Objective:To explore the effects of long noncoding RNA adenosine diphosphate-dependent glucokinase antisense RNA 1(ADPGK-AS1) on the proliferation, migration and invasion of human retinoblastoma (RB) Y-79 cells and its regulatory effect on microRNA-623 (miR-623).Methods:The peritumoral tissue and RB specimens were collected from 39 eyes of 39 patients with RB during surgery in The First Affiliated Hospital of Zhengzhou University and Zhumadian Central Hospital from February 2017 to November 2018.Real-time fluorescence quantitative PCR was employed to detect the expression of ADPGK-AS1 and miR-623 in the specimens.Human RB line Y-79 cells were cultured in vitro and divided into small interfering RNA-normal control (siRNA-NC) group, siRNA-ADPGK-AS1 group, microRNA (miR)-NC group, miR-623 group, siRNA-ADPGK-AS1+ anti-miR-NC group and siRNA-ADPGK-AS1+ anti-miR-623 group.The cell proliferation rate was detected by MTT method.Transwell cell experiment was performed to detect the number of migrating and invading cells.The dual luciferase reporter experiment was used to evaluate the targeting relationship between ADPGK-AS1 and miR-623.The expression of Ki-67, matrix metalloproteinases (MMP)-2, and MMP-9 in the cells was detected by Western blot assay.Written informed consent was obtained from each patient prior to any medical examination and treatment.This study protocol adhered to the Declaration of Helsinki.The use of the human specimens was approved by an Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2017-KY-73). Results:Compared with the peritumoral tissue, the relative expression level of ADPGK-AS1 in the RB tissue was significantly increased, and the relative expression level of miR-623 was significantly reduced ( t=40.522, 48.497; both at P<0.01). Compared with the siRNA-NC group, both the relative expression level of Ki-67 protein and the proliferation A value of RB Y-79 cells were significantly reduced in the siRNA-ADPGK-AS1 group ( t=26.833, 18.522; both at P<0.01). The relative expression levels of MMP-2 and MMP-9 proteins in the siRNA-ADPGK-AS1 group were significantly lower than those in the siRNA-NC group ( t=22.123, 26.183; both at P<0.01). The number of migrating and invading cells in the siRNA-ADPGK-AS1 group was significantly less than that in the siRNA-NC group ( t=12.385, 19.201; both at P<0.01). The dual luciferase report experiment confirmed that ADPGK-AS1 targeted miR-623.The protein expression levels of the Ki-67, MMP-2 and MMP-9 in the miR-623 group were significantly lower than those in the miR-NC group ( t=22.137, 22.200, 21.094; all at P<0.01). Compared with the miR-NC group, the proliferation A value of Y-79 cells in the miR-623 group was significantly lower, and the number of migrating and invadoing cells was significantly less ( t=16.398, 11.400, 17.846; all at P<0.01). The relative expressions levels of Ki-67, MMP-2 and MMP-9 proteins in the siRNA-ADPGK-AS1+ anti-miR-623 group were significantly higher than those in the siRNA-ADPGK-AS1+ anti-miR-NC group ( t=20.795, 17.493, 23.479; all at P<0.01). Compared with the siRNA-ADPGK-AS1+ anti-miR-NC group, the proliferation A value of Y-79 cells in the siRNA-ADPGK-AS1+ anti-miR-623 group was significantly increased ( t=15.600, P<0.01), and the number of migrating and invading cells was obviously elevated ( t=14.495, 17.855; both at P<0.01). Conclusions:Knockdown of ADPGK-AS1 gene can inhibit the proliferation, migration and invasion of Y-79 cells by up-regulating the expression of miR-623.

Objective:Stephania tetrandra is widely used in traditional Chinese medication.In present study,we studied the resource status of it in Anhui and Jiangxi Province.Methods:We inspected relevant documents to confirm which counties we would investigate;and then,we interviewed many relative local people and carried out sample-plot survey.Results:The resource of wild Stephania tetrandra in the two provinces were both seriously damaged.Conclusion:Large-scale promotion of artificial cultivation was required to ensure the supply and to realize the reasonable and sustainable utility of this medicinal material.

Background Orbital cavernous hemangiomas (OCH) is a common benign orbital tumor in adult,and accurate localization and diagnosis before operation supports the significant premise for surgical safety and success of tumor extraction.Objective This study was to research the clinical characteristics,preoperative diagnosis,the selection for different surgical approaches,therapeutic effectiveness and complication prevention of OCH.Methods The clinical data of 117 eyes of 117 patients with OCH who received surgery were retrospectively analyzed.The patients received surgery in Henan Eye Institute,Henan Eye Hospital from January 2011 to December 2014 and followed-up for 3 months to 5 years.The visual acuity,exophthalmos,ocular movement,orbital A/B ultrasound,color Doppler image,CT and MRI were examined before and after surgery.Results The primary clinical manifestations of OCH were gradual exophthalmos and impaired vision.The accordance rate of preoperative diagnosis and pathological diagnosis was 100％ in the group of patients.The surgical approachs included conjunctival approach in 52.14％ (61/117),lateral orbitectomy in 30.77％ (36/117),anterior approach in 16.24％ (19/117) and lateral combined with medial approach in 0.85％ (1/117).At the end of followed-up,visual acuity was significantly improved in 30.77％ (36/117),declined in 8.55％ (10/117) and unchanged in 60.68％ (71/117).Temporary complications after surgery were pupil dilatation in 14.53％ (17/117),emorrhoea in 1.71％ (2/117),ocular motility disorders in 16.24％ (19/117) and ptosis in 4.27％ (5/117).The permanent complications after operation were pupil dilatation in 2.56％ (3/117),visual loss in 0.85％ (1/117) and permanent abduction imitation in 0.85％ (1/117).Conclusions Accurate qualitative and site-specific diagnosis and correct choice of surgeries for OCH depend on clinical and iconographical examinations.Suitable surgical approach and operative skill are helpful to the therapeutic outcome and safety of OCH.

OBJECTIVETo clone and sequence the open reading frame of beta-amyrin synthase (bAS) from Glycyrrhiza uralensis.

METHODThe primers were designed according to the cDNA sequence of beta-amyrin synthase from G. glabra reported by Hiroaki HAYASHI, and the open reading frame of beta-amyrin synthase was cloned by RT-PCR strategy with the template of total RNA extracted from roots of G. uralensis.

RESULTThe GubAS (GenBank Accession number: FJ627179) was 2 289 bp in length encoding one pelypeptide of 762 amino acid. Deduced amino acid sequence had 99%, 92%, 90%, 90% and 89% homology to the amino acid sequence of G. glabra, Lotus japonicus, Pisum sativum, Medicago truncatula, Glycine max, respectively.

CONCLUSIONThe open reading frame of bAS from G. uralensis is cloned and reported for the first time. The conclusion will provide a foundation for exploring the mechanism of triterpenes biosynthesis.

Cloning, Molecular ; Glycyrrhiza uralensis ; classification ; enzymology ; genetics ; Intramolecular Transferases ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants ; classification ; genetics

OBJECTIVETo analyze the correlation between content of glycyrrhizic acid and the single nucleotide polymorphism of beta-amyrin synthase (bAS) in Glycyrrhiza uralensis.

METHODglycyrrhizic acid content in 80 samples of the cultivated G. uralensis were determined by HPLC; According to the very significant level (P < 0.000 1), 80 samples in accordance with glycyrrhizic acid will be grouped by SAS 9.0; Using RT-PCR strategy to amplification the Open Reading Frame of beta-amyrin synthase with the template of total RNA extracted from roots of G. uralensis and then using DNAman to analyze the relationship between glycyrrhizic acid content and the single nucleotide polymorphism of beta-amyrin synthase (bAS).

RESULTThere exited two mutation sites 94 bp and 254 bp, G/A conversion occurred at 94 bp site, which belonged to a missense mutation. G/A conversion led to the corresponding amino acid conversion (Gly --> Asp); C/T conversion occurred at 254 bp site, which belonged to a synonymous mutation. According to sequence variation, the samples were divided into four genotypes: G-T genotype, A-T genotype, G/A-C genotype and G-T genotype.

CONCLUSIONA-T genotype, G/A-C genotype and G-T genotype are correlated with the high content of glycyrrhizic acid.

Genotype ; Glycyrrhiza uralensis ; enzymology ; genetics ; metabolism ; Glycyrrhizic Acid ; metabolism ; Intramolecular Transferases ; genetics ; Polymorphism, Single Nucleotide ; Reproducibility of Results

This study was aimed to assess the agreement between pulse pulmonary arterial oxygen saturation (StO2) by oximetry in trachea and the mixed venous oxygen saturation (SvO2) by fiberoptic pulmonary arterial catheter, and to evaluate the accuracy of StO2 monitoring during hypoxia. 10 mongred dog's were used. After anesthesia was induced and thorax was opened, we placed a fiberoptic pulmonary artery catheter directly through the dog's right ventricular outlet. Then, we placed and adjusted the trachea catheter, attached the oximetry probe to trachea carina till the high quality StO2 PPG signal was obtained, and till the readings were stable at about +/-2% from fiberoptic catheter. The pair readings of StO2 and SvO2 were recorded at every minute interval in 10 minutes when circulation was stable. Decreasing the inhaled oxygen concentration till the tongue's SpO2 decreased to 60%; recording the changes of StO2 and SvO2 in every 5% drop of tongue's SpO2 when the tongue's SpO2 decreased from 100% to 50%. The results showed that there was a good agreement between the two methods for pulmonary arterial oxygen saturation measurement. However, the difference between the two methods was great and unacceptable during the presence of hypoxia.
Animals ; Catheterization, Swan-Ganz ; methods ; Dogs ; Female ; Hypoxia ; blood ; Male ; Monitoring, Physiologic ; methods ; Oximetry ; instrumentation ; methods ; Oxygen ; blood ; Pulmonary Artery ; Trachea

OBJECTIVETo analyze heterologous expression in Saccharomyces cerevisiae of two genotypes: beta-AS (A-T) genotype which is related to high content of glycyrrhizic acid and beta-AS(G-C) genotype which is related to low content of glycyrrhizic acid, and compare two different genotypes on the impact of beta-amyrin production in order to provide a foundation for licorice molecular breeding.

METHODThe 2 289 bp fragment in plasmid pMD-19T encoding beta-amyrin synthase was subcloned into the yeast-Escherichia coli shuttle vector pY26, thus an expression recombinant plasmid PY-beta-AS containing target gene was constructed. The PY-beta-AS was introduced into defective mutant INVSc1 of S. cerevisiae by LiAc method, after induced by IPTG, the content of beta-amyrin was determined by GC-MS.

RESULTGC-MS analysis demonstrates that the an occurring peak corresponding to beta-amyrin standards was detected with the same retention time, which is absent in the cell transform with empty vector. Results showed the peak was beta-amyrin and the percentage of beta-amyrin in two genotypes: beta-AS (A-T) genotype and beta-AS (G-C) genotype were 19.08% and 1.40%, respectively. Thus the beta-amyrin synthase exhibited the activity of catalyzing 2, 3-oxidosqualene to beta-amyrin.

CONCLUSIONThe catalytic efficiency of beta-AS(A-T) genotype is higher than that of beta-AS(G-C) genotype, which can lay the foundation for licorice molecular breeding.

Catalysis ; Cloning, Molecular ; Genotype ; Glycyrrhiza uralensis ; chemistry ; enzymology ; genetics ; Intramolecular Transferases ; chemistry ; genetics ; metabolism ; Plant Proteins ; chemistry ; genetics ; metabolism ; Polymorphism, Genetic ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics

Objective:To investigate the clinical features and prognosis of male with anti-melanoma differentiation-associated gene 5 (MDA5) autoantibody.Methods:The clinical data of 246 patients with DM and anti-MDA5 autoantibodies hospitalized by Jiangsu Myositis Cooperation Group from 2017 to 2020 were collected and retrospectively analyzed. Chi-square test was performed to compared between counting data groups; Quantitative data were expressed by M ( Q1, Q3), and rank sum test was used for comparison between groups; Single factor survival analysis was performed by Kaplan-Meier method and Log rank test; Cox regression analysis were used for multivariate survival analysis. Results:①The male group had a higher proportion of rash at the sun exposure area [67.1%(47/70) vs 52.8%(93/176), χ2=4.18, P=0.041] and V-sign [50.0%(35/70) vs 30.7%(54/176), χ2=8.09, P=0.004] than the female group. The male group had higher levels of creatine kinase [112(18, 981)U/L vs 57 (13.6, 1 433)U/L, Z=-3.50, P<0.001] and ferritin [1 500 (166, 32 716)ng/ml vs 569 (18, 14 839)ng/ml, Z=-5.85, P<0.001] than the female group. The proportion of ILD [40.0%(28/70) vs 59.7%(105/176), χ2=7.82, P=0.020] patients and the red blood cell sedimentation rate[31.0(4.0, 101.5)mm/1 h vs 43.4(5.0, 126.5)mm/1 h, Z=-2.22, P=0.026] in the male group was lower than that of the female group, but the proportion of rapidly progressive interstitial lung disease (PR-ILD) [47.1%(33/70) vs 31.3%(55/176), χ2=5.51, P=0.019] was higher than that of the female group. ②In male patients with positive anti-MDA5 antibodies,the death group had a shorter course of disease[1.0(1.0, 3.0) month vs 2.5(0.5,84) month, Z=-3.07, P=0.002], the incidence of arthritis [16.7%(4/24) vs 42.2%(19/45), χ2=4.60, P=0.032] were low than those in survival group,while aspartate aminotransferase (AST)[64(22.1, 565)U/L vs 51(14,601)U/L, Z=-2.42, P=0.016], lactate dehydrogenase (LDH) [485(24,1 464)U/L vs 352(170, 1 213)U/L, Z=-3.38, P=0.001], C-reactive protein (CRP) [11.6(2.9, 61.7) mg/L vs 4.95(0.6, 86.4) mg/L, Z=-1.96, P=0.050], and ferritin levels [2 000(681, 7 676) vs 1 125 (166, 32 716)ng/ml, Z=-3.18, P=0.001] were higher than those in the survival group, and RP-ILD [95.8%(23/24) vs 22.2%(10/45), χ2=33.99, P<0.001] occurred at a significantly higher rate. ③Cox regression analysis indicated that the course of disease LDH level, and RP-ILD were related factors for the prognosis of male anti-MDA5 antibodies [ HR (95% CI)=0.203(0.077, 0.534), P=0.001; HR (95% CI)=1.002(1.001, 1.004), P=0.003; HR (95% CI)=95.674 (10.872, 841.904), P<0.001]. Conclusion:The clinical manifestations of male anti-MDA5 antibody-positive patients are different from those of female. The incidence of ILD is low, but the proportion of PR-ILD is high. The course of disease, serum LDH level, and RP-ILD are prognostic factors of male anti-MDA5 antibody-positive patients.

Objective To investigate the current situation in Chinese rheumatologic physicians' clinical diagnosis and evaluation of Takayasu's arteritis (TA).Methods Nineteen rheumatology experts and three vascular surgery specialists in China were invited to make the nationwide investigation for the first time about the diagnosis and disease activity evaluation of TA in China,through the questionnaire survey on the internet.Weighted average was used to calculate the average scores of corresponding problems.Results Chinese experts mainly adopted 1990 American College of Rheumatology (ACR) classification criteria for clinical diagnosis of TA.In details,symptoms of age,limb claudication and amaurosis,signs including pulselessness or pulse weakening,vascular bruits,increasing bilateral pulse pressure and hypertension and acute phase reactants (APR) were critical to the clinical diagnosis of TA.Besides,noninvasive imaging examinations,such as computed tomography angiography (CTA),magnetic resonance angiography (MRA),vascular ultrasonography,and positron emission tomography (PET) were also of great importance.In the aspect of disease activity assessment,Chinese experts mainly used Kerr scoring tool.APR and noninvasive radiological examinations were considered with vital value.Some TA patients with carotid artery involvement were recommended using vascular ultrasonography,while others with pulmonary artery and thoracic/abdominal aorta trunk involvement were preferred CTA other than MRA.Conclusions APR and noninvasive imaging examinations were thought with great help to make clinical diagnosis and evaluation of TA for Chinese physicians.

Objective: To analyze the diagnosis and treatment of acute myeloid leukemia (AML) complicated with myeloid sarcoma (MS). Methods: The clinical features, diagnosis and therapy of 3 AML who were treated with routine regimen induction and consolidation therapy in Shanghai Beizhan Hospital and showed MS during the marrow remission stage were retrospectively analyzed. Results: Three female patients included 2 cases of CBFβ-MYH11 positive and 1 case of AML1-ETO positive. One patient occurred in the first complete remission(CR₁) and 2 patients occurred in CR₂. The involved sites included brain, vagina, and breast respectively. Three patients all received short efficacy after chemotherapy, including 1 patient in bone marrow remission stage, 1 patient died of after bone marrow relapse and 1 patient without follow-up. Conclusions For AML patients in remission with extramedullary lesions, pathological examination is warranted to confirm the diagnosis of the disease. Systemic chemotherapy is still the main treatment for MS. Central nervous system(CNS) leukemia treatment should be performed to the patients with CNS involved. Allogeneic hematopoietic stem cell transplantation may be the best option for the treatment of these patients.