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To explore whether acupuncture can improve sleep disturbance, cognitive impairment and emotional disorders caused by sleep deprivation, and its association with the attenuation of oxidative stress injury in prefrontal cortex. Fifty-two male Sprague-Dawley rats were randomly divided into a control group (n=10), a model group (n=14), a manual acupuncture (MA) group (n=14), and a sham-MA group (n=14). All the groups were established as sleep deprivation models via the modified multiple platform method, except for the control group. Rats in both the MA group and the sham-MA group received corresponding intervention, respectively. After modeling and intervention, the four groups received three behavioral tests, namely sleep monitoring, by comprehensive lab animal monitoring system (CLAMS), Morris water maze (MWM) test and open-field test (OFT), followed by oxygen free radical level test and Western blot (WB) detection for the expression levels of Bax and Bcl-2. The MA group derived more sleep time within 24 h than either the model group or the sham-MA group (both P 0.05), and the escape latency was significantly shorter in these three groups than that in the model group (all P

Background Compared to traditional open surgery, laparoscopic surgery has become a standard approach for colorectal cancer due to its great superiorities including less postoperative pain, a shorter hospital stay, and better quality of life. In 2007, Whiteford et al reported the first natural orifice trans-anal endoscopic surgery (NOTES) sigmoidectomy using transanal endoscopic microsurgery. To date, all cases of NOTES colorectal resection have included a hybrid laparoscopic approach with the use of established rigid platforms. Aim To introduce a novel technique of peroral external traction-assisted transanal NOTES rectosigmoidectomy followed by intracorporeal colorectal end-to-end anastomosis by using only currently available and flexible endoscopic instrumentation in a live porcine model. Methods Three female pigs weighing 25-30 kg underwent NOTES rectosigmoid resection. After preoperative work-up and bowel preparation, general anesthesia combined with endotracheal intubation was achieved. One dual-channel therapeutic endoscope was used. Carbon dioxide insufflation was performed during the operation. The procedure of trans-anal NOTES rectosigmoidectomy included the following eight steps: (1) The rectosigmoid colon was tattooed with India ink by submucosal injection; (2) Creation of gastrostomy by directed submucosal tunneling; (3) Peroral external traction using endoloop ligation; (4) Creation of rectostomy on the anterior rectal wall by directed 3 cm submucosal tunneling; (5) Peroral external traction-assisted dissection of the left side of the colon; (6) Trans-anal rectosigmoid specimen transection, where an anvil was inserted into the proximal segment after purse-string suturing; (7) Intracorporeal colorectal end-to-end anastomosis using a circular stapler by a single stapling technique; and (8) Closure of gastrostomy using endoscopic clips. All animals were euthanized immediately after the procedure, abdominal exploration was performed, and the air-under-water leak test was carried out. Results The procedure was completed in all three animals, with the operation time ranging from 193 min to 259 min. Neither major intraoperative complications nor hemodynamic instability occurred during the operation. The length of the resected specimen ranged from 7 cm to 13 cm. With the assistance of a trans-umbilical rigid grasper, intracorporeal colorectal, tension-free, end-to-end anastomosis was achieved in the three animals. Conclusion Peroral traction-assisted transanal NOTES rectosigmoidectomy followed by intracorporeal colorectal end-to-end anastomosis is technically feasible and reproducible in an animal model and is worthy of further improvements.

Study design Retrospective cohort study. Objective To evaluate the effect of time to first ambulation on recurrence after percutaneous endoscopic lumbar discectomy (PELD). Methods From July 2017 to August 2018, 90 patients with lumbar intervertebral disc herniation underwent PELD surgery. According to the initial walking time, i.e., the time until the patient could walk after the operation, the operations were divided into three groups: early stage, middle stage, and late stage. The follow-up period was 3 months, and complete follow-up data were obtained. The visual analog scale (VAS) and Oswestry disability index (ODI) scores before the operation, at first ambulation, 1 month after the operation, and 3 months after the operation and the recurrence and incidence rates of high magnetic resonance imaging (MRI) signal in the vertebral endplate area were recorded after the operation. Results The success rate was 100% for these 90 cases. The VAS and ODI scores at the first ambulation after the operation significantly improved compared with those before the operation, and the difference was statistically significant. The improvements in the lumbar VAS and ODI scores of the middle- and late-stage groups were better than that of the early-stage group at 1 and 3 months after the operation, and the differences were statistically significant; however, there was no significant difference between the middle- and late-stage groups. The postoperative recurrence rate and rate of high MRI signal in the vertebral endplate area were significantly higher in the early-stage group than in the other two groups, and the difference was statistically significant. Conclusion The time to first ambulation after PELD is an important factor affecting the curative effect of the operation. Early ambulation may be one of the factors affecting recurrence after PELD.

Supplemental material, sj-docx-1-cll-10.1177_09636897221102902 for Clinical Characteristics and Outcome Analysis for HLA Loss Patients Following Partially Mismatched Related Donor Transplantation Using HLA Chimerism for Loss of Heterozygosity Analysis by Next-Generation Sequencing by Andi Wang, Wenjun Li, Fei Zhao, Zhongzheng Zheng, Ting Yang, Sanbin Wang, Jinsong Yan, Jianpin Lan, Shengjin Fan, Mingfeng Zhao, Jianpin Shen, Xin Li, Tonghua Yang, Quanyi Lu, Ying Lu, Hai Bai, Haiyan Zhang, Dali Cai, Ling Wang, Zhiyang Yuan, Erlie Jiang, Fang Zhou and Xianmin Song in Cell Transplantation

In this study, the mechanism of TDP‐43 gene expression on inflammatory factors and Jun N‐terminal kinase (JNK) and p38 mitogen‐activated protein kinase (MAPK) signalling pathways in ischaemic hypoxic stress dependence was investigated. Sixty SD rats were selected and divided into the control group, the osteoarthritis (OA) model group, and the TDP‐43‐mMSCs+OA group. In the OA model group and the TDP‐43‐mMSCs+OA group, OA was established by collagenase injection. Western blotting assays were used to detect the expression of TDP‐43 in cartilage tissues of each rat. The secretion of tumour necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) in the serum of rats was determined by enzyme‐linked immunosorbent assay (ELISA). The formation of cytoplasmic stress granules (SGs) and the expression of receptor for activated c‐kinase 1 (RACK1) were detected by Western blotting assays in each group of rats. The expression of MTK1 and MAPKKK phosphorylation and changes in the JNK and p38 MAPK signalling pathways were detected by Western blotting assays. Compared with the control group, the expression of TDP‐43 in the cartilage tissue of rats in the OA model group was significantly decreased. The expression of TDP‐43 in the cartilage tissue of rats in the TDP‐43‐mMSCs+OA group was significantly higher than that of the control group and the OA model group, which indicates that TDP‐43‐mMSC transplantation was successful. Enzyme‐linked immunosorbent assay results showed that the plasma TNF‐α and IL‐1β levels in the OA model group were significantly increased (P < 0.01) when compared with the control group. However, the secretion of TNF‐α and IL‐1β in the serum of the TDP‐43‐mMSCs+OA group was significantly lower than that of the model group (P < 0.01) but still higher than the control group. This indicates that overexpression of TDP‐43 reduces the inflammatory response induced by OA. Western blotting assays showed that the amount of cytoplasmic SGs in the cartilage tissue of rats in the OA model group was significantly decreased when compared with the control group. The amount of SGs in the cartilage of rats in the TDP‐43‐mMSCs+OA group was significantly higher than that of the model group. The expression of RACK1 in the cartilage tissue of rats in the OA model group was significantly higher than that of the control group. Overexpression of the TDP‐43 gene can interfere with the secretion of inflammatory factors and inhibit the activation of the JNK and p38 MAPK signalling pathways by ischaemic hypoxia stress. Thus, the molecular mechanism of chondrocytopathic lesions was reversed, which provided a new theoretical basis for the treatment of OA.

IntroductionTo investigate the molecular mechanism of icariin (ICA) intervention in TDP-43 mediated chondrocyte lesions of osteoarthritis.Material and methodsHC-α chondrocytes were transfected with TDP-43 lentiviruses to generate TDP-43-overexpressing chondrocytes and treated with 5 μg/mL icariin. The level of TDP-43, JNK, p38 MAPK and relative factors were detected by Western blotting assays. TNF-α and IL-1β in the supernatant were determined by ELISA.ResultsCompared with the HC-α group, TDP-43 expression was significantly increased in the TDP-43-HC-α group and was not significantly different that of the HC-α+ICA group. However, TDP-43 expression in the TDP-43-HC-α+ICA group was significantly lower than that in the TDP-43-HC-α group. ELISA showed that the secretion of TNF-α and IL-1β in the supernatant of the TDP-43-HC-α group was significantly increased (PConclusionsIcariin could interfere with TDP-43-induced secretion of inflammatory factors, inhibit JNK/p38 MAPK signaling. Our findings provided a new theoretical basis for the treatment of osteoarthritis.

Previous studies have suggested that gut microbiota plays a critical role in colorectal cancer (CRC). Although preliminary comparisons of the oral and gut microbiota between CRC and healthy control (HC) patients have been made, the association between microbiome abundance and host clinical factors has not been fully illustrated, especially oral health conditions. Matching samples of unstimulated saliva, cancer tissues or biopsies and stools were collected from 30 CRC and 30 HC patients from Shanghai Jiao Tong University affiliated Renji Hospital for 16S rRNA sequencing analysis. The diversity in salivary and mucosal microbiome, but not stool microbiome of CRC group, was significantly different from that of HC, as demonstrated by the Principal Component Analysis. Logistic regression analysis revealed that older age and higher oral hygiene index (OHI) were independent risk factors for CRC, with odds ratios and 95% confidence intervals of 1.159 (1.045-1.284) and 4.398 (1.328-14.567), respectively. Salivary Firmicutes to Bacteroides ratio in CRC was significantly higher than that in the HC group (P < 0.001), while the mucosal ratio was slightly decreased in CRC (P < 0.05). Salivary Rothia and Streptococcus levels were positively correlated with OHI, while Alloprevotella, Fusobacterium, Peptostreptoccus and Prevotella genera levels were negatively associated with OHI. NetShift analysis revealed that salivary Peptococcus, Centipeda and mucosal Subdoligranulum genus might act as key drivers during the process of carcinogenesis. In conclusion, the current study provides insights into the potential influence of host clinical factors on oral and gut microbiome composition and can be a guide for future studies.

Objective This study evaluated the biomechanical changes in the adjacent vertebrae under a physiological load (500 N) when the clinically relevant amount of bone cement was injected into fractured cadaver vertebral bodies. Methods The embalmed cadaver thoracolumbar specimens in which each vertebral body (T12–L2) had a BMD of < 0.75 g/cm2 were used for the experiment. For establishing a fracture model, the upper one third of the L1 vertebra was performed wedge osteotomy and the superior endplate was kept complete. Stiffness of specimens was measured in different states. Strain of the adjacent vertebral body and intervertebral disc were measured in pre-fracture, post-fracture, and after augmentation by non-contact optical strain measurement system. Results The average amount of bone cement was 4.4 ml (3.8–5.0 ml). The stiffness of after augmentation was significantly higher than the stiffness of post-fracture (p < 0.05), but still lower than pre-fracture stiffness (p < 0.05). After augmentation, the adjacent upper vertebral strain showed no significant difference (p > 0.05) with pre-fracture, while the strain of adjacent lower vertebral body was significantly higher than that before fracture (p < 0.05). In flexion, T12/L1 intervertebral disc strain was significantly greater after augmentation than after the fracture (p < 0.05), but there was no significant difference from that before the fracture (p > 0.05); L1/2 vertebral strain after augmentation was significantly less than that after the fracture (p < 0.05), but there was no significant difference from that before the fracture (p > 0.05). Conclusions PVP may therefore have partially reversed the abnormal strain state of adjacent vertebral bodies which was caused by fracture.

Introduction: GPRC5a plays an important role in many types of cancers with intriguing dual functions. GPRC5a acts as oncogene or tumor suppressor in different types of cancer. It is interesting to illustrate why GPRC5a functions differently.Methods: Data mining method were used to analyze the potential prognostic value of GPRC5a expression for Non-Small Cell Lung Cancer (NSCLC) lung cancer patients. Then we used cell models to further investigate the effect of p53 mutation on GPRC5a expression and the thereafter cell biological behaviors.Results: Our results present here showed High mRNA-level expression of GPRC5a was associated with worse overall survival about lung cancer patients; mutation of p53 gene could result in up regulation of GPRC5a expression and promotion of cell proliferation in lung cancer cells. Our results not only demonstrate the role of GPRC5a as a tumor suppressor in lung cancer, but also revealed the tumor suppressive factor p53 regulated tightly on GPRC5a and cell growth of NSCLC cancer.Conclusions: Our results demonstrated that p53 upregulated GPRC5a expression in NSCLC cells, and the loss of p53 expression in NSCLC may be one of the mechanisms leading to the decreased GPRC5a expression in NSCLC.

Objective To evaluate the clinical effect of the second puncture and injection technique during a percutaneous vertebroplasty (PVP) procedure. Methods Patients treated with a second puncture and injection (group A) or a single puncture and injection (group B) during PVP at our institution during 2010–2017 were reviewed. Vertebral height loss, visual analogue scale (VAS) score, Oswestry disability index (ODI), adjacent vertebral fractures, and cement leakage were compared between the groups. Results A total of 193 patients were enrolled (86 cases in group A, 107 cases in group B). The follow-up period was 15.64 (12–20) months. The loss of anterior (group A 0.01 ± 0.03; group B 0.14 ± 0.17) and middle (group A 0.13 ± 0.12; group B 0.16 ± 0.11) vertebral height in group B was significantly higher than that in group A (P < 0.05). The VAS score and ODI were also significantly higher in group B than in group A at the final follow-up; the VAS score and ODI in group B were 1.65 ± 0.70 and 14.50 ± 4.16, respectively, and those in group A were 1.00 ± 0.74 and 12.81 ± 4.02, respectively (P < 0.05). Three patients in group A and two in group B experienced adjacent vertebral fractures. Regarding mild, moderate, and severe cement leakage, there were 25 (29%), 5 (5%), and 0 cases, respectively, in group A and 28 (26%), 3 (2.8%), and 1 (0.009%) case, respectively, in group B (P > 0.05). Conclusions The second puncture and injection technique may effectively increase the dispersion of cement, thus preventing recompression of the cemented vertebral body, and it does not increase the risk of cement leakage or adjacent vertebral fracture.

To evaluate the clinical effect of ultra-early injection (before the phase of "tooth-paste-like") of low-viscosity cement in percutaneous vertebroplasty (PVP) for treating osteoporotic vertebral compression fractures (OVCFs).Two hundred sixty-one patients who had PVP procedures with low-viscosity cement (ultra-early injection: 145, normal injection: 135) were included from July 2010 to July 2016 in our hospital. Visual Analog Scale (VAS), Oswestry Disability Index (ODI), Cobb angle, cement leakage, and adjacent vertebral fractures were evaluated. The follow-up period was over 12 months.VAS 3.0 d after surgery was significantly reduced in the ultra-early injection group compared to that in the control group (P = 0.00), but no difference was found at the final follow-up (P = 0.53). Similar results were found for ODI. The Cobb angle in both groups was recovered after PVP (P 0.05); however, in the control group, the Cobb angle at the final follow-up was significantly increased compared with that 3.0 d after surgery (P = 0.00). There was a significant difference in the Cobb angle between the two groups at the final follow-up (P = 0.00). Regarding cement leakage, there were no significant differences in terms of mild (P = 0.58), moderate (P = 0.68), or severe leakage (P = 0.52). Seven patients in the control group had adjacent vertebral fractures, but only one patient in the ultra-early injection group experienced adjacent fractures (P = 0.03).Ultra-early injection of low-viscosity cement during PVP procedures in the treatment of OVCFs not only quickly and significantly relieves pain, reduces the incidence of adjacent vertebral fractures, and prevents progressive kyphotic deformity, but also does not increase the risk of cement leakage when compared with that of the traditional injection procedure.

Objective This study mainly aimed to evaluate complications of cement leakage for osteoporotic thoracolumbar vertebral compression fractures by PVP using HVC, and access the clinical efficacy. Methods Between May 2013 and June 2015, 66 patients with osteoporotic thoracolumbar vertebral compression fractures, who underwent PVP (36 HVC and 30 LVC) in our hospital, were enrolled. Cement leakage, Visual Analog Scale (VAS), Oswestry Disability Index (ODI), refracture of the cemented vertebrae, and adjacent vertebral fractures were evaluated. The follow-up time was 1 year. Results The overall cement leakage rate was 30.55% in the HVC group, lower than 77.77% obtained in the LVC group (P = 0.00). The incidence rates of cement leakage into paravertebral area (P = 0.02) and vein (P = 0.04) in the HVC group were significantly lower than those of the LVC group; however, no differences were found for disc space (P = 0.72) and intraspinal space (P = 0.58). There were no differences in VAS, ODI, refracture of cemented vertebrae, and adjacent vertebral fracture between the two groups (P > 0.05). Conclusions PVP using HVC not only can reduce cement leakage, especially in the paravertebral area and peripheral vein, but also has satisfactory clinical effect.

BACKGROUND It is unclear whether hepatitis B virus (HBV) itself causes iron metabolism disorder in patients with chronic hepatitis B (CHB). In this study, we investigated the effect of HBV on iron metabolism at the clinical and cellular levels to determine the pathogenesis of CHB. MATERIALS We enrolled 41 CHB patients and 20 healthy controls (HCs) in a retrospective study. Parameters of iron status included serum iron (SI), ferritin (SF), transferrin (TRF), soluble transferrin receptor (sTfR), transferrin saturation (TS), total iron-binding capacity (TIBC), unsaturated iron-binding capacity (UIBC), and hepcidin. Liver function indicators included serum alanine transaminase (ALT) and albumin. Furthermore, we investigated the correlations between iron markers and liver function indicators. Finally, the alterations in SF, TRF, transferrin receptor (TfR), and hepcidin expression were detected by RT-PCR, western blot, and cell immunofluorescence after HepG2 cells and Huh7 cells were transfected with the pSM2-HBV plasmid. We also measured these alterations between HepG2 cells and HepG2.215 cells. The significance of differences was analyzed by SPSS version 17.0. RESULTS Compared with healthy controls, the CHB patients were more likely to have lower levels of serum hepcidin, TRF, sTfR, TIBC, and UIBC and higher levels of SI, SF, and TS (p < 0.05, all). In CHB patients, the levels of SI and SF correlated positively with ALT concentrations, and the serum hepcidin concentrations correlated positively with albumin concentrations (p < 0.05, all). The expression levels of ferritin, transferrin, and hepcidin mRNA and protein were significantly higher in HepG2.215 cells than in HepG2 cells, while expression levels of TfR were lower. The alterations in these iron markers in HepG2 and Huh7 cells that were transfected with pSM2-HBV plasmid were consistent with those in HepG2.215 cells. CONCLUSIONS Serum iron markers tended to be abnormal in CHB patients. In hepatocytes, HBV promoted the expression of ferritin, transferrin, and hepcidin, while it inhibited the expression of TfR.

The accurate diagnosis of sporotrichosis and identification at the species level are critical for public health and appropriate patient management. Compared with morphological identification methods, molecular diagnostic tests are rapid and have high sensitivity and standardized operating processes. Therefore, we designed a novel multiplex real-time polymerase chain reaction (PCR) method based on the calmodulin (CAL) gene for the identification of clinically relevant Sporothrix species: S. globosa, S. schenckii s. str., and S. brasiliensis. We evaluated the assay with clinical and spiked samples and assessed its diagnostic performance by comparing the results to those of culture and species-specific PCR. Thirty-three DNA templates were used to detect assay specificity, and three plasmids were constructed to create a standard curve and determine the limits of detection (LODs). For nucleic acid detection, the sensitivity and specificity reached 100%. The LODs were 10 copies, 10 copies and 100 copies for S. globosa, S. schenckii s. str and S. brasiliensis, respectively. For the clinical samples, the positive detection rates by culture, species-specific PCR and the multiplex real-time PCR assay were 87.9% (29/33), 39.4% (13/33), and 93.9% (31/33), respectively. For the spiked samples, the positive detection rates were both 100% for S. schenckii s. str and S. brasiliensis. Based on the above results, compared with culture and other molecular diagnosis methods, the novel multiplex real-time PCR assay is effective, fast, accurate, and highly sensitive. It has a lower reaction cost and lower sample volume requirements, can detect co-infections, and allows for standardized operation and easier interpretation of results. In the future, this assay could be developed into a commercial kit for the diagnosis and identification of S. globosa, S. schenckii s. str, and S. brasiliensis., Author summary Sporotrichosis is a subacute or chronic infectious disease caused by dimorphic fungi of Sporothrix spp. The genus Sporothrix consists of several species with different geographic distributions, virulence, and antifungal susceptibilities, making species-level identification necessary. S. brasiliensis, S. globosa, S. schenckii s. str and S. luriei make up the “pathogenic clade” of the genus Sporothrix. Importantly, S. luriei has a low clinical-epidemiological impact within this genus. Therefore, we designed a novel multiplex real-time PCR method using fluorescent probes for the identification of S. globosa, S. schenckii s. str, and S. brasiliensis. We designed a pair of primers based on the conserved sequence of the calmodulin gene of Sporothrix spp. and probes with different fluorescent signals based on the divergent sequences of S. globosa, S. schenckii s. str and S. brasiliensis. Through the verification of nucleic acid, clinical and spiked sample detection, the multiplex real-time PCR could quickly and accurately identify the three clinically relevant species of Sporothrix spp. with high sensitivity. This new assay could be applied in epidemiology, clinical diagnosis, and experiments with sporotrichosis to control new outbreaks, reduce diagnostic and identification time, and improve test efficiency.

Objective: To systematically evaluate the efficacy and diagnostic value of epicardial adipose tissue in coronary artery disease. Methods: The authors searched PubMed, Embase, Web of Science, CNKI and CBM from the database to January 2019, and collected articles on the clinical trials of epicardial adipose tissue in coronary heart disease. The improved Jadad scale was used for quality evaluation, and Revman 5.3 software was used for metaanalysis of the included literature. Results: Finally, 21 articles were included, including a total of 4049 patients. EAT was 12.21 2.62 mm in the CAD group and 9.92 1.37 mm in the non-CAD group. EAT was 2.22 1.86 mm in the CAD group and 1.8 1.4 mm in the non-CAD group. The random effect model was adopted for the study (I2=95%, p

To investigate the characteristics of the sandstones under two fracture modes which are tensile fracture and shear fracture, we proposed three-point bending test and a modified shear test. Meanwhile, numerical simulations using particle flow code (PFC) and acoustic emission (AE) analysis are performed to obtain their differences. The AE hits and amplitude analyses prove that no matter which mode, they experience obvious three stages. The AE hit curve and amplitude are lower in the first stage. Then, the curve goes up steadily and the amplitude value becomes relatively large in the second stage. Entering into the third stage, the curve increases quickly to its peak value and a lot of AE signals with higher amplitude are generated. Moreover, AE b-value analysis is also used to make crack magnitude assessment. It was obvious that the distribution of b-value curve can be divided into three stages, which are just corresponding to the division of AE hits and amplitude distributions. The b-values are diverse and fluctuating largely in stage I and then show small fluctuations and have increasing trends in the whole process in stage II whereas finally exhibit sharp down to the minimum points at the stage III. The average b-values for tensile fracture tests are in the range of 0.877–1.09 whereas for shear fracture tests are within the range of 0.815–0.876. Comparing to tensile fracture, when shear fracture occurs in the rock sample, the AE energy releases faster and the proportion of large magnitude cracks is larger. The research results in this article could make more instructive and applied value to in situ non-destructive monitoring of fissure development for engineering rock mass.

Stem cell therapy is widely employed in treating osteoarthritis (OA), and bone marrow-derived mesenchymal stem cells (BMSCs) has gradually become the most attractive new method for treating OA due to the benefit for cartilage tissue repair. However, the apoptosis in the neural stem cell transplantation severely decreases repairing efficacy. Icariin has been reported to exert multiple effects on BMSCs, including its proliferation, osteogenic, and chondrogenic differentiation. However, its effects on the injury induced by oxygen, glucose and serum deprivation (OGD) remains unknown. We prospectively investigated the role of ICA on rabbit BMSCs under conditions of OGD. Firstly, BMSCs were cultured under conditions of OGD, ICA relieved OGD-induced cell damage by promoting cell proliferation and suppressing apoptosis. Secondly, Markers of endoplasmic reticulum stress (ERs), ER stress IRE-1 pathway, and autophagy were both inhibited by ICA via inhibition of phosphor-extracellular regulated protein kinases (p-ERKs), p-P38, p-c-Jun N-terminal kinase (p-JNK) or si-MAPK. Finally, decrease of ERs marker levels enhanced protective effect of ICA against OGD-induced injury by limiting apoptosis and autophagy. Moreover, an autophagy inhibitor (3-methyladenine: 3-MA) contributed to a synergistic effect in conjunction with ICA, in promoting cell proliferation, suggesting that ICA exerts anti-ERs and anti-autophagy effects in OGD-treated BMSCs. Therefore, ICA protected rabbit BMSCs from OGD-induced apoptosis through inhibitory regulation of ERs-mediated autophagy related to the MAPK signaling pathway, which provided insights for a potential therapeutic strategy in OA.

The molecular mechanisms that control the development of colorectal cancer (CRC) remain poorly defined. Here we show Synbindin promoted CRC oncogenesis by activating Wnt signaling and altering gut microbiome. Synbindin upregulation in human CRCs was associated with poor patient prognosis. Intestine-specific disruption of Synbindin balanced the disturbed gut microbiota and protected mice against tumor formation in the colitis-associated cancer (CAC) model. The protective role was compromised after gut microbiota depletion. In host, increased goblet cells and mucin2 expression, together with increased intestinal epithelial cells (IECs) apoptosis and decreased epithelial proliferation were observed. Further transcriptomic sequencing identified Wnt signaling a major regulatory node downstream of Synbindin. Combined molecular and cellular characterizations revealed that Synbindin confers Disheveled-3 (DVL3)-based signalosome assembly and acts as a modular scaffold for DVL3 and Axin2 complex, orchestrating the intensity of Wnt signaling. These findings identify a critical role of Synbindin in gut microbiome composition and Wnt signaling activation in colorectal carcinogenesis, and highlight Synbindin as an adaptor protein with multifaceted roles.

Bone marrow-derived mesenchymal stem cells (BMSCs) are widely used in tissue engineering for regenerative medicine due to their multipotent differentiation potential. However, their poor migration ability limits repair effects. Icariin (ICA), a major component of the Chinese medical herb Herba Epimedii, has been reported to accelerate the proliferation, osteogenic, and chondrogenic differentiation of BMSCs. However, it remains unknown whether ICA can enhance BMSC migration, and the possible underlying mechanisms need to be elucidated. In this study, we found that ICA significantly increased the migration capacity of BMSCs, with an optimal concentration of 1 μmol/L. Moreover, we found that ICA stimulated actin stress fiber formation in BMSCs. Our work revealed that activation of the MAPK signaling pathway was required for ICA-induced migration and actin stress fiber formation. In vivo, ICA promoted the recruitment of BMSCs to the cartilage defect region. Taken together, these results show that ICA promotes BMSC migration in vivo and in vitro by inducing actin stress fiber formation via the MAPK signaling pathway. Thus, combined administration of ICA with BMSCs has great potential in cartilage defect therapy.

Platelet-rich plasma (PRP) has been reported to alleviate degenerative pathological damage to joint cartilage. This study aimed to investigate the effect of PRP on Wnt/β-catenin signaling in rabbit chondrocytes.Using 3-month-old New Zealand white rabbits, PRP was prepared from venous blood, and chondrocytes were cultured from knee joint cartilage and identified by staining for type II collagen and proteoglycan. The effects of PRP on chondrocyte viability were measured. The chondrocytes were divided into 5 groups: control, IL-1β, PRP (100-fold dilution), Dkk-1 (100ng/mL) and Dkk-1+PRP. The IL-1β, PRP, Dkk-1 and Dkk-1+PRP groups were treated with interleukin (IL)-1β (50μL, 10μg/mL) for24h. Chondrocyte morphology was observed by electron microscopy. Levels of carboxy terminal peptide (CTX-II) and cartilage oligomeric matrix protein (COMP) in culture media were measured by ELISA. Wnt-1, β-catenin and GSK-3β mRNA and protein expression were determined by RT-PCR and western blot respectively.PRP enhanced chondrocyte proliferation. Chondrocytes in the IL-1β group showed ultrastructural abnormalities that were less pronounced in the PRP, Dkk-1 and Dkk-1+PRP groups. CTX-II and COMP concentrations were higher in the IL-1β group than in the control, PRP, Dkk-1 and Dkk-1+PRP groups (P0.05). The IL-1β group had higher mRNA and protein Wnt1 and β-catenin levels and lower GSK-3β levels than the control, PRP, Dkk-1 and Dkk-1+PRP groups (P0.05).PRP may protect chondrocytes activated by IL-1β via inhibiting Wnt/β-catenin signaling.

Cardiomyocyte apoptosis is closely associated with the pathogenesis of heart failure. Jujuboside A (JUA) is a type of saponin isolated from the seeds of Zizyphus jujuba. In traditional Chinese medicine, it is believed that JUA possesses multiple biological effects, including antianxiety, antioxidant and anti‑inflammatory activities. The present study aimed to evaluate the effects of JUA on norepinephrine (NE)‑induced apoptosis of H9c2 cells and to investigate its underlying mechanisms. Rat H9c2 cardiomyocytes were pretreated with JUA and were then exposed to NE as an in vitro model of myocardial apoptosis. A cell viability assay, scanning electron microscopy, transmission electron microscopy, flow cytometry assay, acridine orange/ethidium bromide staining, reverse transcription‑quantitative polymerase chain reaction and western blotting, all revealed that NE induced H9c2 cell apoptosis. The results demonstrated that NE inhibited cell viability, and enhanced cell damage and apoptosis of H9c2 cells. Conversely, pretreatment with JUA was able to reverse NE‑induced decreased cell viability and increased apoptosis. Furthermore, JUA suppressed upregulation of the B‑cell lymphoma 2 (Bcl‑2)‑associated X protein/Bcl‑2 ratio, and inhibited the increased protein expression levels of cleaved caspase‑3 and cleaved caspase‑9 following NE exposure. However, the protein expression levels of cleaved caspase‑12 and cleaved caspase‑8 were not significantly altered following exposure to NE or JUA pretreatment. In addition, in JUA‑pretreated cells, the protein expression levels of phosphorylated (p)‑p38 and p‑c‑Jun N‑terminal kinase were downregulated compared with in NE‑treated cells. Furthermore, JUA regulated the activation of extracellular signal‑regulated kinase (ERK) in NE‑treated cells and significantly increased the expression levels of p‑AKT. Taken together, these data suggested that JUA may protect against NE‑induced apoptosis of cardiomyocytes via modulation of the mitogen‑activated protein kinase and AKT signaling pathways. Therefore, JUA may be considered a potential therapeutic strategy for the treatment of heart disease.

Background Lumbar disc disease has a disabling impact on global people with heavy burden on society, mainly consisting of lumbar disc degeneration (LDD) and lumbar disc herniation (LDH). The recently released lumbar disc nomenclature version 2.0 deepens our understandings on the diseases. Consequently, there is an urgent need to clarify the occurrence and distribution features of LDD and LDH in a large-scale sample in terms of the novel version. Question/purposes We asked: (1) Is there a difference in the occurrence and distribution hallmarks of LDD and LDH in a population-based large-scale sample? (2) Does the novel nomenclature version bring novel vision on lumbar disc disease? Methods Five thousand two hundred eighty-eight consecutive cases (26,440 lumbar discs) undergoing lumbar spine MRI were retrospectively included from Jan 2008 to Dec 2010 in a territory university hospital. Five hundred nine cases were excluded. There were 2727 males (51.57%) and 2561 females (48.43%) with a mean age of 43.73 years. Both T1 and T2 weighted lumbar MRI images from L1/2 to L5/S1 were profoundly analyzed in axial and sagittal planes. We classified lumbar discs in terms of version 2.0. Results The occurrence of LDH and LDD was 14.18 and 44.23% in average, respectively. Notably, lumbar spine discs were more prone to LDD than LDH. L4/5 was the most frequent level in terms of LDH (26.08%) and LDD (56.09%), followed by L5/S1 (LDH: 24.09%; LDD: 55.33%), then L3/4, L2/3 and L1/2 in ranking order. The prevalence of LDH and LDD in upper lumbar discs from L1/2 to L3/4 was significant lower than the average prevalence rate (P

Background This study focused on the mechanisms where icariin inhibited chondrocyte apoptosis and angiogenesis by regulating the TDP‐43 signaling pathway. Methods A rat osteoarthritis (OA) model was established by collagenase injection. Histological examination of the articular cartilage and synovial tissue was performed 6 weeks after operation. Cartilage cell line overexpressing TDP‐43 and mesenchymal stem cell line (TDP43‐MSCs) of the rat TDP43 gene were established. Results In OA rats transplanted with TDP43‐mMSCs, TDP43 was highly expressed in chondrocytes (TDP43‐HC), while TDP43 expression was low in HC and MSCs‐HC (p

The conventional valves used in the choke manifold for well control usually have intensely nonlinear characteristic of the match between pressure choking and vale opening. Therefore, it is difficult to achieve the precise choking pressure that is essential to the managed pressure drilling (MPD) through a single throttle within the scope of full opening. However, the parallel choke manifold proposed in this paper, i.e. a program of parallel throttling, can enhance the accurate choking wellhead backpressure for MPD. The fine linear choking principle and feasibility of the parallel choke manifold were analyzed in detail by calculating the resistance modulus of bypass valves. The formula calculated the resistance modulus of each valve under different working state. Furthermore, the laboratory tests shown the parallel throttle method can achieve the control of fine choking pressure by switching one or more valves in the branch pipes. Therefore, the proposed parallel choke manifold system can reduce the requirements for the linear capability of valves and provide some references to further study the surface fine choking pressure system for MPD.

Objective To observe the effects of oridonin on proliferation and apoptosis of myeloma RPMI8226 cells and to investigate the potential underlying mechanisms. Methods RPMI8226 cells were treated with various concentrations of oridonin. Cell proliferation was analyzed using the thiazolyl blue tetrazolium bromide method. Ultramicrostructure was observed by transmission electron microscopy. Annexin-V/PI staining and flow cytometry was performed to determine cell apoptosis. Expression of apoptosis-related proteins was evaluated by western blot analysis. Results Oridonin suppressed the proliferation of RPMI8226 cells and induced apoptosis in a time- and dose-dependent manner. Transmission electron microscopy confirmed apoptotic morphology upon treatment with 20 µmol/L oridonin and western blot revealed decreased expressions of the apoptosis suppressors survivin, Bcl-2 and pro-caspase-3 proteins, and the increased expression of the apoptosis inducer Bax. Conclusion Our results show that oridonin exhibits an inhibitory effect on the proliferation of RPMI8226 cells and induces apoptosis. This is associated with altering the balance between Bcl-2 and Bax protein expressions and decreased survivin and pro-caspase-3 expressions.

Vulcanization and reinforcement are two important factors contributing to the properties of vulcanized rubber. In order to investigate the influence of carbon black (CB) on chemical crosslinking, three groups of samples with different crosslink densities were prepared. In each group with the same crosslink density, different amounts of CB were introduced. Data fitting showed that delta torque (ΔM = M H–M L, the difference between the highest and lowest torques during curing) in the cure curves of each group had a good linear relationship with CB load and extrapolation of the fitting lines almost intercepted the x coordinate at the same value, which indicated that CB had no influence on the chemical crosslinking of the rubber. To verify the above result, a series of nonfilled natural rubber (NR) vulcanizates with different crosslink densities were studied using equilibrium swelling and the swelling ratios were compared with those of corresponding CB filled rubbers with the same sulfur and accelerator amoun...

Study design Vertebral endplate tissues were obtained from patients with degenerated lumbar disc classified as Modic type 1 or type 2 and investigated with immunohistochemical staining and the DNA nick end labelling techniques. Objective To examine the expression of fas receptor (FasR) and apoptosis in the endplate cells isolated from patients with degenerative disc disease and to see whether they are associated with the pathological change of endplate. Methods Fifty-six degenerated lumbar endplate specimens were obtained from the patients with degenerative disc disease categorized as type Modic I or II in magnetic resonance imaging (MRI) and eight nondegenerated specimens as control (vertebra burst fracture patients without degenerative change in MRI) during surgical procedures. Immunohistochemical staining was performed to determine the presence of FasR and apoptosis in sections of those endplate tissues. To investigate whether the FasR expression and apoptosis in endplate were influenced by degeneration and ageing of the discs, the level of FasR expression and apoptosis in the changed and unchanged endplates was analysed. Results FasR were expressed in the cytoplasm of the endplate cells. A higher degree of FasR expression and apoptosis in endplate cells in degenerated discs was found than that in nondegenerated discs. In cell culture, the level of FasR expression and apoptosis in cells from the degenerative endplates was higher than those in unchanged endplates. The percentage of FasR-positive endplate and apoptotic endplate cells correlated significantly with the patient's age ( r = 0.301, p r = 0.307, p r = 0.047, p > 0.05; r = 0.066, p > 0.05, respectively). Conclusion This is the first study to compare the expression of FasR and apoptosis on vertebral endplate cells in degenerated disc with in nondegenerated disc. The results show that the endplate in degenerated disc may undergo fas-mediated apoptosis and vice versa, endplate degenerative changes may promotes apoptosis of the endplate cells within degenerated disc.

Background:High-viscosity cement (HVC) has been gradually applied in percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP). Although HVC has been reported to reduce cement leakage, different opinions exist. To assess the complications of HVC in cement leakage in the treatment

The design and fabrication of new type viscosity sensor on molecular scale will enrich the detection means in both physical and life sciences. In this work, a rotor-containing fluorescent dye was prepared, which can reveal a typical viscosity-sensitive behavior due to the environment-induced non-radiative decay. The viscosity-sensitive behavior of this dye could be tunable (locked and activated) in aqueous environment, either with the reversible light-driven trans / cis -photoisomerization of the cyanostilbene moiety, or with the reversible assembly/disassembly process of β-cyclodextrin (β-CD) to the rotor part. Such dual-mode tunable viscosity sensitivity can be well distinguished by fluorescent spectra. A slope method, on double-logarithmic plots of the fluorescent signal to the corresponding environmental viscosity, makes a quantitative estimation to the tunable viscosity sensitivity, featuring the ‘Activated’ and the ‘Locked’ state in this molecular-scale viscometer.

Aims: To investigate the antitumor effects of probiotics Clostridium butyricum and Bacillus subtilis on colorectal cancer (CRC) progression. Materials & methods: The effects of C. butyricum and B. subtilis on CRC cells were studied. Male C57BL/6 mice with 1,2-dimethylhydrazine dihydrochloride (DMH)-induced CRC were intervened by these two probiotics and the antitumor effects were examined by comparing the tumor incidence and detecting the inflammatory and immune-related markers. Results & conclusions: C. butyricum and B. subtilis inhibited the proliferation of CRC cells, caused cell cycle arrest and promoted apoptosis. In vivo, these two probiotics inhibited the development of DMH-induced CRC. The molecular mechanism involved reduced inflammation and improved immune homeostasis. This work establishes a basis for the protective role of probiotics B. subtilis and C. butyricum in intestinal tumorigenesis.

AIM: To demonstrate the feasibility and reproducibility of a pure natural orifice transluminal endoscopic surgery (NOTES) gastrojejunostomy using holing followed by interrupted suture technique using a single endoloop matched with a pair of clips in a non-survival porcine model. METHODS: NOTES gastrojejunostomy was performed on three female domestic pigs as follows: Gastrostomy, selection and retrieval of a free-floating loop of the small bowel into the stomach pouch, hold and exposure of the loop in the gastric cavity using a submucosal inflation technique, execution of a gastro-jejunal mucosal-seromuscular layer approximation using holing followed by interrupted suture technique with endoloop/clips, and full-thickness incision of the loop with a Dual knife. RESULTS: Pure NOTES side-to-side gastrojejunostomy was successfully performed in all three animals. No leakage was identified via methylene blue evaluation following surgery. CONCLUSION: This novel technique for preforming a gastrointestinal anastomosis exclusively by NOTES is technically feasible and reproducible in an animal model but warrants further improvement.

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder that is thought to be caused in part by the age-related accumulation of amyloid-β (Aβ) in the brain. Recent findings have revealed that nitric oxide (NO) modulates the processing of amyloid-β precursor protein (APP) and alters Aβ production; however, the previously presented data are contradictory and the underlying molecular mechanisms are still incomplete. Here, using human SH-SY5Y neuroblastoma cells stably transfected with wild-type APPwt695, we found that NO, derived from NO donor sodium nitroprusside (SNP), bi-directionally modulates APP processing in vitro. The data from ELISA and Western blot (WB) tests indicated that SNP at lower concentrations (0.01 and 0.1 μM) inhibits BACE1 expression, thus consequently suppresses APP β-cleavage and decreases Aβ production. In contrast, SNP at higher concentrations (10 and 20 μM) biases the APP processing toward the amyloidogenic pathway as evidenced by an increased BACE1 but a decreased ADAM10 expression, together with an elevated Aβ secretion. This bi-directional modulating activity of SNP on APP processing was completely blocked by specific NO scavenger c-PTIO, indicating NO-dependent mechanisms. Moreover, the anti-amyloidogenic activity of SNP is sGC/cGMP/PKG-dependent as evidenced by its reversal by sGC/PKG inhibitions, whereas the amyloidogenic activity of SNP is peroxynitrite-related and can be reversed by peroxynitrite scavenger uric acid. In summary, these present findings predict a double-edged role of NO in APP processing in vitro. Low (physiological) levels of NO inhibit the amyloidogenic processing of APP, whereas extra-high (pathological) concentrations of NO favor the amyloidogenic pathway of APP processing. This preliminary study may provide further evidence to clarify the molecular roles of NO and NO-related signaling in AD and supply potential molecular targets for AD treatment.

Both the Global Registry of Acute Coronary Events (GRACE) risk score and neutrophil count could predict clinical outcomes in patients with acute coronary syndromes. However, the ability of them to identify high risk patients leaves room for improvement. The purpose of the present study was to evaluate whether the combination of them could have a better performance in predicting clinical outcomes in patients with ST-elevation myocardial infarction (STEMI).A total of 1287 consecutive STEMI patients were recruited at two centers in China. Neutrophil count was measured and the GRACE risk score was calculated.During a median period of 37 months (IQR, 29-47), 135 (10.9%) patients had major adverse cardiovascular events (MACE), including 116 all-cause death. Neutrophil count and the GRACE risk score were higher in patients with MACE. Both neutrophil count and the GRACE score were significant and independent predictors for MACE [HR: 1.260 (1.203-1.319), P 0.001; HR: 1.007 (1.002-1.011), P 0.001; respectively). Combination of them increased the area under the ROC (0.698 vs. 0.796, P 0.001). The addition of neutrophil count to GRACE model enhanced net reclassification improvement (0.637, P = 0.020) and integrated discrimination improvement (0.180, P 0.001), suggesting effective discrimination and reclassification.Both neutrophil count and the GRACE risk score are independent predictors for MACE in patients with STEMI. A combination of them could derive a more accurate prediction for clinical outcomes in these patients.

GP41, a major glycoprotein, identified in the occlusion-derived virions (ODV) of baculoviruses, is required for the egress of nucleocapsids from the nucleus in the pathway of budded virion (BV) synthesis. Using the polymerase chain reaction (PCR), the open reading frame (ORF) ofSpodoptera lituranucleopolyhedrovirus (SpltMNPV)gp41gene was obtained from SpltMNPV genomic DNA. The PCR product was cloned into pMD18-T vector to get the recombinant plasmid (pT-gp41). Thegp41gene was recombinedin vitrowith prokaryotic expression vector pQE30 and transformed intoEscherichia coliM15 [pREP4]. The M15 [pREP4] strain, containinggp41recombinant plasmid, expressed a 37.9 kDa 6×His-tag fusion protein after induction with 1 mmol/l isopropylthio-β-d-galactoside (IPTG). The fusion protein was purified with a nickel-nitrilotriacetic acid (Ni–NTA) resin column and used as the immunogen to raise GP41-specific antibody. Western blotting analysis indicated that the antibody was suitable to be used for further analysis of GP41 protein.

One pregnant captive Irrawaddy dolphin (Orcaella brevirostris), with a body length of 225 cm, was found dead on 8 June 2009. The dolphin was anorexic and circling at the bottom of the pool before death. Laboratory tests revealed an increased leukocyte count and decreased platelet count; increased erythrocyte sedimentation rate; and slightly decreased red blood cell count, hemoglobin, and hematocrit. Alkaline phosphatase, creatinine, and glucose were significantly decreased. Moreover, uric acid and alanine aminotransferase and aspartate aminotransferase levels were elevated. A 57-cm fetus was recovered. The respiratory system, intestinal mucosa, mesentery and mesenteric lymph nodes, and spleen were congested and hemorrhagic. The heart, liver, and kidney appeared normal. Klebsiella spp. and Staphylococcus aureus were identified in the amniotic fluid. This is the first case report of bacterial infection in an Irrawaddy dolphin.

BACKGROUND Current studies evaluating the association of tea consumption and bone mineral density (BMD) have yielded inconsistent findings. Therefore, we conducted a meta-analysis to assess the relationship between tea consumption and BMD. METHODS The PubMed, Embase, and Cochrane Library databases were comprehensively searched, and a meta-analysis performed of all observational studies assessing the association of tea consumption and BMD. Forest plots were used to illustrate the results graphically. The Q-test and I statistic were employed to evaluate between-study heterogeneity. Potential publication bias was assessed by the funnel plot. RESULTS Four cohort, 1 case-control, and 8 cross-sectional studies including a total of 12,635 cases were included. Tea consumption was shown to prevent bone loss [odds ratio (OR): 0.66; 95% confidence interval (CI), 0.47-0.94; P = 0.02], yielding higher mineral densities in several bones, including the lumbar spine [standardized mean difference (SMD): 0.19; 95% CI, 0.08-0.31; P = 0.001], hip (SMD: 0.19; 95% CI, 0.05-0.34; P = 0.01), femoral neck [mean difference (MD): 0.01; 95% CI, 0.00-0.02; P = 0.04], Ward triangle (MD: 0.02; 95% CI, 0.01-0.04; P = 0.001), and greater trochanter (MD: 0.03; 95% CI, 0.02-0.04; P

Purpose CO2 leakage along the trocar (chimney effect) has been proposed to be an important factor underlying port-site metastasis after laparoscopic surgery. This study aimed to test this hypothesis by comparing the incidence of port-site metastasis between B-ultrasound-guided and laparoscopically-assisted hyperthermic intraperitoneal perfusion chemotherapy (HIPPC). Materials and Methods Sixty-two patients with malignant ascites induced by gastrointestinal or ovarian cancer were divided into two groups to receive either B-ultrasound-guided or laparoscopically-assisted HIPPC. Clinical efficacy was assessed from the objective remission rate (ORR), the Karnofsky Performance Status (KPS) score, and overall survival. The incidence of port-site metastasis was compared between the two groups. Results Patients in the B-ultrasound (n=32) and laparoscopy (n=30) groups were comparable in terms of age, sex, primary disease type, volume of ascites, and free cancer cell (FCC)-positive ascites. After HIPPC, there were no significant differences between the B-ultrasound and laparoscopy groups in the KPS score change, ORR, and median survival time. The incidence of port-site metastasis after HIPPC was not significantly different between the B-ultrasound (3 of 32, 9.36%) and laparoscopy (3 of 30, 10%) groups, but significantly different among pancreatic, gastric, ovarian, and colorectal cancer (33.33, 15.79, 10.00, and 0.00%, p

Mutations in the SCN1A gene-encoding voltage-gated sodium channel α-I subunit (Nav1.1) cause various spectrum of epilepsies including Dravet syndrome (DS), a severe and intractable form. A large number of SCN1A mutations identified from the DS patients lead to the loss of function or truncation of Nav1.1 that result in a haploinsufficiency effects, indicating that the exact expression level of SCN1A should be essential to maintain normal brain function. In this study, we have identified five variants c.*1025T>C, c.*1031A>T, c.*1739C>T, c.*1794C>T and c.*1961C>T in the SCN1A 3' UTR in the patients with DS. The c.*1025T>C, c.*1031A>T and c.*1794C>T are conserved among different species. Of all the five variants, only c.*1794C>T is a novel variant and alters the predicted secondary structure of the 3' UTR. We also show that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) only binds to the 3' UTR sequence containing the mutation allele 1794U but not the wild-type allele 1794C, indicating that the mutation allele forms a new GAPDH-binding site. Functional analyses show that the variant negatively regulates the reporter gene expression by affecting the mRNA stability that is mediated by GAPDH's binding, and this phenomenon could be reversed by shRNA-induced GAPDH knockdown. These findings suggest that GAPDH and the 3'-UTR variant are involved in regulating SCN1A expression at post-transcriptional level, which may provide an important clue for further investigating on the relationship between 3'-UTR variants and SCN1A-related diseases.

Aim To introduce natural orifice transgastric endoscopic surgery (NOTES) tube ileostomy using pelvis-directed submucosal tunneling endoscopic gastrostomy and endoscopic tube ileostomy. Methods Six live pigs (three each in the non-survival and survival groups) were used. A double-channeled therapeutic endoscope was introduced perorally into the stomach. A gastrostomy was made using a 2-cm transversal mucosal incision following the creation of a 5-cm longitudinal pelvis-directed submucosal tunnel. The pneumoperitoneum was established via the endoscope. In the initial three operations of the series, a laparoscope was transumbilically inserted for guiding the tunnel direction, intraperitoneal spatial orientation and distal ileum identification. Endoscopic tube ileostomy was conducted by adopting an introducer method and using a Percutaneous Endoscopic Gastrostomy Catheter Kit equipped with the Loop Fixture. The distal tip of the 15 Fr catheter was placed toward the proximal limb of the ileum to optimize intestinal content drainage. Finally, the tunnel entrance of the gastrostomy was closed using nylon endoloops with the aid of a twin grasper. The gross and histopathological integrity of gastrostomy closure and the abdominal wall-ileum stoma tract formation were assessed 1 wk after the operation. Results Transgastric endoscopic tube ileostomy was successful in all six pigs, without major bleeding. The mean operating time was 71 min (range: 60-110 min). There were no intraoperative complications or hemodynamic instability. The post-mortem, which was conducted 1-wk postoperatively, showed complete healing of the gastrostomy and adequate stoma tract formation of ileostomy. Conclusion Transgastric endoscopic tube ileostomy is technically feasible and reproducible in an animal model, and this technique is worthy of further improvement.

The extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK-MAPK) is critical in human malignancies. It remained to be established whether DNA methyltransferases (Dnmt) and proliferating cell nuclear antigen (PCNA) involved in DNA methylation during RAF-transformed cell proliferation. The plasmid of constitutively active RAF was used to transfect gastric cell GES-1 and cancer cell AGS. RAF promoted cell proliferation, growth in soft agar and induced cell cycle progress faster than empty plasmid by accelerating G1/S transition in both cell lines, a massive induction of cyclin D1 and PCNA expression was observed, along with reduced expression of p16INK4A, p21WAF1 and p27KIP1. Methylation-specific polymerase chain reaction and bisulfite sequencing showed that the promoter of p16INK4A was methylated in RAF-transformed cells, treatment with 5-aza-dC or PD98059 restored the expression of p16INK4A, increased p21WAF1 and p27KIP1 partially, associated with upregulation of the activity of Dnmt in RAF-transformed cell GES-1, and also decreased the hypermethylation status of p16INK4A, but not all CpG islands of p21WAF1 and p27KIP1. These data suggest that RAF may induce cell proliferation through hypermethylation of tumor suppressor gene p16INK4A, while the epigenetic inactivation of p21WAF1 and p27KIP1 may be not a key factor in RAF-transformed cells.

Objective To investigate radiofrequency ablation of the slow pathway and significance of the different forms of slow potentials during radiofrequency catheter ablation of slow-pathway in patients with atrioventricular nodal reentrant tachycardia. Methods Analysed catheter radiofrequency ablation information of 54 patients with atrioventricular nodal reentrant tachycardia retrospectively and compared effective target rate and the success rate for the first time when using or not using slow potentials as ablation target and when using different forms of slow potentials as ablation target. Results Effective target rate was 71.4% and the success rate for the first time was 22.5% when using slow potentials as ablation target. Effective target rate was 85.7% and the success rate for the first time was 19.0% when using fragmentation-type slow potentials as ablation target. Effective target rate was 88.9% and the success rate for the first time was 38.9% when using first high frequency current and then low frequency current ablation of atrial double potential. Effective target rate was 10.0% and the success rate for the first time was 0% when firstly low frequency current and then high frequency current ablation of atrial double potential. Conclusion Effective target rate and the success rate for the first time was both high when using slow potentials as ablation target. Effective target rate and the success rate was both high when using fragmentation-type slow potentials and firstly high frequency current and then low frequency slow potentials as ablation target. The first low-frequency and then

The mechanical property of diamond-like carbon films with different hydrogen concentration is studied using molecular dynamical simulation based on REBO potential energy functions. The stiffness, hardness, and elastic modulus of the films are calculated from the load-unload curve. It is shown that both of the fraction of sp3 carbon and the concentration of H in the films have strong influence on the film hardness. When H concentration in the film is lower than 39%, the hardness is mainly determined by sp3 carbon, when H concentration further increases, the hardness decreases greatly despite the increase of the sp3 fraction.

Single-photon-counting imaging technology has been widely used in ultraweak bioluminescence, space environment detection and nuclear radiation detection. The structure and working principle of an ultraviolet single photon imaging system based on the microchannel plates and the wedge and strip anode are introduced. The gain characteristic of dark count of two and three microchannel plates is investigated under different voltages respectively. The experimental and fitted results show that the dark count pulse height distribution of microchannel plate has exponential distribution, and the dark count rate rises as the voltage increases. It is shown that the gain becomes more homogeneous as the voltage increases by measuring the pulse height distribution under different voltages. Then the effects of microchannel plate voltage and distance on system resolution with two microchannel plates are tested. It is revealed that the system resolution rises as the voltage of microchannel plate increases, and can be improved also by increasing the distance of microchannel plate properly.