High dose insulin promotes the proliferation of vascular smooth muscle cells via AP-1/SM-α pathway.

Proliferation of vascular smooth muscle cells (VSMCs) participates in multiple cardiovascular disorders, while the mechanism remains unclear. This study aims to investigate the effects of insulin on VSMC. Insulin was used to stimulate rat VSMCs, and the effects on cell cycle and proliferation were subsequently analyzed using flow cytometry. Furthermore, AP-1 and SM-α overexpression vectors were constructed and transfected into VSMCs. AP-1 and SM-α were inhibited by SR11302 and SM-α siRNA, respectively. The mRNA and protein expression levels were subsequently detected using the reversetranscription quantitative polymerase chain reaction and western blotting, respectively. AP-1 and SM-α gene promoter binding sites were determined using luciferase and chromatin immunoprecipitation assays. As a result, we found that high dose of insulin promoted proliferation of VSMCs and increased the percentage of cells in the S phase by downregulating AP-1. AP-1 was identified to bind to the SM-α gene promoter at locus 2-177 to upregulate SM-α gene expression. Inhibition of AP-1 led to the decrease of SM-α expression. Overexpression of SM-α directly suppressed proliferation of VSMCs, while knocking it down promoted the process. Therefore, this study revealed that insulin downregulated the expression of the SM-α gene by inhibiting AP-1, which in turn facilitated proliferation of VSMCs.

[Impact of health literacy on patient experience of outpatients in China and its mechanism].

OBJECTIVE: To understand the relationship between health literacy and patient experience of outpatients in China, and to explore its mechanism. METHODS: The conceptual framework was developed based on Andersen's behavioral model of health services use and health literacy skills framework. An online cross-sectional survey was conducted with snowball sampling method, while the health literacy was measured by self-designed patient health literacy scale, and the patient experience was measured by the Chinese patient experience questionnaire for ambulatory care developed by Peking Union Medical College. And a structural equation model was built to explore the relationship between them and test the mechanism of health literacy influencing patient experience. RESULTS: A total of 2 773 subjects were investigated. The average score of health literacy was (90.72±12.90) points, accounting for 78.89% of the full score, and the dimension of seeking social support had the lowest score. The average score of overall rating of patient experience was (3.71±0.74) points, and the scores of each dimension of patient experience were between 3.56 and 3.80. The model fit indices of structural equation model for overall rating of patient experience among the outpatients were χ2/df=9.29 (χ2=4 107.27, df=442), root mean square error of approximation (RMSEA)=0.055 (< 0.06), comparative fit index (CFI)=0.926 (>0.90), Tucker-Lewis index (TLI)=0.918 (>0.90), standardized root mean square residual (SRMR)=0.061 (< 0.08), the model was acceptable. The variance in patient experience explained by the model was 0.108. The structural equation model analysis results showed that the overall rating of outpatient experience was directly affected by health literacy (ß=0.263, P < 0.001), also indirectly affected by health literacy (ß=0.012, P < 0.001). In other words, the overall rating increased by 0.275 units for each standard deviation increase of health literacy. Self-evaluated health status mediated the relationship between health literacy and the overall rating of outpatient experience. In terms of diffe-rent dimensions of patient experience, the standardized path coefficient of the total effect of health literacy on patient experience was as follows: Information guidance 0.337, humanistic care 0.319, communication with doctors 0.294, service efficiency 0.240, and hospital environment 0.173. CONCLUSION: The patients with higher level of health literacy were more likely to have a better outpatient experience in China, and the information guidance experience and humanistic care experience were most affected by health literacy. And the communication and information utilization ability had the greatest influence on patient experience.

[Effect of continuous blood purification for acute renal injury after acute Stanford type A aortic dissection].

Objective: To investigate the effect of continuous renal replacement therapy (CRRT) on acute kidney injury (AKI) after acute Stanford type A aortic dissection (ATTAD). Methods: In this study, 120 patients with AKI after ATTAD surgery treat in Gansu Provincial People's Hospital were selected as research objects. Among them, there were 86 males (71.7%) and 34 females (28.3%) with a mean age of (55±5) years. These patients were randomly divided into experimental group (n=60) and control group (n=60) with stratified random sampling. CRRT and intermittent hemodialysis (IHD) were performed in the experimental group and the control group respectively. The therapeutic effect of CRRT on ATTAD patients with AKI was evaluated by blood purification index, renal function index, Sequential Organ Failure Assessment (SOFA) score, inflammatory level, hemodynamic index and fluid infusion volume. Results: The two treatment schemes both had considerable therapeutic effects on the condition of patients, but the therapeutic effect of CRRT was more superior. In the patients treated with CRRT, the levels of serium creatinine (SCr), blood urea nitrogen (BUN) and blood lactic acid (Lac) were all lower than those in the control group (all P<0.05). The time of staying in intensive care units (ICU), the period of oliguria, the times of renal replacement therapy, the time from the first dialysis to the last dialysis and the total hospital stay in the experimental group were all shorter than those in the control group (all P<0.05). The volume of fluid infusion was less and the hemodynamic index was better than that in the control group, but there was no significant difference in hospital mortality between the two groups (P>0.05). The levels of interleukin (IL)-6, IL-8 and C-reactive protein (CRP) in the experimental group were (21.9±1.8) ng/L, (18.6±1.4) ng/L and (22.7±2.2) mg/L, respectively, which were all significantly lower than those in control group ((27.9±3.2) ng/L, (28.3±1.4) ng/L, (60.1±2.5)mg/L, respectively; t=14.527, 13.255, 11.247, all P<0.05). The scores of SOFA at all time points in the experimental group were all lower than those in the control group (all P<0.05). Conclusion: Compared with IHD, CRRT brings no significant reduction in hospital mortality in patients with AKI after ATTAD, but shows better prognosis.

[Role of growth arrest-specific protein 6 in migration and osteogenic differentiation of human periodontal ligament cells].

OBJECTIVE: To investigate the role of growth arrest-specific protein 6 (Gas6) in the process of the migration and osteogenic differentiation of human periodontal ligament cells (hPDLCs). METHODS: After different concentrations of recombinant human Gas6 (rhGas6) were added to hPDLCs, cell prolife-ration experiment (CCK-8) was taken to observe the effect of rhGas6 on hPDLCs cell proliferation. Scratch test and cell migration test (Transwell) were taken to analyze the migratory ability of hPDLCs in different concentrations of rhGas6 groups. After osteogenic induction, real-time quantitative polymerase chain reaction (real-time PCR) was taken to detect the expression of the Runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP). ALP staining was used to detect the amount of mineralized nodules. RESULTS: After adding different concentrations of rhGas6, there were no statistically significant differences in hPDLCs cell proliferation among the experimental groups and the control group at 24, 48 and 72 hours (P>0.05). After 24 h of scratch, the healing area in the 800 µg/L of the rhGas6 group was greater than that in the control group, but without statistically significant difference (31.06%±13.70% vs. 21.79%±9.51%, P>0.05). In the migration test, after 24 h, the number of hPDLCs cells which penetrated through the membrane in the 800 µg/L rhGas6 group was significantly higher than that in the control group (P < 0.01). After rhGas6 was added and osteogenic induction, Runx2 and ALP gene expressions of hPDLCs in the 800 µg/L group were significantly higher than those in the control group (1.60±0.30 vs. 0.91±0.10, 2.81±0.61 vs. 0.86±0.12, P < 0.01). After Gas6 was knocked down, the ALP expression of hPDLCs was significantly lower than that of the control group (0.39±0.07 vs. 0.92±0.14, P < 0.01). There was no significant change in Runx2 expression (P>0.05). After 7 days of osteogenic induction, the mineralized nodules formed in the Gas6 knockdown group were significantly less than those in control group (0.25±0.04 vs. 1.00±0.11, P < 0.001). After 14 days of induction, the staining degree of the Gas6 knockdown group was lower than that of the control group, but there was no significant difference (0.86±0.04 vs. 1.00±0.16, P>0.05). CONCLUSION: After downregulation of Gas6 gene, mineralized nodule formation was reduced and ALP gene expressions were decreased in the early stage of osteogenic induction (7 days). After addition of rhGas6, Runx2 and ALP gene expressions were increased and the number of cell migration was increased, suggesting that Gas6 might play a promoting role in the migration and osteogenic differentiation of human periodontal ligament cells.

[Determination of N-[4- (4-aminobenzyl) -phenyl] acetamide hemoglobin adduct in blood by ultra performance liquid chromatography-tandem mass spectrometry].

Objective: To establish the method for the determination of N-Acetyl-4, 4'-diaminodiphenylmethane (AcMDA) adduct in the hemoglobin by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) . Methods: The 20 mg hemoglobin sample was weighed into a 15 ml centrifuge tube, adding 20 µg/L internal standard solution AcMDA-D8 10 µl, then hydrolyzed with 1 ml 0.1 mol/L sodium hydroxide solution at 37°C for 0.5 hours, extracted with dichloromethane and concentrated by vacuum concentrator. The residue was dissolved in acetonitrile and detected by UPLC-MS/MS, then quantitative by internal standard method. Results: The linearity of the method was good at the range of 0.05-25.00 µg/L with a correlation coefficient of 0.9996, the detection limit (LOD) and limit of quantitation (LOQ) were 0.07 and 0.2 ng/g Hb, respectively. The recovery rate was ranged from 91.0%-95.4%; the relative standard deviation (RSD) of intra-and inter-batch precision were 4.5%-6.3% and 3.7%-4.4%, respectively. Conclusion: The determination method meet the requirement of GBZ/T 210.5-2008.

[Determination of tantalum and tantalum pentoxide in the air of workplace by inductively coupled plasma optical emission spectrometry].

Objective: To establish the method for determination of tantalum and tantalum pentoxide in the air of workplace by inductively coupled plasma optical emission spectrometry (ICP-OES). Methods: The tantalum and tantalum pentoxide in the workplace air were collected by filter membrane and then digested by microwave digestion apparatus in the mixed solvents (HNO(3)∶HF=3∶1) ,diluted to 50 ml and detected by ICP- OES. Results: The linearity of tantalum was good at the range of 0.0~100.0 µg/ml with a correlation coefficient of 0.9997, the LOD and LOQ were 0.04 µg/ml and 0.14 µg/ml, respectively. The recovery was ranged from 83.1%-87.8%; the RSD of intra-and inter-batch precision were 0.2%-1.0% and 1.1%-2.4%, respectively. The recovery was ranged from 87.4%-93.3%; the RSD of intra-and inter-batch precision were 0.5%-1.5% and 1.3%- 1.8%, respectively. Conclusion: The determination method meet the requirement of analysis and apply to the determination of tantalum and tantalum pentoxide in the air of workplace.

Simplified LC-MS/MS method for quantification of IG-105, a novel tubulin ligand, and its application to the pharmacokinetic study in rats at the anticancer effective dose.

IG-105, N-(2, 6-dimethoxypyridine-3-yl)-9-methylcarbazole-3-sulfonamide, a novel carbazole sulfonamide, shows a potent anticancer activity in a variety of human tumor cells in vitro and in vivo. In the present study, a rapid and convenient liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and applied to the pharmacokinetic study of IG-105 in rats. Chromatographic separation was accomplished on a C18 column using an isocratic mobile phase of acetonitrile-water-acetic acid (56:44:0.2, v/v/v). The ion transitions of IG-105 and combretastatin A4 (internal standard) in selected reaction monitoring mode were m/z 398→154 and m/z 317→286, respectively. The assay exhibited good linearity over the range of 2-512 ng/mL. Intra- and inter-day precisions were within 8.2 %, and the accuracies ranged from -6.0 to 3.7 %. The extraction recoveries were higher than 90 %, and the matrix effects were negligible. All quality control samples were stable at different storage conditions. The validated LC-MS/MS method was successfully applied to a preclinical pharmacokinetic study of IG-105 in rats after a single oral dose of 100, 250, or 1000 mg/kg which showed tumor growth inhibition activity. The absorption of IG-105 was proved to be rapid but saturated to a certain extent into the blood circulation, from where it was distributed and eliminated gradually.

[Selective tarsometatarsal arthrodesis for old Lisfranc injury].

Objective: To discuss the methods and clinical outcomes of selective tarsometatarsal (TMT) arthrodesis for old Lisfranc injury. Methods: The clinical data of 36 cases with old Lisfranc injury treated by selective arthrodesis from January 2010 to October 2016 in the Department of Orthopedics in Shanghai Sixth People's Hospital were analyzed retrospectively. There were 16 males and 20 females in this group with a mean age of (40±6) years. The information of pre-operative and post-operative X-ray, American Orthopaedics Foot and Ankle Society (AOFAS) midfoot score and pain Visual Analogue Scale (VAS) score was collected. The complications were also recorded. The pre- and post-operative data were compared with t test. Results: The 36 patients got a follow-up for at-least 2 years (averaged (4.3±1.6) years, ranged from 2 to 8 years). The post-operative AOFAS midfoot score was improved from (44±7)(28-60) to (83±7)(76-97)(t=-37.1, P<0.05), and the VAS score decreased from (6.3±2.5)(5-9) to (1.6±1.3)(0-3)(t=23.7, P<0.05). Implant breakage occurred in two patients and the symptom was relieved after the removal of implants. Conclusion: The selective TMT arthrodesis for old Lisfranc injury may relieve the symptoms, improve the function and life quality of patients by restoring the medial arch and midfoot and forefoot alignment.

[Determination of tungsten and insoluble compounds in the air of workplace by ICP-OES].

Objective: To establish the method for determination of tungsten and insoluble compounds in the air of workplace by inductively coupled plasma optical emission spectrometry (ICP-OES) . Methods: The tungsten and insoluble compounds were digested by microwave digestion apparatus using nitric acid and hydrofluoric acid, detected by ICP-OES. Results: The linearity of tungsten and insoluble compounds (as tungsten) were good at the range of 0.16-100.0 µg/ml, the minimum quantitation concentration was 0.11 mg/m(3), the recovery was ranged from 81.9%-97.1%, the RSD of intra-and inter-batch precision were 0.5%-2.5% and 1.2%-4.4%, respectively. Conclusion: The determination method meet the requirement of analysis and apply to the determination of tungsten and insoluble compounds in the air of workplace.

[Preliminary clinical evaluation of the esthetic effect of deep discolored anterior teeth restored with zirconia veneers].

OBJECTIVE: To observe the esthetic effect of deep discolored anterior teeth restored by zirconia veneers. METHODS: Small defected deep discolored anterior teeth with complete root canal therapy were restored by zirconia veneers (n=15). The same name teeth on the other side of the same dental arch were chosen as control teeth. The color difference values ΔE of the neck 1/3, the middle 1/3 and the incisor 1/3 between the deep discolored tooth and the normal control tooth before and after therapy were measured to evaluate the esthetic effect of zirconia veneer restoration. At the same time, the marginal fit of zirconia veneers was checked by the standard of United States Public Health Service (USPHS). The integrity of the veneers was also examined. RESULTS: On the labial side, fibers color difference values ΔE of the neck 1/3, the middle 1/3 and the incisor 1/3 between deep discolored teeth and normal control teeth were measured by the electronic colorimeter, which were 24.92±3.00,26.64±4.00 and 21.94±3.31 respectively. All the values were above 4.0, which were considered unacceptable in clinic. After restoration by zirconia veneers, the color difference values ΔE of the middle 1/3 and the incisor 1/3 between the restored and control teeth were 1.82±0.17 and 1.84±0.21. Both values were less than 2.0, which indicated both good color matching. The color difference value ΔE of the neck 1/3 was 3.92±0.48, which was less than 4.0 and could be accepted in clinic. The statistical analysis of the colors of the teeth before and after restoration compared with the control teeth was done by Paired t test. The t values in the neck 1/3, the middle 1/3 and the incisor 1/3 were 30.37, 21.56, 23.37 respectively. In the three group, all the P<0.001. There were obvious statistical differences. According to the standard of USPHS, the marginal fit of all the restored teeth was perfect (grade A). No zirconia veneers were broken or detached in the period of observation. CONCLUSION: Zirconia veneers can be a good method to restore deep discolored anterior teeth. However, it should be used cautiously when the patient's esthetic expectation was too high.

S-phase kinase-associated protein 2 expression interference inhibits breast cancer cell proliferation.

We investigated the expression of S-phase kinase-associated protein 2 (SKP2) in breast cancer tissues, and the effects of SKP2-specific small interfering RNA (siRNA) interference on breast cancer cell proliferation. Thirty subjects provided breast cancer tissue samples and 18 subjects provided normal breast specimens for this study. The expression of SKP2 in breast cancer patient tissues and normal breast tissues was detected by western blotting analysis and reverse transcription-polymerase chain reaction. SKP2-specific siRNA was used to decrease SKP2 expression in breast cancer cell line MDA-MB-231. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell proliferation. SKP2 expression in breast cancer tissues was significantly higher than in normal breast tissues (P < 0.05). Two pairs of siRNA specific to SKP2 were required to downregulate SKP2 expression in the breast cancer cell line MDA-MB-231. The MTT assay showed that MDA-MB-231 growth significantly slowed after SKP2 interference. Patients with breast cancer have an increased SKP2 level. Interference in SKP2 gene expression can inhibit breast cancer cell growth, suggesting that SKP2 is potentially a new target for breast cancer therapy.

Mitigated Tregs and augmented Th17 cells and cytokines are associated with severity of experimental autoimmune neuritis.

Experimental autoimmune neuritis (EAN), an animal model of human Guillain-Barré syndrome, has long been considered as a T helper (Th) 1 cell-mediated autoimmune disorder. However, deficiency of IFN-γ, a signature Th1 cytokine, aggravated EAN, with features of elevated production of IL-17A, despite an alleviated systemic Th1 immune response. We hypothesized that Th17 cells and their cytokines might play a pathogenic role in EAN. To further clarify the roles of these Th and regulatory T cell (Treg) cytokines in the pathogenesis of EAN and their interrelationship, we investigated the expression of Th1/Th2/Th17/Treg cytokines in EAN in this study. We found that the levels of Th17 cells and IL-17A in cauda equina (CE)-infiltrating cells and splenic mononuclear cells (MNCs) as well as in serum paralleled the disease evolution, which increased progressively during the initiation stage and reached higher value at the peak of EAN. The same pattern was also noticed for the expression of IL-22. The diverse expression profiles of FoxP3, IL-17 receptors A and C were seen in CE-infiltrating cells and splenic MNCs in EAN. These findings indicate a major pro-inflammatory role of Th17 cells and IL-17A in the pathogenesis of EAN. Therapeutic interventions may be focused upon inhibiting Th17 cells and their cytokines in the early phase of EAN, so as to delay and suppress clinical signs of the disease, which has relevance for future studies on pathogenesis and treatment of GBS in humans.

[Association between congenital hypothyroidism and in-hospital adverse outcomes in very low birth weight infants].

Objective: To investigate the association between congenital hypothyroidism (CH) and the adverse outcomes during hospitalization in very low birth weight infants (VLBWI). Methods: This prospective, multicenter observational cohort study was conducted based on the data from the Sino-northern Neonatal Network (SNN). Data of 5 818 VLBWI with birth weight <1 500 g and gestational age between 24-<37 weeks that were admitted to the 37 neonatal intensive care units from January 1st, 2019 to December 31st, 2022 were collected and analyzed. Thyroid function was first screened at 7 to 10 days after birth, followed by weekly tests within the first 4 weeks, and retested at 36 weeks of corrected gestational age or before discharge. The VLBWI were assigned to the CH group or non-CH group. Chi-square test, Fisher exact probability method, Wilcoxon rank sum test, univariate and multivariate Logistic regression were used to analyze the relationship between CH and poor prognosis during hospitalization in VLBWI. Results: A total of 5 818 eligible VLBWI were enrolled, with 2 982 (51.3%) males and the gestational age of 30 (29, 31) weeks. The incidence of CH was 5.5% (319 VLBWI). Among the CH group, only 121 VLBWI (37.9%) were diagnosed at the first screening. Univariate Logistic regression analysis showed that CH was associated with increased incidence of extrauterine growth retardation (EUGR) (OR=1.31(1.04-1.64), P<0.05) and retinopathy of prematurity (ROP) of stage Ⅲ and above (OR=1.74(1.11-2.75), P<0.05). However, multivariate Logistic regression analysis showed no significant correlation between CH and EUGR, moderate to severe bronchopulmonary dysplasia, grade Ⅲ to Ⅳ intraventricular hemorrhage, neonatal necrotizing enterocolitis in stage Ⅱ or above, and ROP in stage Ⅲ or above (OR=1.04 (0.81-1.33), 0.79 (0.54-1.15), 1.15 (0.58-2.26), 1.43 (0.81-2.53), 1.12 (0.70-1.80), all P>0.05). Conclusion: There is no significant correlation between CH and in-hospital adverse outcomes, possibly due to timely diagnosis and active replacement therapy.

Signal-processing electronics for stable and sensitive weak-field atomic vector magnetometers.

We present the electronics developed for a sensitive and stable atomic vector magnetometer used in low-field detections. These electronics are required to be not only highly reliable and sophisticated for signal processing but also compact in size and low cost in resource consumption for the purpose of miniaturization. In addition, this magnetometer works with multiple modulations, where the interferences between harmonics of modulation fields often disturb the long-term measurements of the sensor. We work out a robust method to eliminate this problem by choosing the modulation frequencies with separations to match the minimum response points of the low-pass filters used in the demodulation processes. We validate the performance of the electronics and the frequency-selection scheme of the modulation fields with corresponding experimental results.

Upregulation of FoxO6 in nucleus pulposus cells promotes DNA damage repair via activation of RAD51.

OBJECTIVE: DNA damage is an essential risk for intervertebral disc degeneration (IDD). Here, we attempted to uncover the effect of FoxO6 and RAD51 on the DNA damage repair of nucleus pulposus (NP) cells in IDD. PATIENTS AND METHODS: We collected the human NP tissues of different degeneration degrees and tested the collagen II, FoxO6, and RAD51 expression. Besides, the IL-1ß induced NP cell model was also used to elucidate the degenerative progress in vitro. We used Chromatin immunoprecipitation (ChIP) and luciferase reporter assay to confirm whether the FoxO6 protein could enhance the RAD51 expression by binding to its promoter. The FoxO6 gene was upregulated in NP cells by vectors transfection. Immunofluorescence staining was used to measure the RAD51 and γH2AX foci formation. Besides, the typical NP cell gene expression was analyzed by RT-PCR. Cell proliferation was determined by CCK-8, and the cell cycle distribution was determined by flow cytometry. RESULTS: Like collagen II, FoxO6 and RAD51 expression were all decreased both in the severe degenerated NP tissue and in the IL-1ß treated NP cells. Upregulation of FoxO6 gene in NP cells enhanced the RAD51 expression via activating the promoter region and inhibited the DNA damage marker γH2AX formation. FoxO6 upregulation alleviated the loss of collagen II, aggrecan, SOD1, and CAT, and suppressed the increase of collagen I/X, TNF-α, and IL-1ß expression, which was affected by IL-1ß. Besides, FoxO6 also helped the proliferation and cell cycle of NP cells with the activation of RAD51. CONCLUSIONS: Upregulation of FoxO6 promotes the DNA repair and maintains the typical phenotype of NP cells, via somehow the mediation of RAD51.

[Experimental study on the effect of three-dimensional porous structures on the vascularization rate of artificial dermis].

Objective: To explore the effects of orienting three-dimensional porous network (type A) and honeycomb briquette-shaped vertically penetrating three-dimensional porous network (type B) on the vascularization rate of artificial dermis. Methods: The experimental research method was used. The artificial dermis was composed of a double layer of silicone layer and scaffold layer. Based on the difference of scaffold layer, they were divided into type A and type B artificial dermis (type A dermis and type B dermis, for short) containing type A and type B structure, respectively. The type A and type B structures were prepared by gradient freeze-drying technique and physical pore-making technique, respectively. The micro-morphology of two kinds of dermis scaffold was observed by scanning electron microscopy. The porosity of two kinds of dermis scaffold was measured by the Pyrex method. According to the method of national medical industry standard, the hydroxyproline content in degradation liquids and their residues in two kind of dermis were determined after degradation at 4, 8, 13, and 24 h, reflecting the degradation rates of two kinds of dermis. According to the random number table, L929 cells were divided into type A dermis group, type B dermis group, negative control group, and positive control group. The positive control group was added with minimum essential medium (MEM) containing 5% dimethyl sulfoxide, The negative control group was added with high-density polyethylene extract, and the other two groups were added with the corresponding extract. At 24 hours after culture, the growth rate of L929 cells was detected by methyl thiazolyl tetrazolium, and the cytotoxicity was graded. L929 cells and human umbilical vein endothelial cells (HUVECs) were inoculated into pore plates with two kinds of dermis preinstalled. On 1, 4, 7, and 14 d after inoculating, the adhesion and growth of L929 cells on the surfaces of the two kinds of scaffolds were detected by immunofluorescence method. On 7 d after inoculating, the migration of the above two kinds of cells into the two kinds of dermal scaffolds was detected by immunofluorescence and hematoxylin-eosin (HE) staining. Three full-thickness skin defect wounds of 5.0 cm×5.0 cm were created on both sides of the back of three 6-month-old healthy male Ba-Ma mini pigs. According to the random number table, six columns of wounds were divided into type A dermis two-step method group, type B dermis two-step method group, and type B dermis one-step method group. The wounds in type A dermis two-step method group and type B dermis two-step method group were transplanted with type A or type B dermis respectively before, and with autologous split-thickness skin grafting later. The wounds in type B dermis one-step method group were transplanted in a synchronous procedure including type B dermis (without silicone layer) and autologous skin grafting simultaneously. The bleeding, exudation, and infection of the wounds on the back in type A dermis two-step method group and type B dermis two-step method group on the 7th day after the second transplantation and in type B dermis one-step method group on the 14th day after the first transplantation were generally observed. The area of autologous skin graft was measured by the transparent film grid method, and the survival rate of autologous skin was calculated. On 4, 7, and 14 d after the first transplantation, the inflammatory cells, fibroblasts (Fbs), and capillary infiltration into the scaffolds of the three groups were detected by HE staining. On 7, 14 d after the first transplantation, the vascularization of the scaffolds was further observed by immunohistochemistry. On 28, 90 d after the first operation, the degradation of the scaffolds of type A dermis and type B dermis was observed by HE staining. Data were statistically analyzed with one-way analysis of variance, independent sample t test, and Bonferroni correction. Results: A large number of round and oval micropores were evenly distributed on the surface of type A scaffold, and the cylindrical hole walls could be observed arranging in a parallel direction in the longitudinal section. The honeycomb briquette-shaped penetrating macropores on the surface of type B scaffold were arranged in an orderly matrix. The pore walls of the honeycomb briquette-shaped penetrating macropores were connected by micropores to form a network structure. The porosity of type A dermis was (93.21±0.72)%, which was similar to (95.88±1.00)% of type B dermis (t=4.653, P>0.05). The degradation rates of type A dermis at 4, 8, 13, and 24 h were similar to those of type B dermis at the corresponding time point (t=0.232, 0.856, 0.258, 7.716, P>0.05). At 24 h after culture, the proliferation rates of L929 cells in the type A dermis group, type B dermis group, and negative control group were significantly higher than those of the positive control group (t=2 393.46, 2 538.27, 1 077.77, P<0.01). The cytotoxicity rating of cells in positive control group was grade 4, while that of the other three groups was grade zero. On 1, 4, 7, and 14 d after inoculation, both L929 cells and HUVECs proliferated in a time-dependent manner in two kinds of dermal scaffolds. The adhesion growth and proliferation rate of the two kinds of cells on the surface of type B dermis was higher than that of type A dermis. On 7 d after inoculation, both L929 cells and HUVECs covered the surface of type B dermis and migrated into one side of the silicone layer. However, the above two kinds of cells migrated slowly into type A dermis, and only a few cells were found on one side of the silicone layer. There was no bleeding, exudation, or infection in the wounds repaired by type A and type B dermis. The survival rate of autologous skin grafting of 6 wounds in each group was 100%. On 4, 7, and 14 d after the first operation, inflammatory cells, Fbs, and capillaries gradually infiltrated into the scaffold layer, and the cell infiltration rate from high to low was type B dermis one-step method group, type B dermis two-step method group, and type A dermis two-step method group. The scaffold in wound in the type B dermis one-step method group gradually collapsed on 28 d after the first operation, and completely degraded in 3 months after the first operation. The scaffold degradation rate of type A dermis two-step method group was similar to that mentioned above. Conclusions: The honeycomb briquette-shaped vertically penetrating three-dimensional porous network structure of type B scaffold can accelerate its vascularization process, which is beneficial to autogenous split-thickness skin in one-step procedure to repair full-thickness skin defects wound in Ba-Ma mini pigs. Compared with the "two-step method" of staged transplantation of type A scaffold and autologous split-thickness skin, and one-step transplantation has equal efficacy and can provide a better choice for wound treatment.

MiR-21 regulates pulmonary hypertension in rats via TGF-ß1/Smad2 signaling pathway.

The article "MiR-21 regulates pulmonary hypertension in rats via TGF-ß1/Smad2 signaling pathway, by F. Ding, T. You, X.-D. Hou, K. Yi, X.-G. Liu, P. Zhang, X.-K. Wang, published in Eur Rev Med Pharmacol Sci 2019; 23 (9): 3984-3992-DOI: 10.26355/eurrev_201905_17828-PMID: 31115027" has been withdrawn from the authors. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17828.