Objective To evaluate the clinical features and curative effect of patients with Takayasu's arteritis.Methods The clinical data of fifty-eight patients,which were divided into different groups according to gender,clinical classifications,disease activity,and so on,were retrospectively analyzed,to compare whether the results of clinical features,laboratory tests or prognosis had significant differences.Results The data suggested the fact that stroke as first manifestation(2/9 vs 0/49,P =0.022) and incidence of hypertension (9/9 vs 28/49,P =0.037),as well as critical hypertension(8/9 vs 18/49,P =0.011) were more common in male patients than in female patients.The active Takayasu's arteritis patients showed that the level of erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),platelet and fibrinogen was elevated,while the level of Albumin/Globulin ratio was decreased.Electrophoresis showed that the elevation of globulin mainly based on globulin α1 and globulin α2 (P ＜ 0.05).The reduction of complement C3 is more common in inactive group than in active group.Conclusion Stroke and hypertension are more common in male Takayasu's arteritis patients than in female patients.The active Takayasu' s arteritis patients showed elevated level of ESR,CRP,platelet and fibrinogen,while the level of Albumin/Globulin ratio was reduced.

Background:The reproductive health addresses the reproductive processes,functions and system at all stages of life.Enhancing the level of global reproductive health is the goal of sustained attention and struggle by the international community.The social and economic development in Southeast Asia is lagging behind,and its female reproductive health is worrying,while the differences of female reproductive health among different regions are significant.Objective:To obtains the necessity and urgency of strengthening the reproductive health level of Southeast Asian countries,so as to provide the basis for the priorities and target to policy-makers and health administrators to improve reproductive health.Methods:Literature review were searched in PubMed,Web of Science databases,Google scholar database,and WHO's webpages.Maternal mortality ratio,contraceptive rates,unmet need for family planning,antenatal and postnatal care coverage,and sexually transmitted disease were the five key indicators and the influence factors for female reproductive health status in Southeast Asian countries.Results:The reproductive health of Southeast Asian women were still at a lower level overall and varied in different regions and conntries.Women's education and attitude,accessibility of service,socioeconomic and cultural factors,etc.were the potential influencing factors.Conclusion:There is left quite large space for improvement to the reproductive health in Southeast Asian countries and efficient interventions can be achieved for the key and easier-improved risk factors such as education and in high-risk areas.

Objective To introduce and discuss the efficacy of a new technique to perform transperitoneal single-docking robot-assisted laparoscopic nephroureterectomy (RNU).Methods A total of 44 patients diagnosed with urothelial neoplasm of the renal pelvis or were investigated from January 2016 to November 2019.RNU was performed by a single surgeon.Among the 44 patients,31 were male,and 13 were female.The median age was 63 (IQR:58-71).The median body mass index (BMI) was 23.08 (IQR:21.55-24.60) kg/m2.All operations were performed with general anesthesia.The patients were positioned 80 degrees flank with the diseased side up,and the head was tilted 10 degrees downwards.The camera port was placed one finger lateral to the umbilicus.For the right-sided tumors,robotic arm 1 was inserted through the trocar on the right pararectus line,8 cm above the umbilicus,and robotic arm 2 was inserted through the trocar on the same line,8 cm below the umbilicus.Assistant trocar 1 was placed where the anterior midline joins the perpendicular bisector of the camera port and robotic 2,and assistant trocar 2 was placed below the xiphoid process.For the left-sided tumors,all trocars were centrosymmetric to that of the right-sided tumors,except that assistant port 2 was placed 3 finger width above the pubic symphysis.The peritoneum was incised along the Toldt line,and the inferior vena cava was isolated (for left sided tumor,the abdominal aorta was isolated instead).The renal artery and vein were clipped with Hem-o-lok and ligated,and the kidney were isolated.The ureter was identified and isolated downwards across the common iliac artery and then clipped distal to the tumor site.The bladder cuff was resected and sutured under the laparoscopy.Results The median operation time was 145 (IQR:130-175) min,with the median console time of 119 (IQR:108.5-136.0) min,the anastomosis of bladder cuff of 12 min,and the median estimated blood loss of 50 (20-100)ml.After the surgery,6 Clavien-Dindo grade 2 complications occurred,including 2 chylous leakage,1 hemostasis,1 blood transfusion,1 deep vein thrombus,and 1 acute coronary syndrome.The median length of stay (LOS) was 8 (IQR:6.5-10.0) d.The median length of follow-up was 12 months.In total,5 patients were dead,including 3 cancer-specific death.Four recurrence occurred and caused 3 death.The 2-year overall survival and progression-free survival were 68.2％ and 77.9％,respectively.Conclusions The technique of RNU with simultaneous bladder cuff excision (BCE).Our technique improved the surgical outcome.The perioperative complication rate was low,and the short-term survival outcomes were satisfactory.

Objective To investigate the effect of intravenous tranexamic acid (TXA) on perioperative hidden blood loss in percutaneous pedicle screw fixation for thoracolumbar fractures.Methods A prospective study was conducted in the 113 patients who would be subjected to percutaneous pedicle screw fixation for thoracolumbar fracture from January 2017 to December 2017.They were randomly assigned into an observation group (n =58) receiving intravenous drip of 15 mg/kg TXA 30 minutes preoperation or a control group (n =55) receiving intravenous drip of normal saline solution 30 minutes preoperation.The total blood loss and hidden blood loss 24 hours postoperation,D-dimer volume,incidences of deep vein thrombosis and other complications were recorded and compared between the 2 groups.Results There were 54 patients in the observation group and 50 patients in the control group for statistic analysis.The observation group had significantly less total blood loss (319.0 ± 140.5 mL) and hidden blood loss (242.0 ± 143.4 mL) 24 hours postoperation than the control group (418.7 ± 188.1 mL and 354.7 ± 181.9 mL,respectively) (P ＜ 0.05).There were no significant differences between the 2 groups in operation time or intraoperative blood loss (P ＞ 0.05).The volume of postoperative D-dimer was significantly higher than the preoperative value in both groups (P ＜ 0.05).No thromboembolic events occurred in either group.Conclusion Intravenous TXA may significantly reduce intraoperative hidden blood loss with no increased rik of thromboembolic events in percutaneous pedicle screw fixation for thoracolumbar fractures.

Objective To compare minimally invasive percutaneous pedicle screw fixation and open pedicle screw fixation for neurologically intact thoracolumbar fractures.Methods A retrospective study was conducted in the 180 patients who had been treated for thoracolumbar fractures without neurological deficits from January 2016 to December 2016.Of them,93 were treated by minimally invasive percutaneous pedicle screw fixation and 87 by open pedicle screw fixation.The 2 groups were compared in terms of blood loss,radiological parameters,visual analogue scale (VAS) and Oswestry disability index (ODI).Results Compared with the open surgery group,the minimally invasive surgery group had significantly shorter operating time (95.8 ±33.4 min versus 106.3 ±30.9 min),significantly less intraoperative blood loss (65.8 ±40.3 mL versus 183.1 ± 77.5 mL),significantly less total blood loss in theory 24 hours after surgery (374.7 ± 160.6 mL versus 614.8 ± 242.6 mL) and significantly shorter hospital stay (5.2 ± 2.0 d versus 6.7 ± 2.7 d),but significantly longer C-arm exposure time (23.6 ±4.2 min versus 12.4 ±4.1 min) and significantly more hidden blood loss 24 hours after surgery (308.9 ± 159.0 mL versus 243.5 ± 195.5 mL) (P ＜ 0.05).Compared with preoperation,significant improvements were observed at one week postoperation and the last follow-up in the 2 groups regarding the percentage of anterior height of the fractured vertebral body and cobb angle (P ＜ 0.05),but there were no significant differences in the percentage of anterior height of the fractured vertebral body or cobb angle between the 2 groups at one week postoperation or at the last follow-up (P ＞ 0.05).At 3 days postoperation,significant better pain relief was observed in the minimally invasive surgery group than in the open surgery group (P ＜ 0.01),but at the last follow-up no obvious pain was reported in either group.At the last follow-up,there was no significant difference between the 2 groups in ODI (6.2 ± 1.1 versus 6.0 ± 1.4) (P =0.320).Conclusions In the treatment of neurologically intact thoracolumbar fractures,minimally invasive percutaneous pedicle screw fixation may lead to shorter operating time,less blood loss and shorter hospital stay but no poorer radiological outcomes or long-term patient-reported outcomes than the open pedicle screw fixation.However,it should be noted that the former may lead to a higher volume of hidden blood loss.

Objective:To construct a double-layer bone-on-a-chip containing bone matrix, with which the process of osteoblast and osteoclast differentiation in vitro is stimulated, aiming to provide a new platform for the development of osteoporosis medications. Methods:Software WorkSoild was used to design the double-layer and double-channel bone-on-a-chip and the template was fabricated by photolithography. With polydimethylsiloxane (PDMS) as the raw material, the main body of the chip was prepared by mold fabrication. The inlets and outlets of the four channels of the culture room were separated with bovine cortex bones and sealed with liquid storage columns. In the chip verification experiment, chips were divided into osteogenic and osteoclastic induction groups and osteogenic and osteoclastic control groups. In the osteogenic and osteoclastic induction groups, precursor cells of mouse embryonic osteoblast, MC3T3-E1 and mouse macrophage RAW264.7 were inoculated on the chip separately. Osteogenic induction lasted 14 days and osteoclastic induction 7 days. MC3T3-E1 cells and RAW264.7 cells were not induced in the osteogenic and osteoclastic control groups. The following indicators were observed: (1) Appearance and sealing performance of the chip: After the chip was prepared, photos were taken to observe its appearance and sealing tests were conducted to observe its sealing performance. (2) Biocompatibility: At 3 days after MC3T3-E1 cells were inoculated onto the chip and cultured and at 1, 3 and 5 days after RAW264.7 cells were inoculated onto the chip and cultured, the cell survival was observed with calcein acetoxymethyl ester/propidium iodide (AM/PI) staining and Cell Counting Kit 8 (CCK-8). (3) Osteogenic differentiation: Alkaline phosphatase (ALP) staining and alizarin red staining were performed on the cells in the osteogenic induction group to observe the osteogenic induction. RNA was collected from the osteogenic induction group and the osteogenic control group, the expression of osteoblast marker Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) and type I collagen (COL1A1) was detected by real-time florescent quantitative PCR (qPCR), and the differentiation degree and osteogenic ability of osteoblasts were observed. (4) Osteoclast differentiation: tartrate-resistant acid phosphatase (TRAP) staining was performed on cells in the osteoclastic induction group to observe osteoclast differentiation. RNA was extracted from the osteoclastic induction group and the osteoclastic control group for qPCR of osteoclast differentiation-related genes, and the expression levels of the osteoclast marker gene TRAP, cathepsin K (CTSK) and dendritic cell specific transmembrane protein (DC-STAMP) were detected.Results:The double-layer bone-on-a-chip containing bone matrix was 3 cm×3 cm in size and transparent as a whole. The structure of the system on the chip system was compact and had no seepage. It was shown by calcein AM/PI staining that at 3 days after MC3T3-E1 cells and RAW264.7 cells were cultured, very few red fluorescent dead cells were found. CCK-8 test showed that within 5 days after being cultured, the cell viability was all above 90%, indicating that the biocompatibility of the chip was good and the cells could survive and proliferate normally. The results of ALP and alizarin red staining showed that MC3T3-E1 cells successfully differentiated into osteoblasts and produced calcified nodules in the osteogenic induction group at 14 days after the induction. The qPCR results showed that the relative expression level of RUNX2 in MC3T3-E1 cells in the osteogenic induction group was 4.98±0.74, which was significantly higher than that of the control group (0.99±0.03) ( P<0.01). The relative expression level of OCN in MC3T3-E1 cells was 7.98±0.76, which was significantly higher than that of the control group (1.00±0.06) ( P<0.01). The relative expression level of COL1A1 in MC3T3-E1 cells was 7.07±0.56, which was significantly higher than that of the control group (0.97±0.03) ( P<0.01). The TRAP staining results showed that the RAW264.7 cells in the osteoclastic induction group differentiated to giant multinucleated osteoclasts, and TRAP protein was expressed in large quantity in the osteoclasts. The results of qPCR showed that the relative expression level of TRAP in RAW264.7 cells in the osteoclastic induction group was 3.35±0.37, which was significantly higher than that of the control group (1.01±0.06) ( P<0.01). The relative expression level of CTSK in RAW264.7 cells was 3.46±0.79, which was significantly higher than that of the control group (1.01±0.05) ( P<0.01). The relative expression level of DC-STAMP in RAW264.7 cells was 1.92±0.12, which was significantly higher than that of the control group (0.98±0.08) ( P<0.01). Conclusions:The double-layer bone-on-a-chip containing bone matrix is compact in structure, can be cultured in vitro for a long time, has good biocompatibility and can be used for inducing osteogenic and osteoclast differentiation. Therefore, it is expected to provide a new research platform for exploring the mechanism of osteoporosis and medication screening.

Objective:To design and construct a bone nonunion organoid on chip and explore the mechanism of aseptic bone nonunion.Methods:First a semi-open microfluidic chip was designed, on which human bone marrow mesenchymal stromal cells (BMSC), human fetal lung fibroblast 1, (HFL1) and human umbilical vein endothelial cells (HUVEC) were co-cultured, and a three-dimensional organ on chip system was established. Different proportions of HFL1 and HUVEC were co-cultured with BMSC, which were divided into the control group (HFL1∶HUVEC=1∶1), the fibrosis group (HFL1∶HUVEC=3∶1) and the vascularization group (HFL1∶HUVEC=1∶3). The osteogenic differentiation of BMSC was observed by alkaline phosphatase (ALP) and Alizarin red staining. The transcription level of osteogenic marker genes SP7, RUNX2, ALPL, and BGLAP, and vascularization related genes KDR and VWF were analyzed by qPCR. The expression levels of RUNX2 and ALP were determined by Western Blot. Results:In the co-culture system of BMSCs, HFL1, and HUVECs, BMSCs exhibited normal growth and apparent biomineralization behavior. Endothelial cells were capable of forming structured vascular networks, confirming the successful establishment of the system. Compared to the baseline group, the fibrotic group showed no significant decrease in BMSC osteogenic differentiation. The relative expression levels of the mineralization marker genes ALPL and BGLAP were 0.55±0.19 ( P<0.001) and 0.42±0.27 ( P<0.001), respectively. Vascularization genes KDR and VWF were downregulated, with relative expression levels of 0.49±0.17 ( P<0.001) and 0.49±0.21 ( P<0.001). In contrast, in the vascularized group, BMSC osteogenic differentiation genes SP7, RUNX2, ALPL, and BGLAP were upregulated, with relative expression levels of 2.91±0.52 ( P<0.001), 3.83±1.87 ( P<0.001), 3.22±1.29 ( P<0.001), and 5.21±1.46 ( P<0.001), respectively. Vascularization genes KDR and VWF were also upregulated, with relative expressions of 8.24±2.84 ( P<0.001) and 5.32±1.67 ( P<0.001). Western blot results indicated increased expression of RUNX2 and ALP in the vascularized group and decreased expression in the fibrotic group. Conclusion:The bone nonunion organoid on chip could partially simulate the local microenvironment of bone nonunion. Fibrosis may lead to a significant decrease in bone formation ability and vascularization level, which might be an important reason for the occurrence of aseptic bone nonunion.

ObjectiveThe biosynthetic pathways of benzylisoquinoline alkaloids（BIAs） in Nelumbo nucifera are of great theoretical and economic value. In this paper， N. nucifera O-methyltransferase（NnOMT） and N. nucifera N-methyltransferase（NnNMT） gene families were identified and analyzed by bioinformatics in order to facilitate the biosynthetic pathway of BIAs in N. nucifera. MethodBased on the whole genome of N. nucifera， UniPort and National Center for Biotechnology Information（NCBI） databases were used to identify the NnOMT and NnNMT gene families of N. nucifera， and analyze their physicochemical properties and subcellular localization， then TBtools， MEME， MEGA 11.0， FigTree 1.4.4 and other tools were used to analyze the phylogeny， sequence characteristics， gene structure， functional annotation and cis-acting elements of NnOMT and NnNMT genes identified in the previous stage. ResultA total of 61 NnOMT and NnNMT genes were identified in this paper， the number of amino acids encoded by these genes ranged from 168 aa to 580 aa， the isoelectric point ranged from 4.76 to 9.16， and the relative molecular weight ranged from 18 699.52 Da to 64 934.53 Da， most of which showed acidic and mostly hydrophilic proteins. There were 10 conserved motifs， Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis enriched a total of 12 pathways， including metabolism， biosynthesis of phenylpropane and isoquinoline alkaloids， etc. And Visualization of Gene Ontology（GO） enrichment results showed that 61 NnOMT and NnNMT genes were annotated to 32 items， which included 16 molecular functions［such as reduced nicotinamide adenine dinucleotide（NADH） activity and exopeptidase activity］ and 16 biological processes（such as metabolic process of carbon tetrachloride， anaerobic carbon tetrachloride metabolic process and responses to exogenous biological stimuli）. There were a variety of cis-acting elements in the promoter regions of NnOMT and NnNMT genes， mainly promoter and enhancer regions element， light responsive element and methyl jasmonate responsive element. ConclusionIn this study， a comprehensive bioinformatics analysis of 61 NnOMT and NnNMT genes is carried out based on the genome data of N. nucifera， which lays a foundation for research on the gene structure and function of NnOMT and NnNMT gene families， and provides a reference for biosynthetic pathway elucidation of BIAs in N. nucifera.

Many human genetic diseases, including Hutchinson-Gilford progeria syndrome (HGPS), are caused by single point mutations. HGPS is a rare disorder that causes premature aging and is usually caused by a de novo point mutation in the LMNA gene. Base editors (BEs) composed of a cytidine deaminase fused to CRISPR/Cas9 nickase are highly efficient at inducing C to T base conversions in a programmable manner and can be used to generate animal disease models with single amino-acid substitutions. Here, we generated the first HGPS monkey model by delivering a BE mRNA and guide RNA (gRNA) targeting the LMNA gene via microinjection into monkey zygotes. Five out of six newborn monkeys carried the mutation specifically at the target site. HGPS monkeys expressed the toxic form of lamin A, progerin, and recapitulated the typical HGPS phenotypes including growth retardation, bone alterations, and vascular abnormalities. Thus, this monkey model genetically and clinically mimics HGPS in humans, demonstrating that the BE system can efficiently and accurately generate patient-specific disease models in non-human primates.
Animals ; Disease Models, Animal ; Female ; Gene Editing ; Humans ; Lamin Type A/metabolism* ; Macaca fascicularis ; Progeria/pathology*