Objective To investigate the mid- to long-term follow-up results of ascending aorta (AAO)-descending thoracic aorta (DTA) bypass grafting via median sternotomy incision for the treatment of complex aortic arch coarctation. Methods A retrospective analysis was conducted on the clinical data of patients with complex aortic arch coarctation who underwent AAO-DTA bypass grafting via median sternotomy incision at the First Hospital of Tsinghua University from August 2004 to May 2017. Results A total of 7 patients were enrolled, including 4 males and 3 females, aged (13.3±4.6) years, and weighted (40.2±12.2) kg. Six (85.7%) patients had concomitant upper limb hypertension. Four patients were aortic arch coarctation combined with intracardiac malformations, two were post-operative restenosis, and 1 was post-operative restenosis combined with intracardiac malformation. All patients underwent surgery under cardiopulmonary bypass. There were no perioperative deaths or major complications. The pre-operative upper-lower limb pressure difference was (39.3±19.2) mm Hg, which decreased to (2.9±2.7) mm Hg post-operatively (P<0.01). The follow-up period was (14.9±5.9) years. There were no long-term deaths or artificial graft-related complications. Except for one patient who still had mild hypertension, the blood pressure of the remaining patients returned to normal. Conclusion AAO-DTA bypass grafting via median sternotomy incision for the treatment of complex aortic arch coarctation can effectively reduce upper limb blood pressure and the upper-lower limb arterial pressure difference, has fewer complications, and demonstrates satisfactory mid- to long-term efficacy.

Animals ; Bone Morphogenetic Protein 4 ; Cardiovascular Diseases ; Endothelial Cells ; Fibroblasts ; Humans ; In Vitro Techniques ; Induced Pluripotent Stem Cells ; Lipoproteins ; Mesoderm ; Methods ; Mice ; Models, Animal ; Pluripotent Stem Cells ; Swine

Regenerative therapy holds great promise in the development of cures of some untreatable diseases such as cardiovascular diseases, and pluripotent stem cells (PSCs) including induced PSCs (iPSCs) are the most important regenerative seed cells. Recently, differentiation of human PSCs into functional tissues and cells in vitro has been widely reported. However, although porcine reports are rare they are quite essential, as the pig is an important animal model for the in vitro generation of human organs. In this study, we reprogramed porcine embryonic fibroblasts into porcine iPSCs (piPSCs), and differentiated them into cluster of differentiation 31 (CD31)-positive endothelial cells (ECs) (piPSC-derived ECs, piPS-ECs) using an optimized single-layer culture method. During differentiation, we observed that a combination of GSK3β inhibitor (CHIR99021) and bone morphogenetic protein 4 (BMP4) promoted mesodermal differentiation, resulting in higher proportions of CD31-positive cells than those from separate CHIR99021 or BMP4 treatment. Importantly, the piPS-ECs showed comparable morphological and functional properties to immortalized porcine aortic ECs, which are capable of taking up low-density lipoprotein and forming network structures on Matrigel. Our study, which is the first trial on a species other than human and mouse, has provided an optimized single-layer culture method for obtaining ECs from porcine PSCs. Our approach can be beneficial when evaluating autologous EC transplantation in pig models.

Regenerative therapy holds great promise in the development of cures of some untreatable diseases such as cardiovascular diseases, and pluripotent stem cells (PSCs) including induced PSCs (iPSCs) are the most important regenerative seed cells. Recently, differentiation of human PSCs into functional tissues and cells in vitro has been widely reported. However, although porcine reports are rare they are quite essential, as the pig is an important animal model for the in vitro generation of human organs. In this study, we reprogramed porcine embryonic fibroblasts into porcine iPSCs (piPSCs), and differentiated them into cluster of differentiation 31 (CD31)-positive endothelial cells (ECs) (piPSC-derived ECs, piPS-ECs) using an optimized single-layer culture method. During differentiation, we observed that a combination of GSK3β inhibitor (CHIR99021) and bone morphogenetic protein 4 (BMP4) promoted mesodermal differentiation, resulting in higher proportions of CD31-positive cells than those from separate CHIR99021 or BMP4 treatment. Importantly, the piPS-ECs showed comparable morphological and functional properties to immortalized porcine aortic ECs, which are capable of taking up low-density lipoprotein and forming network structures on Matrigel. Our study, which is the first trial on a species other than human and mouse, has provided an optimized single-layer culture method for obtaining ECs from porcine PSCs. Our approach can be beneficial when evaluating autologous EC transplantation in pig models.

Objective: To explore drug utilizing regularity of traditional Chinese medicine(TCM) in treating and preventing asthenopia by analyzing the patent status of TCM in the field of asthenopia control for nearly 20 years. Method: Global patents about TCM in treating and preventing asthenopia were systematic searched in IncoPat platform.The application trend,legal status and categories of patents were analyzed.Meanwhile,the oral prescriptions and external prescriptions were performed correlation index analysis by IBM SPSS Modeler 18.0,respectively;the difference of compatibility rules between them was compared. Result: The number of global patents in treating and preventing asthenopia gradually increased,and the proportion of patents from China was more than 99%.The main patent applications were pharmaceutical composition,oral preparation,external eye patch and so on,but the ratio of licensed patents in total patents was low.In term of drug utilizing regularity,the oral prescriptions paid much attention to using TCM for nourishing the liver and kidney,while external prescriptions highlighted relieving sickness heat and detoxification. Conclusion: Patents of TCM in treating and preventing asthenopia has been kept increasing in recent years,but the authorization rate is low.Formulation based on TCM theory can be statistically summarized,which can be helpful for the development of anti-asthenopia products.

The clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile genome editing tool with high efficiency. A guide sequence of 20 nucleotides (nt) is commonly used in application of CRISPR/Cas9; however, the relationship between the length of the guide sequence and the efficiency of CRISPR/Cas9 in porcine cells is still not clear. To illustrate this issue, guide RNAs of different lengths targeting the EGFP gene were designed. Specifically, guide RNAs of 17 nt or longer were sufficient to direct the Cas9 protein to cleave target DNA sequences, while 15 nt or shorter guide RNAs had loss-of-function. Full-length guide RNAs complemented with mismatches also showed loss-of-function. When the shortened guide RNA and target DNA heteroduplex (gRNA:DNA heteroduplex) was blocked by mismatch, the CRISPR/Cas9 would be interfered with. These results suggested the length of the gRNA:DNA heteroduplex was a key factor for maintaining high efficiency of the CRISPR/Cas9 system rather than weak bonding between shortened guide RNA and Cas9 in porcine cells.
Base Sequence ; Complement System Proteins ; CRISPR-Cas Systems ; DNA ; Genome ; Nucleotides ; RNA, Guide ; Swine

Objective To explore the prognostic values of telomerase reverse transcriptase promoter (TERTp) mutation and 1p/19q co-deletion in newly-diagnosed O6-methylguanine-DNA methyltransferase (MGMT) promoter un-methylated/isocitrate dehydrogenase (IDH) wild-type glioblastoma multiform (GBM). Methods A total of 82 patients pathologically newly-diagnosed MGMT promoter un-methylated/IDH wild-type GBM, admitted to our hospitals from March 2016 to November 2018, were included in this study. TERTp mutations (TERTp wild-type and TERTp mutation [C228 mutation and C250 mutation]) in GBM specimens were detected by PCR sequencing, 1p/19q co-deletion in GBM specimens was detected by fluorescence in situ hybridization (FISH), and clinical data, adverse reactions and prognoses of patients with different molecular typing were compared. Results There were 33 patients in the TERTp wild type group with mean age of 48 years, and 49 patients in the TERTp mutation group with mean age of 59 years; the difference of age was significant (P<0.05); there were no statistical differences in gender distribution, Karnofsky performance status (KPS) scores, tumor sites and surgical resection degrees between the two groups (P>0.05). There were 8 patients with 1p/19q co-deletion and 74 patients without 1p/19q co-deletion; no significant differences in above clinical parameters were noted between the two groups. There were no statistically significant differences in the incidences of bone marrow suppression, digestive tract response and fatigue, disease progression rate, or survival rate between patients from TERTp wild type group and TERTp mutation group, and between patients with 1p/19q co-deletion and patients without 1p/19q co-deletion (P>0.05). No significant differences in above clinical parameters, disease progression rate, and survival rate were noted between patients with C228 mutation and C250 mutation (P>0.05). Conclusion TERTp typing and 1p/19q co-deletion status do not have prognostic value in newly-diagnosed MGMT un-methylated/IDH wild-type GBM patients; patients with TERTp mutations have older age than wild-type patients; patients with C250 mutation trend to have higher survival rate than those with C228 mutation.

Objective To investigate the prognostic influence factors of Solitaire stent thrombectomy in patients with acute anterior circulation macrovascular occlusion. Methods From March 2015 to March 2017,222 consecutive patients with acute anterior circulation macrovascular occlusion admitted to the Department of Neurosurgery,the 101stHospital of People′s Liberation Army and the Nanjing Jinling Hospital were enrolled retrospectively.They were all confirmed by DSA and were treated with Solitaire stent thrombectomy. According to the modified Rankin Scale(mRS) scores at 90 d after treatment,they were divided into a good prognosis group (0-2,n=120) and a poor prognosis group (3-6,n =102). The baseline data and clinical data of the two groups of patients were analyzed,including the risk factors for cardiocerebrovascular diseases,baseline National Institutes of Health Stroke Scale (NIHSS) score,occlusion sites (internal carotid artery or middle cerebral artery occlusion),collateral compensatory,onset to puncture time, operation time,onset to recanalization median time,recanalization status,preoperative Alberta stroke programme early CT score(ASPECTS),and symptomatic cerebral hemorrhage,and then further multivariate logistic regression analysis was conducted for the prognostic factors of patients. Results (1) The rate of good prognosis was 54.1% (120/222).There were no significant differences in patients′ age,NIHSS at admission,ASPECTS at admission,sex,hypertension,occlusion site,and rate of good collateral branches in both groups(all P<0.05).There were no significant differences in other baseline data (all P >0. 05). (2) Onset to puncture time and onset to successful recanalization median time of the patients in good prognosis group was lower than that of the poor prognosis group (182 [138,230]min vs.236[170,305]min, 237[175,269]min vs.288[223,367]min).The proportion of successful recanalization was higher than that of the poor prognosis group (98.3% [118/120] vs.78.4% [80/102]).The proportion of postoperative symptomatic intracerebral hemorrhage was lower than that of the poor prognosis group (2.5% [3/120] vs.21.6% [22/102]).There was significant difference between the two groups (all P <0.01). There was no significant difference in operative time between the two groups (P >0.05). (3)In the single factor analysis,the parameter of P <0.05 was used as an independent variable,and prognosis was used as a dependent variable,multivariate logistic regression analysis showed that the increased age (OR,1.096,95% CI 1.050-1.144),history of hypertension (OR,8.401,95% CI 2.960-23.845),increased baseline NIHSS score (OR,1.071,95% CI 1.007-1.138),prolonged onset to successful recanalization time (OR,1.019,95% CI 1.003-1.035),symptomatic intracerebral hemorrhage after procedure (OR,18.110,95% CI 4.656-70.434) were all the risk factors for poor prognosis(all P<0.05);higher ASPECTS score at admission(OR,0.641,95% CI 0.451-0.911) and successful recanalization (OR,0.127,95% CI 0.024-0.664) were all the protective factors of good prognosis (all P<0.05). Conclusions Higher ASPECTS at admission and successful recanalization were the protective factors of poor prognosis of Solitaire stent thrombectomy in patients with acute anterior circulation macrovascular occlusion.Increased age,history of hypertension,increased baseline NIHSS score,prolonged onset to successful recanalization time,and symptomatic intracerebral hemorrhage after procedure were the risk factors for poor prognosis of Solitaire stent thrombectomy in patients with acute anterior circulation macrovascular occlusion.

Objective To investigate the effect of bone cement distribution on efficacy of vertebroplasty.Methods From January 2013 to June 2016,a total of 132 cases (132 vertebrae) with single segment osteoporotic thoracolumbar vertebral fractures underwent parallel vertebroplasty surgery,and there were 57 male,75 female,with an average age of (71.6±2.2) years old (ranged from 65 to 86 years old).On the basis of the postoperative X-ray films of bone cement distribution were divided into 3 groups.The bone cement was biased to the lateral side of the vertebral body (partial group,35 cases),the bone cement was over the vertebral midline,but not completely filled with contralateral vertebral body (near midline group,46 cases),and the bone cement was filled with bilateral vertebral body (bilateral group,51 cases).There were 15 males and 20 females in the partial group,aged (70.3±5.3) years old;20 males and 26 females in the proximal midline group,aged (72.1±3.2) years old;22 males and 29 females in the bilateral group,aged (71.2±4.6) years old.Local anesthesia was used to make the patient prone to operate on the operating bed.The head and tail of the bed were increased at the same time slightly and vertebral compression fractures reduction was performed.Bone cement was injected into the vertebral body through partial or bilateral transpedicular approach.The visual analogue scores (VAS) were measured of preoperation,postoperation and 3,6,12 months after surgery.Analysis of variance for each group and VAS before and after operation,and postoperative complications were observed too.Results All the 132 cases were followed up for 1-12 months,with an average of (11±0.3) months.There were statistically significant differences in the immediate effect of postoperation among partial group,near middle group and the bilateral group (F=90.472,P=0.000),VAS score in partial group was lower than that in bilateral group (t=11.433,P=0.000),but higher than that in near midline group (t=11.106,P=0.000),and the differences were statistically significant,but there was no significant difference between near midline group and bilateral group (t=0.581,P=0.563).VAS score showed no statistically difference among the three groups 3 months,6 months and 1 year follow-up after operation (F=0.892,P=0.413;F=0.342,P=0.713;F=0.834,P=0.441).In 3 eases of partial group,the pain was not relieved due to unfilled cement until the contralateral bone was injected into the bone cement.However,there were 11 cases of cement leakage in partial group,13 cases in near midline group,and 3 cases in bilateral group.Conclusion The distribution of bone cement is one of the main factors affecting the clinical efficacy after vertebroplasty,and the clinical effect of distributing the midline of vertebral body is better than the one side.

Vigorously development of the health service industry has received great attention,but the health service industry faces many difficulties.By analyzing the current predicament,this paper put forward the KY3H health care service model and systematically analyzed its six innovations:from "disease as the center" to "health as the center",from "homogenized group health service" to "Personalized and Humanized Health Service",from "Extensive Health Service"to "Precise Intervention of Individual Health Status",from "Single Health Service" to "Trinity Service of Health,Culture,Health Management and Health Insurance",from the "seeking medical" service model to "seeking one's own"service model,as well as from the "fixed (fixed-time,fixed-point,fixed-mode) service" to "anytime,anywhere full-time service".This model has achieved initial success in practice.It is an important way to solve the dilemma.