cancer-associated long non-coding RNAs.

OBJECTIVE: To identify long non-coding RNAs (lncRNAs), which may be associated with multifocal and multicentric breast cancer (MMBC) by analyzing the differential expression of non-coding RNA in MMBC and unifocal breast cancers (UBC). PATIENTS AND METHODS: 156 cases of invasive MMBC patients (136 patients with 2 focuses and 20 patients with 3 focuses) and 130 cases of UBC were collected from January 2005 to December 2015 in Yantai Yuhuangding Hospital. The differentially expressed lncRNAs in MMBC and UBC were screened by gene chip. RT-PCR was used to verify the differentially expressed lncRNAs. RESULTS: Significantly different expression was found in 1080 lncRNAs between MMBC and UBC (FC > 4, p < 0.05), among which, 458 were upregulated and 622 were down-regulated. In homologous lncRNAs, the expression levels of four lncRNAs including C19orf33, C3orf52, C15orf48 and C4orf19 in MMBC tissue were significantly different than those in UBC tissues. RT-PCR verified that the expression of C19orf33, C3orf52 and C15orf48 in MMBC tissue was significantly up-regulated and the expression of C4orf19 was significantly down-regulated, which was consistent with that of gene chip. CONCLUSIONS: C19orf33, C3orf52, C15orf48 and C4orf19 may have important functions in MMBC, and may be used as markers for MMBC.

Continued EGFR-TKIs treatment promotes the survival of elderly patients with acquired resistance to EGFR-TKIs therapy.

OBJECTIVE: Continued EGFR-TKIs treatment is still controversial for NSCLC patients with activating EGFR mutations, who acquire resistance to the drug. Of these patients, elderly ones were worth to be investigated to further examine efficacy of continued EGFR-TKIs treatment. PATIENTS AND METHODS: A total of 232 NSCLC patients (≥ 70-year-old) were recruited from the Chinese People's Liberation Army General Hospital between January 1, 2009, and July 31, 2014. And 44 patients were qualified for further retrospectively investigated, which were divided into dramatic and non-dramatic progression groups based on the characteristics of progression during first-line EGFR-TKIs treatment. And they were also divided into two groups: continued EGFR-TKIs group and discontinued EGFR-TKIs group. Subsequently, progression-free survival (PFS), post-progression survival (PPS), and overall survival (OS) of these groups were investigated by multivariate analysis. RESULTS: Median OS (28.9 months vs. 23.2 months, p = 0.46) and median PPS (16.9 months vs. 4.4 months, p = 0.216) were both not significantly different between continued EGFR-TKIs groups and discontinued ones. However, when focusing on patients with non-dramatic progression, the median OS (29.0 months vs. 23.2 months, p = 0.039) and median PPS (21.3 months vs. 3.9 months, p = 0.001) were significantly longer in the continued EGFR-TKIs patients than discontinued ones. DISCUSSION: Continued EGFR-TKIs beyond PD may be a good option for elderly patients with non-dramatic progression. The characteristic of progression after first-line EGFR-TKIs treatment should be taken into account to determine which part of patients is suitable for continued EGFR-TKIs treatment, especially for the speed of progression. CONCLUSION: Continued EGFR-TKIs treatment promotes the survival of elderly patients with acquired resistance to EGFR-TKIs therapy.

Postnatal development of GABAergic interneurons in the neocortical subplate of mice.

The subplate (SP) plays important roles in developmental and functional events in the neocortex, such as thalamocortical and corticofugal projection, cortical oscillation generation and corticocortical connectivity. Although accumulated evidence indicates that SP interneurons are crucial for SP function, the molecular composition of SP interneurons as well as their developmental profile and distribution remain largely unclear. In this study, we systematically investigated dynamic development of SP thickness and chemical marker expression in SP interneurons in distinct cortical regions during the first postnatal month. We found that, although the relative area of the SP in the cerebral cortex significantly declined with postnatal development, the absolute thickness did not change markedly. We also found that somatostatin (SOM), the ionotropic serotonin receptor 3A (5HT3AR), and parvalbumin (PV) reliably identify three distinct non-overlapping subpopulations of SP interneurons. The SOM group, which represents ~30% of total SP interneurons, expresses neuronal nitric oxide synthase (nNOS) and calbindin (CB) and colocalizes entirely with neuropeptide Y (NPY). The 5HT3AR group, which accounts for ~60% of the total interneuronal population, expresses calretinin (CR) and GABA-A receptor subunit delta (GABAARδ). The PV group accounts for ~10% of total SP interneurons and coexpressed GABAARδ. Moreover, distinct interneuron subtypes show characteristic temporal and spatial distribution in the SP. nNOS(+) interneurons in the SP increase from the anterior motor cortex to posterior visual cortex, while CR(+) and CB(+) interneurons the opposite. Interestedly, the majority of GABAARδ(+) neurons in SP are non-GABAergic neurons in contrast to other cortical layers. These findings clarify and extend our understanding of SP interneurons in the developing cerebral cortex and will underpin further study of SP function.

Soil quality indicators response to application of hydrophilic polymers to a soil from a sulfide mine.

In soils impacted by mining activities a vegetal cover is required to protect the site from the erosive forces of water and wind. The success of this objective depends on plant establishment and canopy closure. Polyacryalate polymers aid the growth of crops and indigenous plants in soils from sulfide mines. Soil characteristics change as a consequence of polymer application, but indicators that pinpoint these changes have not been identified yet. Our objectives were to (1) identify the sensitive indicators of changes in soil quality following polymer application, (2) relate these with assessment based on plant growth and soil cover. A mine soil was left unamended or received a characterized polyacrylate, a polyacrylate removed from diapers, or shredded diapers. Biomass of Spergularia purpurea was measured and proportion of soil cover evaluated. Soil enzymes, microbial activity, and respiration were analyzed. Availability of potentially toxic trace elements was estimated by their concentration in shoots. Factor analysis identified three factors that accounted for 94% of the variation in parameters, and the scores separated the four treatments. The indicators with greatest communality were correlated with plant growth and soil cover. The best soil quality indicators were As and Zn in shoots, protease, ß-glucosidase, and fructose-induced respiration. It seems that the most important indicators to be used to assess the restoration of sulfide mine soils are those related with bioavailability of trace elements and soil enzymatic activities.