Read-through circular RNA rt-circ-HS promotes hypoxia inducible factor 1α expression and renal carcinoma cell proliferation, migration and invasiveness / 北京大学学报(医学版)

OBJECTIVE@#To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.@*METHODS@#Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.@*RESULTS@#We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.@*CONCLUSION@#The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.

Study on transfusion-related acute lung injury caused by HLA-Ⅱ antibody / 中国输血杂志

【Objective】 To explore the risk factors of transfusion-related acute lung injury (TRALI). 【Methods】 The clinical symptoms, signs, imaging examinations, and laboratory test results of two patients with TRALI after blood transfusion were retrospectively analyzed, and human leukocyte antigen (HLA) genotyping of the patient and HLA antibodies typing of the plasma donors were performed. 【Results】 The clinical manifestations and laboratory parameters of two patients were consistent with those of TRALI after blood transfusion. After timely clinical respiratory support treatment, all patients were improved. Blood donors produced high titers of HLA-Ⅱ antibodies after pregnancy, including antibodies that specifically recognize the patient′s HLA antigen. 【Conclusion】 Two patients developed TRALI after platelet transfusion from a female blood donor, which was caused by HLA-Ⅱ antibodies.

Priming mechanism for the apoptosis of corneal endothelial cells induced by high pressure / 眼科新进展

Objective To investigate the initiation pathway of corneal endothelial cell apoptosis induced by high-pression.Methods Primary rabbit corneal endothelial cells were identified by immunohistochemistry and cultured under high pressure 50 mmHg (1 kPa =7.5 mmHg) for 1 h,2 h,24 h,respectively,while cells cultured under the normal pressure 15 mmHg served as the normal pression group.In addition,the first generation of rabbits corneal endothelial ceils with 70％ to 80％ fusion were pretreated with 10-6 mol · L-1 anti-Caspase 8 and anti-Caspase 9 for lh,followed by 50 mmHg pression for the treatment of the cells;while cells cultured with no inhibitor in the same pression served as the control group.Then the expression of P53 and Bcl-2 protein was detected by Western blot,and cytochrome C in rabbit corneal endothelial cells was determined by immunofluorescence staining in all groups.Results The expression levels of P53 in the 50 mmHg group were 0.651 +0.007,0.805 ±0.006 and 0.839 ±0.011 after 1 h,2 h,24 h high-pression respectively,which were significantly higher than those in the normal pressure group (0.033 ± 0.004),and the difference approached statistical significance (all P ＜ 0.01).The expression of P53 protein in corneal endothelial cells gradually increased as time went on,and the difference was statistically significant between each two time-points (all P ＜ 0.01).Moreover,the expression of Bcl-2 in the 50 mmHg pressure group was 0.590 ± 0.009,0.724 ± 0.005 and 0.34 ± 0.016,respectively,which was higher than that in the normal pressure group (0.081 ±0.013),with signifi cant difference (all P ＜ 0.01),and the difference approached statistical significance between each two time points in this group (all P ＜ 0.01).The expression level of P53 in anti-Caspase 9 and anti-Caspase 8 group was 0.535 ± 0.007 and 0.703 ± 0.010,respectively,which was significantly lower than that in the control group (0.727 ± 0.021),and the difference was statistically significant (all P ＜ 0.01).The expression of Bcl2 was 0.312 ± 0.003 and 0.442 ± 0.011,respectively,which were significantly lower than that in the control group (0.501 ± 0.011),with statistical difference (P ＜ 0.01).Finally,the expression of P53 and Bcl-2 in anti-Caspase 9 group was lower than that of anti-Caspase 8 group (P ＜ 0.01),indicating that anti-Caspase 9 had more enhanced inhibitory effect on the apoptosis of corneal endothelial cells than anti-Caspase 8.Conclusion AntiCaspase 9 inhibitor could effectively block the corneal endothelial cell apoptosis induced by high pressure.And the damage from high pressure on corneal endothelial cells mainly triggers the release of cytochrome C from chondriosome to activate the endogenous enzyme linked apoptotic pathway in which Caspase 9 involves.

Priming mechanism for the apoptosis of corneal endothelial cells induced by high pressure / 眼科新进展

Objective To investigate the initiation pathway of corneal endothelial cell apoptosis induced by high-pression.Methods Primary rabbit corneal endothelial cells were identified by immunohistochemistry and cultured under high pressure 50 mmHg (1 kPa =7.5 mmHg) for 1 h,2 h,24 h,respectively,while cells cultured under the normal pressure 15 mmHg served as the normal pression group.In addition,the first generation of rabbits corneal endothelial ceils with 70％ to 80％ fusion were pretreated with 10-6 mol · L-1 anti-Caspase 8 and anti-Caspase 9 for lh,followed by 50 mmHg pression for the treatment of the cells;while cells cultured with no inhibitor in the same pression served as the control group.Then the expression of P53 and Bcl-2 protein was detected by Western blot,and cytochrome C in rabbit corneal endothelial cells was determined by immunofluorescence staining in all groups.Results The expression levels of P53 in the 50 mmHg group were 0.651 +0.007,0.805 ±0.006 and 0.839 ±0.011 after 1 h,2 h,24 h high-pression respectively,which were significantly higher than those in the normal pressure group (0.033 ± 0.004),and the difference approached statistical significance (all P ＜ 0.01).The expression of P53 protein in corneal endothelial cells gradually increased as time went on,and the difference was statistically significant between each two time-points (all P ＜ 0.01).Moreover,the expression of Bcl-2 in the 50 mmHg pressure group was 0.590 ± 0.009,0.724 ± 0.005 and 0.34 ± 0.016,respectively,which was higher than that in the normal pressure group (0.081 ±0.013),with signifi cant difference (all P ＜ 0.01),and the difference approached statistical significance between each two time points in this group (all P ＜ 0.01).The expression level of P53 in anti-Caspase 9 and anti-Caspase 8 group was 0.535 ± 0.007 and 0.703 ± 0.010,respectively,which was significantly lower than that in the control group (0.727 ± 0.021),and the difference was statistically significant (all P ＜ 0.01).The expression of Bcl2 was 0.312 ± 0.003 and 0.442 ± 0.011,respectively,which were significantly lower than that in the control group (0.501 ± 0.011),with statistical difference (P ＜ 0.01).Finally,the expression of P53 and Bcl-2 in anti-Caspase 9 group was lower than that of anti-Caspase 8 group (P ＜ 0.01),indicating that anti-Caspase 9 had more enhanced inhibitory effect on the apoptosis of corneal endothelial cells than anti-Caspase 8.Conclusion AntiCaspase 9 inhibitor could effectively block the corneal endothelial cell apoptosis induced by high pressure.And the damage from high pressure on corneal endothelial cells mainly triggers the release of cytochrome C from chondriosome to activate the endogenous enzyme linked apoptotic pathway in which Caspase 9 involves.

Improved development of somatic cell cloned bovine embryos by a mammary gland epithelia cells in vitro model

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.

Evaluation of the effects of standard rescue procedure on severe trauma treatment in china / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study aimed to evaluate the effects of standard rescue procedure (SRP) in improving severe trauma treatments in China.</p><p><b>METHODS</b>This study was conducted in 12 hospitals located in geographically and industrially different cities in China. A standard procedure on severe trauma rescue was established as a general rule for staff training and patient treatment. A regional network (system) efficiently integrating prehospital rescue, emergency room treatments, and hospital specialist treatments was built under the rule for information sharing and improving severe trauma treatments. Treatment outcomes were compared between before and 1 year after the implementation of the SRP.</p><p><b>RESULTS</b>The outcomes of a total of 74,615 and 12,051 trauma cases were collected from 12 hospitals before and after the implementation of the SRP. Implementation of the SRP led to efficient cooperation and information sharing of different treatment services. The emergency response time, prehospital transit time, emergency rescue time, consultation call time, and mortality rate of patients were 24.24 ± 4.32 min, 45.69 ± 3.89 min, 6.38 ± 1.05 min, 17.53 ± 0.72 min, and 33.82% ± 3.87% (n = 441), respectively, before the implementation of the standardization and significantly reduced to 10.11 ± 3.21 min, 22.39 ± 4.32 min, 3.26 ± 0.89 min, 3.45 ± 0.45 min, and 20.49% ± 3.11%, separately (n = 495, P < 0.05) after that.</p><p><b>CONCLUSIONS</b>Staff training and SRP can significantly improve the efficiency of severe trauma treatments in China.</p>

Application of free flap pedicled with supracarpal cutaneous branch of ulnar artery in repairing of finger replantation / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate clinical application and clinical outcomes of free flap pedicled with supracarpal cutaneous branch of ulnar artery in repairing of finger replantation with skin defect.</p><p><b>METHODS</b>From April 2007 to March 2013,25 patients affected by finger amputation with skin defect were replanted and repaired by free flap pedicled with supracarpal cutaneous branch of ulnar artery. Among them, 18 patients were male and 7 were female,with an average age of 31.5 years old (ranged 16 to 58). The time of trauma to admission ranged from 45 to 210 min (averaged 105). Fifteen patients were complete separted, and 10 patients were non-complete separated. The area of flaps ranged from 3.5 cm x 2.0 cm to 4.5 cm x 3.0 cm, and the vessels were anastomosed through end-to-end. The functional evaluation standard of finger replantation was used to evaluate the postoperative function.</p><p><b>RESULTS</b>Twenty-four cases were finally survived. Two flaps occurred vascular crisisin within 48 h after operation, one of which was survived after anti-vasospasm treatment and changing dressing,another was replanted finger for failed to survive. One had infection and healed after changing dressing. Twenty-four cases were followed up from 3 to 38 months with an average of 16.5 months. The appearance and texture of flaps were satisfactory, and the superficial senses of pain and touch were recovered,and two-point discrimination was 5.5 to 11 mm (averaged 7.4 mm). According to functional evaluation standard finger replantationissued by Hand Surgery Association of Chinese Medical Association, 8 cases got excellent results, 14 good and 2 poor.</p><p><b>CONCLUSION</b>The free flap pedicled with supracarpal cutaneous branch of ulnar artery can be used in complex finger replantation with skin and vessels defect, which can extend operation indications, recover function and appearance for maximum.</p>

Clinical application of reverse radial hypothenar flap for finger soft tissue defect / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effect of reverse radial hypothenar flap for finger soft tissue defect.</p><p><b>METHODS</b>From Mar. 2006 to Mar. 2010, 13 cases (14 fingers) with finger soft tissue defects were treated with reverse radial hypothenar flaps pedicled with ulnar palmar digital artery of little finger. The defects were 1.9 cm x 1.5 cm -4.0 cm x 2.0 cm in size. The flap size ranged from 1.5 cm x 2.0 cm to 4.0 cm x 2.0 cm.</p><p><b>RESULTS</b>All the flaps survived completely with primary healing both in donor and recipient area. 12 cases (13 fingers) were followed up for 1-3 years. The flaps color was similar to the unaffected fingers. Cicatricial contracture happened in one case due to contracture of palmar fascia. The two-point discrimination distance on flap was 3.2-5.3mm. The active and passive movement of finger joints was evaluated as excellent in 12 fingers, good in one finger. There was no complaint about the feeling at the donor site. Two months after operation, all patients could go back to work.</p><p><b>CONCLUSIONS</b>The reverse radial hypothenar flap is very suitable for finger soft tissue defect with less morbidity to donor site.</p>

Somatic nuclear transplantation and serial nuclear transplantation of human finger-domain lacking t-PA gene in goat / 生物工程学报

In order to research developmental competence of transgenic somatic cell by serial nuclear transplantation, goat cloned embryos were compared with recloned embryos in ability of in vitro development. Fetal fibroblasts including human finger-domain lacking t-PA gene was microinjected into cytoplasm of the MII oocytes. Goat embryos (G0) were cloned by this procedure. A single blastomere from 16 - 64-cell goat cloned embryos (G0) was microinjected into Intracytoplasm of the MII oocytes. Goat embryos (G) were cloned by this procedure. Goat embryos (G2, G3) were recloned by using 16 - 64-cell recloned embryos. The developmental time of donor embryo affected the developmental rate of recloned embryos (G1, G2). The results show: the cleavage rate of cloned embryos (G0) (76.45% +/- 1.17%) was no difference significantly with recloned embryos (G1 G2 G3) (72.18% +/- 1.97%, 76.05% +/- 2.38%, 75.99% +/- 2.84%); the developmental rate of morulae and blastocysts of cloned embryos (47.20% +/- 2.93%, 11.00% +/- 1.42%) were higher than these of recloned embryos(34.99% +/- 2.66%, 28.23% +/- 2.00%, 23.34% +/- 1.99%) (3.87% +/- 0.67%, 2.08% +/- 1.66%, 0); the morulae rate(29.57% +/- 1.53%, 24.43% +/- 1.87%) and blastocysts rate(1.96% +/- 1.31%, 2.01% +/- 1.34%) of recloned embryos (G1 G2) from 16-cell recloned embryos were lower than those(34.32% +/- 1.31%, 29.76% +/- 1.66% and 3.86% +/- 1.03%, 3.48% +/- 0.34% )from 32 - 64-cell recloned embryos (P > 0.05). In conclusion, nuclear transfer embryos should not were recloned mostly; and the embryos recloned by using 32 - 64-cell embryos achieved higher developmental ability compared with using 16-cell embryos.

Role of the central interleukin-1 in stress responses / 生理学报

The molecules of interleukin-1 (IL-1) system are widely distributed in central nervous system. As a classical pro-inflammatory factor, central IL-1 has diverse biological functions and plays a pivotal role in a number of important physiological and pathophysiological processes. During the past few years, particular attentions have been directed to the stress mediator actions of central IL-1. This paper reviews some recent findings in the studies of central IL-1 functions in stress responses, including the effects of stress on central IL-1, the roles of IL-1 in the initiation of stress responses, the neural circuitries and intracellular signal transduction pathways involved in the central IL-1 mediated stress responses, as well as the actions of central IL-1 on brain high function and behavior under stressful conditions.