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Objective:To explore and validate the targets of Salvia miltiorrhiza in myopia using network pharmacology and molecular docking technology. Methods:The TCMSP database was used to extract the targets of Salvia miltiorrhiza.GeneCards, DisGeNET, Malacard and OMIM databases were used to extract the myopia-related targets.The target intersection was taken, and the intersecting targets were selected to extract the corresponding active ingredients of traditional Chinese medicine (TCM) and construct the pharmacological regulatory network of TCM using Cytoscape.The protein interaction network map for the key target genes was constructed using the String database, and the relevant proteins were selected to download the three-dimensional structures of the active ingredients from the PubChem database, and molecular docking was performed using AutoDockvina software.Twelve 3-week-old guinea pigs were induced with lens-induced myopia (LIM) in the right eye and randomly divided into normal saline group and sodium danshensu group, with 6 animals in each group.During the maintenance of LIM, periocular injection of 1 ml normal saline or sodium danshensu was performed daily.The contralateral eye was used as a negative control.On days 0, 14, and 28 of the experiment, the axial length of both eyes was measured by A-scan ultrasonography, and the refractive status was assessed with a streak retinoscope.To avoid individual differences, relative spherical equivalent (treated eye-contralateral eye) and relative axial length (treated eye-contralateral eye) were compared.On day 28, the relative expression levels of hypoxia-inducible factor-1α (HIF-1α) and transforming growth factor-β1 (TGF-β1) proteins were determined by Western blot.The feeding and use of laboratory animals followed the 3R principle, and the research program was approved by the Ethics Committee of Experimental Animal Center of Zhengzhou University (No.ZZU-LAC 202320405[02]). Results:Sixteen intersecting key targets were screened for myopia and TCM components derived from Salvia divinorum.A TCM network pharmacology map and protein interaction map were constructed with Salvia divinorum as a drug candidate, and the corresponding proteins of target genes, such as MMP2, TGFB1, and MMP9 were screened to perform molecular docking with the active ingredients, such as lignocellulosic acid, danshensu, tanshinone ⅡA, and so on.After 14 days of induction, the relative spherical equivalent and relative axial length were (-4.67±1.03)D and (0.67±0.26)mm in sodium danshensu group, and (-6.30±1.22)D and (1.08±0.34)mm normal saline group, indicating slower myopia progression and axial elongation in sodium danshensu group, and the differences were statistically significant ( t=2.412, P=0.039; t=2.750, P=0.049). The relative expression levels of HIF-1α protein were 0.20±0.01, 1.29±0.05 and 0.63±0.02, and the relative expression levels of TGF-β1 protein were 0.93±0.05, 0.25±0.01 and 0.74±0.05 in the negative control, normal saline and sodium danshensu groups, respectively.The expression of HIF-1α protein was higher in sodium danshensu group than in negative control group but lower than in the normal saline group, and the expression of TGF-β1 protein was lower in sodium danshensu group than in negative control group but higher than in the normal saline group, showing statistically significant differences (all at P<0.05). Conclusions:Natural compounds extracted from Salvia divinorum extracts may serve as potential drug candidates to combat scleral hypoxia and improve scleral extracellular matrix remodeling.
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Objective To screen the active components of total flavonoid extracts of Sarcandra glabra to promote megakaryocyte differentiation.Methods(1)A model of megakaryocyte differentiation disorder was established by co-culturing human megakaryocytic leukaemia cells(Dami)with human bone marrow stromal cells(HS-5)as an evaluation system,and the experimental groupings were as follows:the Dami group(Dami),the control group(Dami+HS-5),and the PMA group[Dami+HS-5+5 ng·mL-1 foprolol 12-tetradecanoate 13-acetate(PMA)],and model group[Dami+HS-5+1%rabbit anti-rat platelet serum(APS)+5 ng·mL-1 PMA]were cultured for 48 hours.The expressions of megakaryocyte differentiation and maturation surface marker molecules,CD41a and CD61 were detected by flow cytometry.(2)Forty-nine SD male rats were randomly divided into blank plasma group,15-minute group,30-minute group,60-minute group,90-minute group,120-minute group,and 240-minute group,with 7 rats in each group.The rats in each administration group were gavaged with 1.26 g·kg-1 of total flavonoids extracts of Sarcandra glabra,and blood was collected at six set time points(15,30,60,90,120,240 minutes)for the preparation of time-dependent serum-containing plasma of total flavonoids extracts of Sarcandra glabra.(3)Ultra-high performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UHPLC-Q-TOF/MS)was used to analyze the plasma of the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra,and the peak area was used to construct a matrix(X-matrix)of the amount of chemical composition change over time in the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra.The collected time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra at six different time points was used to intervene in the model of megakaryocyte differentiation and maturation disorder,and the expression of cell surface molecules CD41a and CD61 was detected by flow cytometry to construct the matrix of effect of time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra(Y-matrix).(4)After the data of X and Y matrices were standardized,partial least squares(PLS)was used to calculate and analyze the quantitative and qualitative effect relationship,and variable importance for projection(VIP)>1 was used as the threshold to screen the effect components related to the changes of cell surface molecules CD41a and CD61,and chemical composition identification,as the potential effector components in the total flavonoid extracts of Sarcandra glabra were used to promote the differentiation of megakaryocytes,and finally the regression evaluation system was used to verify the efficacy of its medicinal effect.Results(1)Compared with the Dami group,the expression level of CD41a on the surface of Dami cells in the control group was significantly increased(P＜0.05).Compared with the control group,the expression levels of CD41a and CD61 on the surface of Dami cells in the PMA group were significantly increased(P＜0.01).Compared with the PMA group,the expression levels of CD41a and CD61 on the surface of Dami cells in the model group were significantly reduced(P＜0.01).(2)Compared with the blank plasma group,the expression levels of the molecules CD41a and CD61 on the surface of Dami cells at each time point of 15,30,60,90,120,and 240 minutes were significantly increased(P＜0.01),and the expression levels of CD41a and CD61 were both highest in the 30-minute group.The potential effective components with VIP value greater than 1 were screened out in the positive and negative ion mode,and 540.3638@12.25 and 559.2991@11.53 were selected for pharmacodynamic verification.559.2991@11.53 was identified as daucosterol(Dau),540.3638@12.25 was identified as rosmarinic acid 4-O-β-D-glucoside(Ros).After Ros and Dau intervened in the megakaryocyte differentiation and maturation disorder model respectively,the expression levels of CD41a and CD61 on the surface of Dami cells in the low-,medium-and high-dose groups(40,60 and 80 μg·mL-1)of Ros and Dau were significantly increased compared with the model group(P＜0.05,P＜0.01).Conclusion Ros and Dau may be the active components of the total flavonoids extracts of Sarcandra glabra to promote the differentiation of megakaryocytes.
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Objective:To identify the causative gene and observe the phenotypic characteristics of a family with isolated microphthalmia-anophthalmia-coloboma (MAC).Methods:A retrospective clinical study. One patient (proband) and 3 family members of a family with MAC visited the Henan Eye Hospital from May 2019 to May 2022 were included in the study. The patient's medical history and family history were inquired in detail, and the best corrected visual acuity (BCVA), slit lamp microscope, fundus photography, optical coherence tomography (OCT), ophthalmological B mode ultrasound and axial length (AL) measurement were performed. The peripheral venous blood of the proband, his parents and brother was collected for Trio whole-exome sequencing and pathogenic gene screening. Fluorescence quantitative Polymerase chain reaction was used to verify the suspicious variations. The clinical features of the patient's ocular and systemic also were observed.Results:The proband, male, was 3 years old at the first visit. The horizontal pendular nystagmus was detected in both eyes. Vertical elliptical microcornea and keyhole-shaped iris colobomas were detected in both eyes. The objective refraction at first visit (3 years old) was -4.00 DS/-0.50 DC×105° (OD) and -3.50 DS/-1.25 DC×80° (OS). Refraction and BCVA at 6 years old: -6.50 DS/-2.00 DC×110°→0.05 (OD) and -6.00 DS/-1.50 DC×80°→0.2 (OS). The AL at 4 years and 10 months old was 24.62 mm (OD) and 23.92 mm (OS), respectively. The AL at 5 years and 7 months old was 25.24 mm (OD) and 24.36 mm (OS), respectively. Ultrasonography shows tissue defects in both eyes. Fundus photography showed the inferior choroidal coloboma involving optic disc. OCT showed the optic disc in both eyes was abnormal with colobomas around, and the retinal neurosensory layer in colobomas area was disordered and thin; the retinoschisis was visible in the left eye. The proband's parents and siblings have normal phenotypes. Whole exome sequencing reveals a denovo heterozygous deletion of YAP1 gene: YAP1, chr11: 10280247-102100671, NM_ 001130145, loss 1 (EXON: 6-9). The results of bioinformatics analysis were pathogenic variants. Parents and siblings were of the wild type. Conclusions:Loss of heterozygosity in exons 6-9 of YAP1 gene is the pathogenic variation in this family. It can cause abnormal development of anterior segment, chorioretinal colobomas, deepening of axial myopia, even severe macular colobomas and retinoschisis.
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Objective:To identify the causative genes of the posterior microphthalmia-retinal pigment degeneration family.Methods:A retrospective clinical study. One child (proband) and 3 family members of a family with posterior microphthalmia-retinitis pigmentosa diagnosed by clinical and genetic examination at Henan Provincial People's Hospital in July 2019 were included in the study. Medical history and family history, and draw pedigree of the patients was collected. Visual acuity, visual field, fundus color photography, optical coherence tomography and electroretinogram (ERG) were examined. The peripheral venous blood of the proband, his parents and sister, and extract the whole genome DNA was collected. Whole-exome sequencing was used to detect genetic variations, the suspected pathogenic variations were verified by Sanger sequencing, and the pathogenicity was determined by bioinformatics analysis.Results:The parents discovered the proband was poor vision at the age of 10 months. At the age of 3, the best corrected visual acuity of the right eye and the left eye were 0.3 and 0.4, respectively. No abnormality was found in anterior segment. Extremely high hyperopia in both eyes. The axial length was 14.47 mm and 15.78 mm, respectively. The optic disc of both eyes was relatively small and flushed, retinal folds can be observed in macular area, and no obvious pigment deposition was found. ERG examination showed that the rod system response and the maximal combined response of both eyes decreased slightly to moderately, and the single-flash cone response and the 30 Hz flicker response decreased moderately to severely. Genetic analysis revealed two novel mutations in the membrane frizzled-related protein ( MFRP) gene in the proband: c.363delC/p.Thr121Thrfs*16, c.1627C>T/p.Gln543Stop,37 in exon 4 and 13, the former was a frameshift mutation, encoding 16 amino acids and then terminated, and the latter was an nonsense mutation, truncated 37 amino acids, both which were predicted to be pathogenic and segregate with disease. The mother and sister carried c.363delC, and the father carried c.1627C>T. Conclusion:MFRP gene c.363delC/p.Thr121Thrfs*16, c.1627C >T/p.Gln543Stop, 37 compound heterozygous mutation may be the pathogenic gene of this family.
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Objective:To investigate the feasibility of ultrasound-guided percutaneous endomyocardial septal cryoablation of in vitro porcine hearts and to compare its effect with the percutaneous endomyocardial radiofrequency ablation.Methods:Experiment 1: Six in vitro porcine hearts were divided into 1 min ( n=2), 3 min ( n=2) and 5 min ( n=2) groups according to the cryoablation time, and all were subjected to ultrasound-guided percutaneous intra-myocardial septal cryoablation at 100% power respectively. After cryoablation, ultrasound images, the size of the solid dissection of the ice ball, and the size of the necrotic area after melting of the frozen ice ball were measured. Experiment 2: The in vitro porcine hearts were divided into cryoablation group ( n=3) and radiofrequency ablation group ( n=3), and ultrasound-guided percutaneous endomyocardial septal cryoablation and radiofrequency ablation were performed with 100% cryo power and 40 W radiofrequency power, and the extent of complete necrotic area and incomplete necrotic area were compared between the two ablation methods after 1 min. Results:Experiment 1: In the 1 min cryoablation time group ( n=2), the short diameter of the puck measured by ultrasound was (8.00±0.84)mm, the short diameter of the puck measured by solid was (8.38±1.19)mm, and the short diameter of the necrotic zone measured by solid was (8.35±0.83)mm; in the 3 min group ( n=2), the short diameter of the puck measured by ultrasound was (19.4±0.28)mm, and the short diameter of the puck measured by solid was (19.03±0.33)mm, solid measurement of the short diameter of the necrotic zone was (19.16±0.25)mm; in the 5 min group ( n=2), the short diameter of the puck measured under ultrasound was (26.4±2.54)mm, solid measurement of the short diameter of the puck was (26.01±0.24)mm, and solid measurement of the short diameter of the necrotic zone was (24.82±0.25)mm. Randomized blocks analysis of variance was performed on this data and the difference of block Factor b (freezing time: 1 min, 3 min, 5 min) among the three groups was statistically significant( F=505.884, P<0.001). The SNK- q test showed that all three groups differed from each other(all P<0.05). The analysis results for the treatment factors K (measurement modality-ultrasound image measurements, solid anatomical measurements of the puck, and measurements of the necrotic area after melting of the frozen puck) was not statistically significant ( F=0.470, P=0.635). Experiment 2: In the RF ablation group ( n=3), the ratio of incomplete necrotic zone to the radius of the RF ablation area was 0.64±0.01; in the cryoablation group ( n=3), the ratio of incomplete necrotic zone to the radius of the ablation area was 0.26±0.02. The difference was statistically significant( P=0.002) and it can be considered that the incomplete necrotic zone of cryoablation was smaller than that of RF ablation. Conclusions:Percutameous intramyocardial septal cryoablation is controllable in scope, ultrasound image evaluation of ablation area is more accurate and incomplete necrosis area is small, which may have potential applications in cardiac ablation.
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Objective:To quantitatively evaluate myocardial microcirculation perfusion changes after percutaneous intracardiac septal radiofrequency ablation (PIMSRA, Liwen procedure) in patients with obstructive hypertrophic cardiomyopathy (HOCM) using myocardial contrast echocardiography (MCE) and to further establish the effect of the procedure and predict the long-term prognosis of patients.Methods:A total of 45 patients with HOCM treated by the Liwen procedure in the First Affiliated Hospital of Air Force Medical University (Xijing Hospital) from July 2019 to June 2020 were included. MCE was performed before and 6 months after surgery, respectively. Time-intensity perfusion curve analysis was performed using QLab 10.8 offline software to obtain quantitative parameters of myocardial microcirculation perfusion including myocardial blood volume (A value), myocardial blood flow velocity (β value), and myocardial blood flow (A×β value), then the changes in parameters before and after the procedure were compared.Results:After the Liwen procedure, the mean septal thickness and mean left ventricular free wall thickness were significantly reduced, accompanied by a significant reduction in the left ventricular outflow tract pressure gradient (LVOT-PG) and mitral regurgitation length, and a significant improvement in mitral systolic antegrade motion (SAM) (all P<0.001). In addition, the left ventricular ejection fraction (LVEF) did not deteriorate significantly ( P=0.560) and the E/e′ ratio decreased after the procedure ( P=0.015). Besides, the A values of both the ventricular septum and the left ventricular free wall were not significantly changed compared to those before procedure ( P>0.05), whereas the β values and A×β values were obviously increased ( P<0.05). Conclusions:Myocardial blood velocity and myocardial blood flow in the septum and left ventricular free wall were significantly increased in patients with HOCM after the Liwen procedure, suggesting a significant improvement in myocardial microcirculation perfusion. MCE provides a non-invasive quantitative evaluation parameters of myocardial microcirculation perfusion for the Liwen procedure for the treatment of HOCM.
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Objective To observe the clinical effect of minimally invasive vitreoretinal (MIV) surgery combined with a modified suprachoroidal drainage surgery for retinal detachment associated with choroidal detachment (RRDCD).Methods A prospective clinical study.A total of 27 patients (27eyes) diagnosed as RRDCD were recruited in this study.There were 16 males and 11 females,with an average of (53.67± 14.82) years.The mean intraocular pressure (IOP) was (8.2± 2.1) mmHg (1 mmHg=0.133 kPa) and best corrected visual acuity (BCVA) of minimum resolution angle logarithm (logMAR) was 1.87±0.58.All subjects underwent 23G MIV combined a modified suprachoroidal drainage surgery,which 23G stab knife and 1 ml syringe needle were used for surgery.The visual outcome,IOP,rate of retinal reattachment and complications were comparatively analyzed preoperatively and postoperatively.Results At 1 day,10 days,1 month and 3 months after surgery,the average of logMAR BCVA were 1.62 ± 0.67,1.51 ± 0.63,1.39 ± 0.54,1.32± 0.56 and the mean of IOP were (13.47 ± 5.06),(14.43 ± 4.09),(14.89 ± 4.30),(15.38 ± 3.37) mmHg,respectively.There were significant differences of logMAR BCVA and IOP between before and after surgery (F=6.19,15.21;P＜0.05).Retinal reattachments were achieved in 27 eyes (100％) at 1 day and 10 days after surgery.At 1 month and 3 months after surgery,the rate of retinal reattachment were 88.89％ (24 eyes) and 85.19％ (23 eyes),respectively.No severe complications such as endophthalmitis and choroidal hemorrhage were found at follow-up visits.Conclusion MIV combined with a modified suprachoroidal drainage surgery is an effective and safe treatment for RRDCD,which can promote retina tear closure,improve visual acuity.
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Objective To analyze the pathogenic gene types and phenotypic characteristics of 6 albinism families.Methods A retrospective series of case studies.Six probands of albinism and 20 family members were recruited for this study,5 probands with clinical manifestations of oculocutaneous albinism (OCA) and 1 proband of ocular albinism (OA).Genomic DNA was extracted from peripheral venous blood which was collected from 6 probands and 20 family members.Genetic variations were screened by whole-exome sequencing or Sanger sequencing and then analyzed the relationship between genotypes and phenotypes.Results Genetic sequencing identified 6 potential pathogenic variants in 4 probands,including 2 compound heterozygous mutations in the 2 genes [TYR (c.1037-7T＞A,c.925_c.926insC),OCA2 (c.2359G＞A,c.587T＞C)] associated with OCA1 and OCA2,and 2 hemizygous mutations in the GPR143[GPR143 (c.11C ＞ G),GPR 143 (c.333 G ＞ A)] as sociated with OA 1,respectively.In which,5 were novel mutations and confirmed by Sanger sequencing.One case was accorded with OCA in clinical phenotype,but genetic diagnosis was OA1,the others were agreement between clinical diagnosis and genetic diagnosis.Conclusion There are 4 families with mutations in 6 families,representative of 3 type of albinism (OCA1,OCA2,OA1).
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To promote clinical medical students' personal harmonious development,we have established the new problem-based teaching mode and reform the evaluation system in clinical teaching of obstetrics and gynecology and have obtained more satisfied teaching result.
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Medicine is a practical subject,which requires medical students to deepen and expand the understanding of the theory by clinical training,and establish clinical thinking.In our noviciate reform of obstetrics and gynecology,we have established the new problem-based teaching mode,which is an effective method in fostering the ability of information collection,innovation and practice,and have obtained more satisfactory results.