Correction: Protein kinase Mζ in medial prefrontal cortex mediates depressive-like behavior and antidepressant response.

Following publication of this article, the authors noted that the bands of Na-K ATPase in Figure S1B were mislabelled when selecting representative blots, and were erroneously duplicated from GluA2. However, the data analysis was still made with the correct immunoblots. The Supplementary Information file has now been updated to include the corrected Figure S1 with the correct western blot bands of Na-K ATPase.The authors would like to apologise for this error and the inconvenience this may have caused. This has been corrected in both the PDF and HTML versions of the article.

[Ultrastructural features of the corneal epithelium with micromolecular compound J2 in the corneal allograft of rats using atomic force microscopy].

Objective: To study the ultrastructural features of the corneal epithelium in the corneal allograft of rats with micromolecular compound J2. Methods: An experimental study. Primarily cultured corneal epithelial cells of rats were used. Mononuclear cells (MNCs) by density gradient centrifugation were assigned into the experimental group [MNCs (2 ml)+corneal epithelial cells+J2], control group [MNCs (2 ml)+corneal epithelial cells] and blank group (corneal epithelial cells). Quantity of CD80 expression was obtained by flow cytometry after coculture. Amplitude and height images were obtained by tapping mode atomic force microscopy (AFM) with a scan rate of 2 Hz and an integral gain of 0.3 to 0.5. Statistical analysis of Ra, Rq, Rvm and Rt was performed using the single-factor analysis of variance, and P value was calculated. Results: There were obvious differences in the ultrastructure measured by AFM among groups. Ra was 86.75±12.60 in the experimental group, 120.23±12.11 in the control group, and 61.94±10.62 in the blank group (F=306.92, P<0.01). Rq was 102.53±9.45, 138.30±10.13, and 91.96±7.25, respectively, in the three groups (F=361.85, P<0.01). Rvm was -42.21±14.22, -67.36±10.89, and -32.18±19.01, respectively (F=72.22, P<0.01). Rt was 437.32±15.66, 495.32±13.96, and 339.92±11.22, respectively (F=1634.26, P<0.01). The one way analysis of variance showed significant differences in these parameters among groups. Conclusions: Corneal epithelial cells of rats activated by MNCs had higher CD80 expression, but the expression became decreased with micromolecular compound J2. The ultrastructure of the corneal epithelium became coarser after MNCs activation, and the number of protrusions increased significantly. The ultrastructural changes were alleviated by J2.(Chin J Ophthalmol, 2021, 57: 608-613).

DNA Methylation Differences in Peripheral Blood of Patients with Anaphylaxis.

ABSTRACT: Objective To study the DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and to provide a new research direction and basis for the forensic diagnosis of shock caused by drug hypersensitiveness. Methods Methylation microarray was used to detect DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and normal subjects. Sequencing data and chip data were analyzed for differences in DNA methylation using R language methylkit, ChAMP package. Random forest algorithm was used to evaluate the importance of the DNA methylation differential sites. Results Differential sites of DNA methylation highly associated with anaphylaxis caused by cephalosporin drugs were obtained at loci such as ETS1, PRR23B and GNAS. Conclusion Cephalosporin allergy is associated with DNA methylation, and DNA methylation may be a new strategy for forensic identification of anaphylactic shock and death.

Protein kinase Mζ in medial prefrontal cortex mediates depressive-like behavior and antidepressant response.

Neuronal atrophy and alterations of synaptic structure and function in the medial prefrontal cortex (mPFC) have been implicated in the pathogenesis of depression, but the underlying molecular mechanisms are largely unknown. The protein kinase Mζ (PKMζ), a brain-specific atypical protein kinase C isoform, is important for maintaining long-term potentiation and storing memory. In the present study, we explored the role of PKMζ in mPFC in two rat models of depression, chronic unpredictable stress (CUS) and learned helplessness. The involvement of PKMζ in the antidepressant effects of conventional antidepressants and ketamine were also investigated. We found that chronic stress decreased the expression of PKMζ in the mPFC and hippocampus but not in the orbitofrontal cortex. Overexpression of PKMζ in mPFC prevented the depressive-like and anxiety-like behaviors induced by CUS, and reversed helplessness behaviors. Inhibition of PKMζ in mPFC by expressing a PKMζ dominant-negative mutant induced depressive-like behaviors after subthreshold unpredictable stress and increased learned helplessness behavior. Furthermore, stress-induced deficits in synaptic proteins and decreases in dendritic density and the frequency of miniature excitatory postsynaptic currents in the mPFC were prevented by PKMζ overexpression and potentiated by PKMζ inhibition in subthreshold stress rats. The antidepressants fluoxetine, desipramine and ketamine increased PKMζ expression in mPFC and PKMζ mediated the antidepressant effects of ketamine. These findings identify PKMζ in mPFC as a critical mediator of depressive-like behavior and antidepressant response, providing a potential therapeutic target in developing novel antidepressants.

[Comparison of invasive and non-invasive blood pressure in critically ill patients].

Objective: To investigate the difference of invasive and noninvasive blood pressure in systolic, diastolic and mean blood pressure (SBP, DBP and MAP) levels in critically ill patients. Methods: A total of 197 critically ill patients were enrolled in the intensive care unit of Peking Union Medical College Hospital in this prospective observative study. There were 113 males and 84 females with a mean age of (59±17) years. With Pearson correlation and Bland-Altman analysis, the difference and agreement of invasive and noninvasive blood pressure in SBP, DBP and MAP levels were investigated. Results: Invasive blood pressure monitoring was significantly correlated with noninvasive blood pressure (the correlation coefficients of SBP, DBP and MAP was 0.908, 0.861 and 0.888, respectively, all P<0.001). The mean bias and 95% confidence interval (CI) in SBP(invasive)-SBP(noninvasive), DBP(invasive)-DBP(noninvasive), MAP(invasive)-MAP(noninvasive) respectively was as following: SBP (14.5±11.9) mmHg (-8.8-37.8 mmHg); DBP (1.0±7.2) mmHg(-13.2-15.2 mmHg); MAP (10.5±8.3) mmHg (-5.9-26.8 mmHg). Moreover, there was a significant positive correlation between the bias of SBP(invasive)-SBP(noninvasive) and the value of SBP; when the SBP(invasive)≥143 mmHg was used to predict a value of bias≥20 mmHg, the sensitivity was 72.4% and specificity was 81.3%, and the area under the receiver operating characteristic curve was 0.808 (95% CI: 0.745-0.870). Conclusions: There is a good agreement between invasive and noninvasive blood pressure in critically ill patients, the bias of invasive-noninvasive blood pressure measurement is positively correlated to the value of blood pressure.

[Effect of Δ40p53 isoform on enhancing the pro-apoptotic function of p53 in tumor cells].

Objective: To investigate the effect of Δ40p53, an alternative spliced isoform of p53 lacking the N-ter minus, on the pro-apoptotic function of p53. Methods: The wild-type p53 was ectopically expressed in HCT116-p53(-/-) (endogenous Δ40p53 expression), HCT116-p53(+ /+) (wild-type p53) and H1299 (p53-null) cells by adenoviral delivery, while Δ40p53 plasmid were transfected into these cells to overexpress Δ40p53. The levels of Δ40p53 and p53 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR. The expression of related proteins was deter mined by Western blotting. The interaction of p53 and Δ40p53 was observed by co-immunoprecipitation assay. Calcein-AM/propidium iodide (PI) staining and flow cytometry were used to detect the apoptotic rate of tested cells in each group. Results: HCT116-p53(-/-) cells expressed endogenous Δ40p53 isoform. Neither transcription nor protein expression of wild-type p53 was interfered by the increased expression of Δ40p53. Full length p53 and Δ40p53 could bind to each other. Calcein-AM/PI staining showed that the apoptotic rates of H1299-Control, HCT116-p53(-/-) -Control, H1299+ p53, HCT116-p53(-/-)+ p53, H1299+ oxaliplatin (Oxa), HCT116-p53(-/-)+ Oxa, H1299+ p53+ Oxa and HCT116-p53(-/-)+ p53+ Oxa groups were (2.50±0.47)%, (2.40±0.32)%, (5.20±0.58)%, (4.10±0.18)%, (22.40±1.73)%, (19.30±1.11)%, (29.90±1.15)% and (39.30±2.26)%, respectively. It was statistically significant between H1299+ p53+ Oxa and HCT116-p53(-/-)+ p53+ Oxa groups (t=3.721, P=0.0205). Moreover, the apoptotic rates of H1299-Control, H1299+ Δ40p53, H1299+ p53, H1299+ p53+ Δ40p53, H1299+ Oxa, H1299+ Δ40p53+ Oxa, H1299+ p53+ Oxa and H1299+ p53+ Δ40p53+ Oxa groups were (2.60±0.35)%, (2.20±0.17)%, (4.80±0.49)%, (4.90±1.10)%, (20.30±1.10)%, (19.60±1.45)%, (27.90±1.39)%, (35.20±1.43)%, respectively. Furthermore, flow cytometry assay showed that the apoptotic rates of above cells were (2.70±0.32)%, (2.20±0.24)%, (4.60±0.48)%, (3.90±0.67)%, (19.30±1.11)%, (17.70±0.66)%, (28.30±2.76)% and (37.50±1.51)%, respectively. H1299+ p53+ Δ40p53+ Oxa cells showed higher cell apoptosis than H1299+ p53+ Oxa cells (t=2.930, P=0.042). Conclusion: Δ40p53 isoform can bind to full-length p53, and enhance its pro-apoptotic function in tumor cells.

[Investigation of doctors' and nurses' perceptions and implementation of delirium management in intensive care unit].

Objective: To investigate doctors' and nurses' perceptions and implementation of delirium management in intensive care unit. Methods: A total of 197 doctors and nurses in 2 general ICUs and 3 special ICUs at Peking Union Medical College Hospital finished a self-designed questionnaire of delirium management. Results: There were 47 males and 150 females, 43 doctors and 154 nurses who participated in the survey.One hundred and twenty five participators were from general ICU and the others from special ICU. The ICU staff had a significant difference on the perceptions and implementation of delirium management(P< 0.001) including. Doctors and nurses scored lowest in "family engagement and empowerment" and "sleep management" with (4.620±2.393) and (5.430±2.153) respectively. There was a significant difference between nurses and doctors in the management of analgesia and sedation (P< 0.05).Doctors and nurses from different ICUs had significant discrepancy in the implementation of "delirium assessment" (P< 0.05). Conclusions: The ICU staff should improve the perceptions and the implementation of delirium management,especially in special ICUs. Delirium management should be included as a routine care in ICU to improve patients' outcome.

[Efficacy and safety of vertebrae pedicle subtraction osteotomy by posterior trans apical for correction of severe and rigid idiopathic scoIiosis].

Objective: To evaluate the efficacy and safety of pedicle subtraction osteotomy for correction of severe rigid idiopathic scoliosis. Methods: From January 2003 to December 2014, eighteen patients with severe rigid idiopathic scoliosis were treated by posterior wedge osteotomy in department of spinal surgery, the First Affiliated Hospital of Xinjiang Medical University. There were 5 males and 13 females, and the average age at was (16.9±9.1)years, and the average follow-up was 47.6 months. The coronal and sagittal Cobb's angles, sagittal trunk shifts, apical vertebral translation, height change and fusion segments were measured on standing AP and lateral radiographs before, after surgery and the final follow-up. Results: The Cobb's angle in the coronal plane was corrected from (94.3±13.3)°to (31.6±14.8)°, representing 64.8% correction rate. The Cobb's angle in the sagittal plane was corrected from (62.5±17.8)°to (27.1±11.7)°, representing 82.6% correction rate. Coronal trunk shift was obviously improved from -29.8 mm to -3.5 mm. the average fusion segments was 13.7±1.8 ranging from 11 to 17 segments. Perioperative complications consisted of pleura injury in 4 cases, cerebrospinal fluid leakage in 2 cases transient neurological damage in 2 cases and delayed infection in 1 case. Conclusion: Pedicle Subtraction Osteotomy correction by the single posterior approach is a reliable and safe surgical technique for correction of severe rigid idiopathic scoliosis.

Perceptions of public attitudes towards persons with mental illness in Beijing, China: results from a representative survey.

OBJECTIVES: Many studies have examined the general public's attitudes towards people with mental illness, but such studies are scarce in China. This study examined the perceptions of the Beijing population regarding their society's prevalent attitudes towards people with mental illness. METHODS: A total of 5000 individuals aged 18 or above living in Beijing were selected using a multistage, stratified, cluster and random sampling method. This was followed by a face-to-face interview which used a standardized questionnaire asking about societal attitudes towards individuals with mental illness. RESULTS: 4602 out of 5000 eligible individuals met the inclusion criteria and participated in the interview. 4596 questionnaires were deemed valid and included in the analyses. A large proportion of respondents believed that most individuals within their society held negative attitudes and had a strong desire to distance themselves from people with mental illness. Respondents aged 60 or older, who lived farther to downtown Beijing, or with higher education tended to believe that most individuals have relatively positive and tolerant attitudes towards people with mental illness. CONCLUSIONS: Many people in Beijing perceive that most members of their society have negative beliefs towards people with mental illness. Further efforts are needed to determine if these perceptions are accurate and to reduce the stigma that is reinforced by these perceptions.

[Diagnosis and treatment of Tuberculous primary psoas abscess].

Objective: To retrospective analysis the clinical characteristics of diagnosis and treatment the disease of tuberculous psoas abscess (TPA). Method: The medical records of 13 consecutive patients presenting to our hospital with TPA were reviewed from November 2002 to December 2015.Demographic data, presenting features, predisposing factors and the investigations performed were recorded and the outcomes were analysed. Results: The thirteen patients were included, 11 cases all had lower backache, 6 cases had Lower abdomen or groin area pain, 2 cases had fever, eleven cases were treated with Open drainage, one with Ultrasound guided drainage, and the last case used antibiotics alone, median hospital stay of 35 days, follow-up showed no recurrence or death in patients. Conclusion: The incidence of TPA is probably underreported.The vague presentation leads to delays in diagnosis and increases morbidity and a high index of suspicion is the key to early diagnosis.Open drainage has Lower recurrence and death rates, and used widely, is the most reliable method for the treatment of primary tuberculous psoas abscess.

Association analysis of yield and fiber quality traits in Gossypium barbadense with SSRs and SRAPs.

Cotton is an important cash crop. Mining for quantitative trait loci related to yield and fiber quality traits using association analysis has many advantages for cotton research. In this study, 170 simple sequence repeats (SSRs) and 258 sequence-related amplified polymorphisms (SRAPs) were used to analyze the association of 3 yield component traits and 5 fiber quality traits of 55 Gossypium barbadense accessions in 2009 and 2010. Principal component analysis of SSRs and SRAPs showed 3 and 2 subgroups, respectively. The boundaries between the SRAP groups were much more defined than those of the SSRs. A mixed linear model was used to analyze association of yield and fiber quality traits with SSRs and SRAPs. A total of 72 loci were detected, including 28 loci of SSRs and 44 loci of SRAPs; 26 of these loci were related to yield component traits, and 46 of these loci were related to fiber quality traits. The mean phenotypic variations explained in the SSR and SRAP analysis were 8.89 and 8.61%, respectively. The locus with the highest phenotypic variation explained was NAU1164 (23.33%), which was related to fiber uniformity. The comparison of association results between the two datasets showed that mining quantitative trait loci using association analysis was more efficient with SRAPs than with SSRs.

[Construction and pathological characterization of 3 animal models of temporomandibular joint degenerative joint disease in mice].

Objective: To explore the pathological characteristics of three mice models of temporomandibular joint degenerative joint disease (TMJDJD), including osteoarthritis and osteoarthrosis, and to provide references for animal experimental study regarding the pathological mechanism of osteoarthritis and osteoarthrosis. Methods: A total of 54 8-week-old male C57BL/6 mice were selected to construct three TMJDJD animal models, including bilateral temporomandibular joint (TMJ) Freund's complete adjuvant (FCA) injection model, bilateral TMJ monosodium iodoacetate (MIA) injection model, and right TMJ discectomy model. FCA injection model (15 mice) was divided into saline injection group, FCA injection group-1 week, FCA injection group-2 week, FCA injection group-4 week and FCA injection group-6 week, 3 mice were used at each time point, with a total of 6 TMJs on both sides. MIA injection model (15 mice) was separated into saline injection group, MIA injection group-1 week, MIA injection group-2 week, MIA injection group-4 week and MIA injection group-6 week, 3 mice were used at each time point, with a total of 6 TMJs on both sides. TMJ discectomy model (24 mice) was split into control group, discectomy group-2 week group, discectomy group-4 week and discectomy group-6 week, six mice were used at each time point, with a total of six right TMJs. General pictures of the bilateral joints area of mice were collected 1 day after drug injection, and stereoscopic images of condylar tissues were collected 4 weeks after microsurgery for discectomy. Mouse TMJ tissue sections from each time point were stained with HE and toluidine blue, respectively, synovial tissues were scored for synovial inflammation, and the percentage of proteoglycan in condylar cartilage was quantitatively analyzed. Results: One day after intra-articular FCA or MIA injection, the width of bilateral TMJ were significantly increased in FCA injection groups [(24.60±0.46) mm] compared with the saline injection group [(21.63±0.52) mm] (t=4.25, P<0.013), the width of bilateral TMJ in MIA injection groups [(24.50±0.62) mm] were also significantly higher than that in saline injection group [(21.40±0.52) mm] (t=3.82, P=0.019). The synovitis scores in FCA injection groups 1, 2, 4, 6 weeks after FCA injection were significantly higher than that of the saline injection group (F=18.09, P<0.001), with the proteoglycan of condylar cartilage increased firstly and then decreased compared with the saline injection group (F=21.59, P<0.001). Condylar cartilage proteoglycan loss in different degrees were observed 1, 2, 4 and 6 weeks after MIA injection (F=13.59, P<0.001), and synovitis scores were increased at different degrees compared with saline injection group (F=14.79, P<0.001). The morphology of condylar cartilage in discectomy groups mice were severely damaged, synovial tissues showed dense connective tissue lesions at 2, 4 and 6 weeks postoperatively, condylar cartilage tissues showed a time-dependent loss of proteoglycan compared with the control group (F=40.62, P<0.001). Conclusions: Intra-articular FCA injection establishes a mouse model of TMJ osteoarthritis with severe synovial inflammation. Intra-articular MIA injection constructs a mouse model of typical TMJ osteoarthritis. Discectomy establishes a mouse TMJ osteoarthrosis model with severe condylar cartilage destruction.

Expression of miR-182 in patients with fracture of tibial plateau and its regulative effects on the fracture healing.

OBJECTIVE: To investigate the expression of miR-182 in patients with fracture of tibial plateau (FTP) and its effects on osteoblasts and fracture healing. PATIENTS AND METHODS: The patients with fracture of tibial plateau treated in our hospital and healthy subjects who received physical examination from January 2017 to January 2018 were collected. The expression of miR-182 in the serum was detected. The osteoblasts from SD rats were cultured and transfected with miR-182, anti-miR-182, miR-NC or anti-miR-NC using transfection reagent LipofectamineTM 2000. The proliferation, alkaline phosphatase (ALP) activity, calcification and osteogenic gene expression of osteoblasts were detected. The rat models with fracture of tibial plateau were divided into control group, fracture group, fracture+miR-182 group, and fracture+anti-miR-182 group. The levels of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and transforming growth factor ß (TGFß) in serum were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with the controls, the expression of miR-182 in serum was significantly elevated in patients with fracture of tibial plateau. Overexpression of miR-182 inhibited the proliferation of osteoblasts, while the knockdown of miR-182 increased the proliferation. MiR-182 could decrease the ALP activity of osteoblasts, while anti-miR-182 increased the ALP activity. Osteoblast calcification ability was significantly decreased by overexpression of miR-182. Knockdown of miR-182 increased the calcification ability of osteoblasts and the expression of osteogenic genes. MiR-182 could inhibit the expression of osteogenic genes. The levels of VEGF, EGF and TGFß in the fracture group were higher than those in the control group, while the levels in the fracture+miR-182 group were higher than those in the fracture group. The levels of VEGF, EGF and TGFß in the anti-miR-182 group were lower than those in the fracture group. CONCLUSIONS: MiR-182 is elevated in patients with fracture of tibial plateau, which can inhibit the proliferation and differentiation of osteoblasts and affect the fracture healing. The knockdown of miR-182 might be a new method for treating fracture healing.

Long non-coding RNA OIP5-AS1 plays an oncogenic role in ovarian cancer through targeting miR-324-3p/NFIB axis.

OBJECTIVE: Long non-coding RNAs (lncRNAs) have been found to exert specific functions in the progression of ovarian cancer (OC), except for lncRNA-OIP5-AS1. In this study, we aim at exploring the molecular mechanisms of OIP5-AS1 in OC. PATIENTS AND METHODS: The expression levels of OIP5-AS1, miR-324-3p, and NFIB in OC tissues and OC cell lines were explored by qRT-PCR assay. The OC cell vitality was examined by CCK-8 and transwell assay. The protein expression level of NFIB was measured by Western blot analysis. The correlation between OIP5-AS1, miR-324-3p, and NFIB was appraised by Dual-Luciferase reporter assay. RESULTS: OIP5-AS1 and NFIB were validated to be upregulated in both OC tissues and OC cell lines. Inversely, miR-324-3p downregulation was found in OC tissues and OC cell lines. Functionally, OIP5-AS1 knockdown and miR-324-3p overexpression restrained SKOV3 cell viability, invasion, and migration. Our results verified that OIP5-AS1 inhibited the expression of miR-324-3p in OC. Moreover, miR-324-3p directly targets NFIB. Besides that, NFIB silencing restrained the progression of SKOV3 cells. CONCLUSIONS: The present study clarified that OIP5-AS1 accelerated OC progression by sponging miR-324-3p and upregulating NFIB. OIP5-AS1 can be a possible therapeutic target for the treatment of OC.

The medial compartment and patellofemoral joint degenerate more severely in early stage knee osteoarthritis: a cross-sectional study.

OBJECTIVE: The purpose of this study was to compare the differences of degenerative characteristics of medial, lateral regions, femoral, patellar, and tibia parts of the knee joint in the early stage of knee osteoarthritis (OA). PATIENTS AND METHODS: A total of 66 patients with early-stage knee OA and 22 healthy volunteers who have no knee-related clinical symptoms were enrolled in this cross-sectional study. T2 mapping and 3D dual-echo images were acquired with a 3.0T MR scanner. The degenerative changes of the articular cartilage were quantified by a T2 mapping and cartilage thicknesses analysis. Any structural changes were conducted using the Whole Organ Magnetic Resonance Imaging Score (WORMS). RESULTS: In patients with knee OA, the thicknesses of medial cartilages were significantly thinner than lateral ones (2.13 mm vs. 2.34 mm, p<0.0001), but with higher T2 values (40.38 ms vs. 38.4 ms, p<0.0001) and WORMS scores (12.12 vs. 0.47, p=0.004). No significant differences were observed between medial and lateral cartilage in the healthy volunteers. The T2 values of the femoral (p<0.001) and patellar (p=0.012) cartilages of OA patients were higher than that of the healthy controls. Within OA group, the T2 values of femoral (p<0.0001) and patellar (p<0.0001) cartilages were higher than tibial ones. Moreover, the WORMS scores of femoral (p=0.001) and patellar (p<0.0001) cartilages were higher than that of the tibial ones. CONCLUSIONS: This study demonstrates that the medial compartment and patellofemoral knee joint degenerate more severely in patients with early-stage knee OA.

LINC00472 promotes osteogenic differentiation and alleviates osteoporosis by sponging miR-300 to upregulate the expression of FGFR2.

OBJECTIVE: To investigate the expression of LINC00472 in osteoporotic issues of patients, ovariectomized (OVX) mice and mice bone marrow mesenchymal stem cells (BMSCs), its effect on osteogenic differentiation of BMSCs and its mechanism. PATIENTS AND METHODS: The expression of LINC00472 and miR-300 in osteoporosis patients (n=55), ovariectomized (OVX) mice (n=10) and mice BMSCs (n=3) was detected by RT-qPCR and the correlation between the expression of miR-300 and LINC00472 was analyzed. After transferring sh-LINC00472 and overexpression LINC00472 plasmids into mice BMSCs, the expression of ALP, Bglap, OPN, Runx2 was detected by RT-qPCR and Western blot, which were related with osteogenic differentiation. In addition, Luciferase activity was used to detect whether miR-300 combined with LINC00472 and FGFR2 in mice BMSCs directly. Finally, Western blot (WB) was used to detect the change of FGFR2 protein expression by miR-300 inhibitor and sh-LINC00472. RESULTS: We found there was a negative correlation between the expression of miR-300 and LINC00472 in osteoporosis patients, bone tissues of OVX mice and mice BMSCs. The expression of LINC00472 in mice BMSCs was gradually increased with osteogenic differentiation. Transferring overexpression plasmid of LINC00472 into BMSCs, the expression of ALP, Bglap, OPN, Runx2 was increased both in mRNA and protein levels. Transferring sh-LINC00472 to BMSCs, the results were the opposite. Luciferase results showed that miR-300 could directly bind to LINC00472 and FGFR2 in mice BMSCs. What's more, RT-qPCR and WB results showed that transferring sh-LINC00472 could decrease the expression of FGFR2 mRNA and protein, while miR-300 inhibitor could recover this tendency. CONCLUSIONS: According to these results, this study revealed the previously neglected LINC00472/miR-300/FGFR2 regulatory axis for the regulation of osteogenic differentiation in osteoporosis, which may be a potential target for the treatment of osteoporosis.

The phosphorylation of hCDC14A modulated by ZIPK regulates autophagy of murine pancreatic islet ß-TC3 cells upon glucose stimulation.

OBJECTIVE: Depletion of islet ß cells plays a crucial role in the onset of diabetes mellitus. Cell autophagy, as a self-healing process, contributes to maintaining metabolic homeostasis and can protect islet ß cells from apoptosis upon starvation or high glucose stress. However, the underlying regulatory network of the autophagic process in islet ß cells has not been fully explored. MATERIALS AND METHODS: Murine ß-TC3 cells treated with different concentrations of glucose, and wild-type or the Ser484 mutant human cell division cycle gene 14A (hCDC14A) was transfected. Cell viability, proliferation and autophagy as well as islet secretion were studied. The mTOR and AMPK signaling pathways were investigated by western blots. Zipper-interacting protein kinase was studied using mass spectrometry and immunoprecipitation. RESULTS: Overexpression of wild-type hCDC14A, but not the Ser484 mutant hCDC14A, promoted cell viability, proliferation and autophagy accompanied by enhanced islet secretion and reduced cell apoptosis via mTOR pathway inhibition as well AMPK pathway activation in ß-TC3 cells and vice versa. Furthermore, Zipper-interacting protein kinase (ZIPK), also known as DAPK3, was found to interact with hCDC14A primarily for Ser484 phosphorylation, and ZIPK knockdown could affect the phosphorylation of hCDC14A and weaken cell death or cell cycle modulation. CONCLUSIONS: Taken together, our results may provide new insight into the role of hCDC14A in the autophagy of islet ß cells and suggest the potential therapeutic value of hCDC14A phosphorylation in the prevention and treatment of diabetes.

[Split-thickness scalp and allogenic acellular dermal matrix in repairing deep wounds of hands in patients with extremely extensive burns].

Objective: To explore the clinical effects of split-thickness scalp and allogenic acellular dermal matrix (ADM) in repairing deep wounds of hands in patients with extremely extensive burns. Methods: Six patients with extremely extensive burns complicated by deep burn in hands were admitted to our department from December 2014 to December 2017, including 4 males and 2 females, aged 21 to 58 years. Their total burn areas were from 85% to 95% total body surface area (TBSA), and the sum of deep-partial thickness and full-thickness wounds was larger than 50% TBSA. Under general anesthesia, deep wounds of 10 dorsal hands were repaired by combined transplantation of split-thickness scalp and allogenic ADM in 4 to 6 weeks after injury. The skin grafting range was beyond the metacarpophalangeal joint, including partial dorsal digital deep wounds. Survival of skin grafts on dorsal hands, follow-up of hand shape and functional recovery were observed and recorded. Results: Six patients were successfully treated. There was no infection on the dorsal deep wounds of 10 hands after surgery, and the grafts survived well. During follow-up of 1 to 2 years after operation, there were no recurrent tension blisters or dorsal extension deformity of the metacarpophalangeal joints in 10 hands, the shape and function of hands recovered well, and hand Carroll scores were from 90 to 99 points, with functional levels of Ⅴ and Ⅵ. Conclusions: The combined transplantation of split-thickness scalp and allogenic ADM is a good method for repairing deep hand wounds in patients with extremely extensive burns, which can alleviate the cicatrix hyperplasia and contracture of healed hand wounds, and improve the shape and function of hands.

Radioprotective gene therapy.

Control of cancer by irradiation therapy alone or in conjunction with combination chemotherapy is often limited by organ specific toxicity. Ionizing irradiation toxicity is initiated by damage to normal tissue near the tumor target and within the transit volume of radiotherapy beams. Irradiation-induced cellular, tissue, and organ damage is mediated by acute effects, which can be dose limiting. A latent period follows recovery from the acute reaction, then chronic irradiation fibrosis (late effects) pose a second cause of organ failure. We have developed the technology for radioprotective gene therapy using the transgene for the antioxidant manganese superoxide dismutase, delivered to specific target organs (lung, esophagus, oral cavity, oropharynx, and bladder) using gene transfer vectors including plasmid/liposomes (PL) and adenovirus. Irradiation protection by MnSOD transgene overexpression at the cellular level has been demonstrated to be localized to the mitochondrial membrane. Using MnSOD transgene constructs lacking the mitochondrial localization leader sequence, and in other experiments attaching this localization signal to otherwise non-radioprotective cytoplasmic Cu/ZnSOD, mitochondrial localization has been demonstrated to be critical to protection. Organ specific injection of MnSOD-PL prior to irradiation demonstrates transgene expression for 48-72 hours, and an associated decrease in ionizing irradiation-induced expression of inflammatory cytokine mRNA and protein. Significant reduction of organ specific tissue injury has been demonstrated in several organ systems in rodent models. Application of MnSOD-PL gene therapy in the setting of fractionated chemo-radiotherapy is being tested in clinical trials for prevention of esophagitis during treatment of non-small cell carcinoma of the lung, and in prevention of mucositis during combination therapy of carcinomas of the head and neck. Encouraging results in pre-clinical models suggest that radioprotective gene therapy may facilitate dose escalation protocols to allow increases in the therapeutic ratio of cancer radiotherapy.

Lipogenesis stimulatory and inhibitory activities of the insulin mediators.

Incubating plasma membranes prepared from pig liver with varying concentrations of insulin (50-1000 microU/ml) resulted in the release of at least two insulin chemical mediators. One of them was fraction 1 of insulin mediator (M. W. 3700-4000 daltons) which had a significant lipogenesis-stimulating activity. The other was fraction 2 of insulin mediator (M. W. about 1000 daltons) which exhibited a lipogenesis-inhibitory activity. The ratio of yield between the two mediators produced from the membranes was not only dependent on the concentration but also on the potency of insulin and its analogs added. The result showed that there was more production of fraction 2 than fraction 1 with the inducer at low concentration (100 microU/ml), while the production of fraction 1 from the plasma membranes incubated with high concentration of insulin (300 microU/ml) was higher than fraction 2. On the other hand, insulin and its analogs which have different biological activities and receptor binding activities have been used to induce the insulin mediators. The results obtained were similar to those mentioned above. This suggested that the generation of the mediators was dependent on the biological potences but not the binding activities.