Sucrose synthase plays a crucial role in the plant sugar metabolism pathway by catalyzing the production of uridine diphosphate (UDP)-glucose, which serves as a bioactive glycosyl donor for various metabolic processes. In this study, a sucrose synthase gene named CtSus was cloned from Cistanche tubulosa, a traditional Chinese medicine known for its rich content of diverse glycosides, based on transcriptome analysis results. The open reading frame of CtSus was 2 418 bp long encoding 805 amino acids. Sequence analysis revealed conserved domains associated with the plant sucrose synthase family at both the N-terminal and C-terminal regions of CtSus protein. Phylogenetic analysis demonstrated that CtSus shares a close evolutionary relationship with sucrose synthases from other plants within the same order. Functional identification of CtSus was performed through whole-cell catalysis coupling with UGT71BD1, a characterized glycosyltransferase enzyme. The results showed that the introduction of CtSus significantly enhanced the conversion rate of glycosylation catalyzed by UGT71BD1. The soluble expression of CtSus in Escherichia coli was further achieved using the pCold^TM TF expression vector. In vitro enzymatic assay indicated the activity of CtSus to catalyze the formation of UDP-glucose in the presence of sucrose and UDP. Real-time quantitative-polymerase chain reaction results showed that the expression patterns of CtSus gene in different parts of C. tubulosa and the suspension cell cultures of C. tubulosa under drought stress correlated with the accumulation patterns observed for phenylethanol glycosides, respectively. Furthermore, key amino acids and their interactions between enzyme and substrate were explored based on protein structure prediction and molecular docking results. Overall, our findings identify a sucrose synthase CtSus responsible for the supply of the active glycosyl donor UDP-glucose during the biosynthesis of glycoside products in C. tubulosa and have also provided a gene element that can be utilized in engineering strain construction for glycoside products production.

Objective: To analyze the clinical and genetic characteristics of pediatric patients with dual genetic diagnoses (DGD). Methods: Clinical and genetic data of pediatric patients with DGD from January 2021 to February 2022 in Peking University First Hospital were collected and analyzed retrospectively. Results: Among the 9 children, 6 were boys and 3 were girls. The age of last visit or follow-up was 5.0 (2.7,6.8) years. The main clinical manifestations included motor retardation, mental retardation, multiple malformations, and skeletal deformity. Cases 1-4 were all all boys, showed myopathic gait, poor running and jumping, and significantly increased level of serum creatine kinase. Disease-causing variations in Duchenne muscular dystrophy (DMD) gene were confirmed by genetic testing. The 4 children were diagnosed with DMD or Becker muscular dystrophy combined with a second genetic disease, including hypertrophic osteoarthropathy, spinal muscular atrophy, fragile X syndrome, and cerebral cavernous malformations type 3, respectively. Cases 5-9 were clinically and genetically diagnosed as COL9A1 gene-related multiple epiphyseal dysplasia type 6 combined with NF1 gene-related neurofibromatosis type 1, COL6A3 gene-related Bethlem myopathy with WNT1 gene-related osteogenesis imperfecta type XV, Turner syndrome (45, X0/46, XX chimera) with TH gene-related Segawa syndrome, Chromosome 22q11.2 microduplication syndrome with DYNC1H1 gene-related autosomal dominant lower extremity-predominant spinal muscular atrophy-1, and ANKRD11 gene-related KBG syndrome combined with IRF2BPL gene-related neurodevelopmental disorder with regression, abnormal movement, language loss and epilepsy. DMD was the most common, and there were 6 autosomal dominant diseases caused by de novo heterozygous pathogenic variations. Conclusions: Pediatric patients with coexistence of double genetic diagnoses show complex phenotypes. When the clinical manifestations and progression are not fully consistent with the diagnosed rare genetic disease, a second rare genetic disease should be considered, and autosomal dominant diseases caused by de novo heterozygous pathogenic variation should be paid attention to. Trio-based whole-exome sequencing combining a variety of molecular genetic tests would be helpful for precise diagnosis.
Humans ; Abnormalities, Multiple ; Retrospective Studies ; Intellectual Disability/genetics* ; Bone Diseases, Developmental/complications* ; Tooth Abnormalities/complications* ; Facies ; Muscular Dystrophy, Duchenne/complications* ; Muscular Atrophy, Spinal/complications* ; Carrier Proteins ; Nuclear Proteins

Objective To summarize the clinical features of patients with Wilson's disease (WD). Methods A retrospective analysis was performed for the clinical data of 83 patients with WD who were admitted to The Third Hospital of Hebei Medical University from April 2013 to August 2021, including clinical manifestations, Imaging examinations, laboratory examinations, liver histopathological examinations, and ATP7B gene testing results. The patients were divided into groups based on different clinical types. A one-way analysis of variance was used for comparison between groups. Results The youngest age was 3 years for the 83 patients with WD, among whom 39 (46.99%) had an age of ≤18 years, with a mean age of 21.16±14.87 years for all 83 patients. Of all patients, 63.86% had liver-type WD, 31 patients (37.35%) had developed liver cirrhosis at the time of consultation, and 5 patients (6.2%) attended the hospital due to acute or acute-on-chronic liver failure. Of all patients, 62(74.69%) were positive for corneal K-F ring, and the positive rate of K-F ring was 66.04% in the patients with liver-type WD. Among the 83 patients, 79(95.18%) had a reduction in blood ceruloplasmin, and 73(87.95%) had an increase in 24-hour urine copper. The liver histopathological results of 25 patients showed varying degrees of inflammation, fibrosis, steatosis, and copper particle deposition in liver tissue. The ATP7B gene testing results of 25 patients showed that c.2333G > T/p.R778L of exon 8 was the most common mutation site. Conclusion Most patients with WD have the manifestation of liver diseases, and the examinations of corneal K-F ring, serum ceruloplasmin, and 24-h urine copper have their own limitations. Liver pathology and ATP7B gene testing can be performed when it is unable to make a confirmed diagnosis.

Objective:To clarify the clinical efficacy of Yiqi Huoxue recipe in the treatment of liver fibrosis of chronic viral hepatitis.Methods:An open, positive-drug, parallel-controlled study method was applied. A total of 207 cases of liver fibrosis with chronic hepatitis B and C diagnosed with liver biopsy and transient elastography were selected. According to the principle of syndrome differentiation in traditional Chinese medicine, self-made Yiqi Huoxue recipe ( n = 127) and Fuzheng Huayu capsule ( n = 80) were used for the treatment course of 24-48 weeks. Change score of TCM symptom, liver biochemistry, liver stiffness measurement (LSM), and noninvasive liver fibrosis index [aspartate transaminase to platelet ratio index (APRI), and fibrosis-4 score (FIB-4)] were compared between the two groups to evaluate the therapeutic effect of Yiqi Huoxue recipe on liver fibrosis. Results:Yiqi Huoxue recipe group and Fuzheng Huayu capsule group baseline LSM, APRI and FIB-4 was compared, and there was no statistically significant difference between them ( P > 0.05). Yiqi Huoxue recipe and Fuzheng Huayu capsule received patients had improved symptom scores to a certain extent. Hepatic facies, discomfort over liver area, and soreness and weakness of waist and knees ( P < 0.05) was significantly improved in Yiqi Huoxue recipe than Fuzheng Huayu capsule. Liver biochemical indicators (ALT, AST, GGT, ALP) had gradually relapsed with the extension of treatment duration and the normalization rate between the two groups after 24 to 48 weeks had reached 100% vs. 100%, 100% vs. 93.8%, 96.8% vs. 92.3% and 87.5% vs. 81.8%. After 12 weeks of treatment, APRI values ??of both groups had significantly reduced, and after 48 weeks of treatment, LSM values of both groups had significantly improved. Moreover, Yiqi Huoxue recipe FIB-4 score was significantly improved after 48 weeks of treatment, and the difference was statistically significant compared to Fuzheng Huayu capsule group ( P < 0.05). After treatment, LSM, APRI, and FIB-4 total effectiveness in the two groups were 80.0% vs. 63.6%, P = 0.046; 68.4% vs. 52.0%, P = 0.052; 68.4% vs. 62.0%, P = 0.437, respectively. LSM total effectiveness was significantly higher in Yiqi Huoxue recipe treated group than Fuzheng Huayu capsule group. Conclusion:Traditional Chinese medicine Yiqi Huoxue decoction can be used as an optimal treatment for liver fibrosis of chronic viral hepatitis.

Objective: To investigate the safety and efficacy of transcatheter closure of ruptured sinus of Valsava aneurysm(RSVA). Methods: A total of 33 RSVA patients underwent transcatheter closure from January 2006 to March 2017 in our hospital were included in this retrospective study. The RSVA was diagnosed by echocardiography.Different type of occluders were applied for transcatheter closure based on the aortography results. All the patients were followed up after the procedure. Results: The patients were (37.6±12.1) years old,and the male patients accounted for 78.8%(26 cases).RSVA from right coronary sinus was found in 25 patients,and draining chamber was right atrium in 13 cases, right ventricle in 12 cases. RSVA from noncoronary sinus was diagnosed in 8 patients,and the draining chamber was right atrium. Aortography defined the narrowest diameter at the rupture site was (6.4±1.7)mm. The ratio of Qp/Qs was 2.2±0.5,and the mean pressure of pulmonary artery was 24.0(21.2,33.7)mmHg(1 mmHg=0.133 kPa). One patient developed serious occluder related aortic regurgitation and underwent surgery, transcatheter closure was successfully performed in 32 patients. The success rate of transcatheter closure was 97.0%. Two types of device were used in the study including small-waist double-disk ventricular septal defect(VSD) occluders in 20 cases and patent ductus arteriosus(PDA) occluders in 12 cases. During a median follow-up of 73.5(28.3,89.5) months, there were no infective endocarditis, residual shunt, thrombosis, device displacement,serious aortic regurgitation, serious arrhythmia or death.At the last follow-up, the left atrial diameter((37.4±6.5) mm vs. (41.5±5.3)mm,P<0.01),right atrial diameter((42.4±3.0) mm vs. (48.5±6.0)mm,P<0.01), right ventricular diameter((22.2±3.8) mm vs. (27.7±7.2)mm,P<0.01) and left ventricular end-diastolic diameter((51.3±4.9) mm vs.(55.0±4.3)mm,P<0.01)measured by echocardiography were all smaller than pre-procedural level. Conclusion Transcatheter closure of RVSA is a safe and effective strategy and associated with a good long-term outcome.

Objective To evaluate the clinical safety, efficacy and long-term outcome of transcatheter occlusion for ruptured aortic sinus of valsalva aneurysm (RASA) into the right atrium.Methods Between January 2006 and April 2013, fifteen patients [11 males and 4 females,aged from 21 to 48 years with an mean age of (35.50±8.79) years] with RASA ruptured into the right atrium were enrolled in this study.Domestic made patent ductus arteriosus (applied in six patients) or small waist double-disk ventricular septal defect (applied in nine patients) occluders were used for transcatheter closure.All the patients were followed up for any change in cardiac rhythm,and residual shunt,occluders morphology and possible valve regurgitation by echocardiography.Results All RASA were confirmed by aortography,including eleven cases with rupture of right coronary sinus of valsalva and four cases with rupture of the noncoronary sinus of valsalva shunting into the right atrium.NYHA function class was(2.56±0.63)before the occlusion.Cardiac catheterization showed mean pulmonary arterial pressure and Qp/Qs ratio were (25.38±8.21)mmHg (1 mmHg=0.133 kPa) and 1.34-2.81(1.93±0.39), respectively.Aortic angiography showed that the RSA was 4-10(6.42±1.92)mm at its narrowest end.There was no serious complication during the operation and all the patients had successful transcatheter closure without residual shunt.After transcatheter RASA occlusion, mean pulmonary artery pressure decreased to (16.1±5.3) mmHg (P<0.05).The diameter of right atrium,right ventricle, left atrium and pulmonary artery diameter and left ventricular end-diastolic dimension all showed significant decrease (P<0.01).All patients were followed up for 35-132(78.6±28.57)months.All patients presented with a NYHA function class Ⅰ to Ⅱ cardiac function in their last follow up which was significantly improved compare to pre-occlusion level (P<0.01).There were no infective endocarditis,device displacement and embolism,serious aortic regurgitation,myocardial ischemia,serious arrhythmia or death in any of the patients during follow up.Conclusions Transcatheter closure of Valsalva aneurysm ruptured into right atrium with the domestic made patient ductus arteriosus and small-waist ventricular septal defect occluder is safe and effective with a good long term prognosis.

Objective To investigate the risk factors and early diagnosis of the severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus ( PDA ) . Methods Between February, 2011 and May, 2015, 80 patients with patent ductus arteriosus underwent percutaneous intervention occlusion were studied. Results Average age were ( 17. 5 ± 17. 1 ) years, 63 were females (78. 8%), mean weight were (35. 6 ± 20. 2)kg (from 6. 0 to 75. 0 kg), mean body surface area (BSA) were (1. 09 ±0. 44) m2(from 0. 32 to 1. 91 m2). A bolus of heparin calcium (80 U/kg) was administered by intravenous injection. The mean diameters of patent ductus arteriosus were 4 mm (from 2 to 18 mm), and the mean diameters of occluders were 12 mm (from 6 to 30 mm). 14 patients were found to have severe thrombocytopenia (PLT count﹤100 × 109/L). The reduction rate of platelet in 12 of 14 patients was more than 19%. The diameters of all occluders were equal to or more than 14 mm, the mean diameters of patent ductus arteriosus were 10 mm ( from 6 to 18 mm) and the mean diameters of occluders were 18 mm ( from 14 to 30 mm). All the 14 patients started to present progressive decrease in PLT count since the second day post procedure. Taking the selected occluder diameter greater than 14 mm as cut-off points in diagnosis of severe thrombocytopenia, the sensitivity was 100%, specificity was 68%, the positive predictive value was 40%, and the negative predictive value was 100%. Combined with the postprocedural second day complete blood count analysis and the platelet count decreased by 10% as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 93%, specificity was 67%, the positive predictive value was 65%, the negative predictive value was 93% . If taking the platelet count decreased by 7% on second day as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 100%, specificity was 57%, the positive predictive value was 61%, the negative predictive value was 100% . Logistic regression analysis discovered that risk factors of severe thrombocytopenia after PDA are procedural platelet count and occluder diameter. Conclusions The risk factors of severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus were the procedural reduction of platelet count and big occluder diameter. Patients with PDA who were inplanted with occluders equal to or bigger than 14 mm should retest the numbers of platelet on the second day after procedure and retest on third day if the numbers reduce on the second day, which may help in the prediction of severe thrombocytopenia.

Objective To investigate the therapeutic effect of severe thrombocytopenia in patients underwent transcatheter patent ductus arteriosus occlusion.Methods Clinical data of 80 pure patent ductus arteriosus patients who underwent interventional occlusion between February 2011 and November 2014 in General Hospital of Shenyang Military Region were retrospective analyzed.A bolus of heparin calcium (80 U/kg) was administered by intravenous injection during the procedure.Blood test was conducted in all patients before and after the procedure.The patients with both platelet reduction rate ≥ 5％ and platelet count ＜ 100 × 109/L(severe thrombocytopenia) were treated by following strategies:bed rest,avoidance of activities,intensive control of blood pressure through sodium nitroprusside administration,and inhibition of presumed immunological reaction by glucocorticoids and human immunoglobulin administration.Follow-up ended in May 2015.Results (1) A total of 54 cases (67.5％) were diagnosed as thrombocytopenia in second days after the procedure.The reduction rate of platelet in 41 out of 54 patients was equal or more than 5％ on the second day,and the diameter of occludes were equal or more than 14 mm in 35 out of these 54 patients.Severe thrombocytopenia occurred in 14 patients and the occluder diameter was ≥ 14 mm in these 14 patients.Severe thrombocytopenia did not occur in patients with occlude diameter ＜ 14 mm or with reduction rate of platelet ＜5％.(2) A total of 14 severe thrombocytopenia patients were treated.No bleeding events occurred in hospital and during the whole follow-up period.The the lowest value of platelet count of patients was seen on the 4.5 (3.8,6.0) days post the procedure.The lowest numbers were 16.5 (7.5,37.3) × 109/L.The platelet count of patients restored to more than 30 × 109/L on the 8.0 (4.8,9.5) days and restore to more than 100 × 109/L on the 12.0 (9.8,39.3) days post procedure.The average hospitalization day of the patients were 15.5 (11.8,21.5) days.The platelet counts of severe thrombocytopenia patients were normal during the end of follow up (median 3.8 (1.0,4.9)years).Conclusion The patients with very severe thrombocytopenia after transcatheter occlusion of patent ductus arteriosus are associated with a benign outcome in case of timely and proper treatment.

Photosynthetic and growth characteristics of Angelica dahurica were studied in order to clarity the relations of photosynthesis, growth and root dry weight, and provide a theoretical basis for its cultivation. Photosynthesis and growth indexes were meas- ured every 25 days. The contents of chlorophyll a, b, a + b, soluble protein and the activities of Hill reaction, Ca(2+)-ATPase, Mg(2+)-ATPase had an increasing trend; They had the highest value in leaf high-speed growth period. Then, they were decreased in root high- speed growth period. The root dry weight showed negative corelation with photosynthetic characteristics indexes except stomatal con- ductance, however, the negative corelation only from net photosynthetic rate and Ca(2+)-ATPase were significant. The vegetative growth period of spring sowing A. dahuricia was divided into three phases: seedling period, leaf high-speed growth period and root high-speed growth period. The root dry weight showed a significantly positive corelation with the root diameter, leaf dry weight, shoot dry weight, aboveground dry weight. There was the competitive relation between aboveground and underground, so underground growth could be es- timated from leaf area and shoot dimeter.
Adenosine Triphosphatases ; metabolism ; Angelica ; enzymology ; growth & development ; metabolism ; Chlorophyll ; metabolism ; Photosynthesis ; Plant Proteins ; metabolism ; Plant Roots ; growth & development ; Seasons

Objective to investigate the relationship between the size of ostium secondary atrial septal defect (ASD) and the pulmonary arterial pressure (PAP) in children less than 5 years of age. Methods During the period from April 2000 to January 2011, a total of 189 child patients less than 5 years of age with ostium secondary ASD were admitted to General Hospital of Shenyang Military Command. Under general anaesthesia with ketamine cardiac catheterization was performed, PAP was measured, and percutaneous occlusion of ASD was carried out in all patients. The clinical indexes, including sex, age, body height, body weight, body surface area, diameter of ASD defect, heart- to- thorax ratio, the systolic, diastolic and mean pressure of the pulmonary artery, etc. were determined. The patients were followed up for one year and postoperative cardiac ultrasonography was performed to check the results. The patients were divided into groups according to the defect size. Results The 189 patients consisted of 77 males and 112 females with a male-to-female ratio of 1 ∶ 1.5. The mean age was (4.1 ± 0.9) years old, ranging from 2 to 5 years old. The mean weight was (17.2 ± 3.6) kg, ranging from 10.0 to 30.0 kg. The mean height was (104.9 ± 9.2) cm, ranging from 77 to 135 cm. The mean body surface area (BSA) was (0.71 ± 0.10) m2, ranging from 0.46 to 1.02 m2. The mean size of ASD was (12.6 ± 4.8) mm, ranging from 5 to 29 mm. The mean size of ASD, which was modified by BSA, was (18.0 ± 7.0) mm/m2, ranging from 5.3 to 38.9 mm/m2. The mean systolic PAP was (41.1 ± 8.9) mmHg with a range of 15 - 67 mmHg. The mean diastolic PAP was (16.8 ± 6.5) mmHg with a range of 3 - 45 mmHg. The mean PAP was (24.9 ± 6.7) mmHg with a range of 12 - 48 mmHg. One hundred and fifty- nine patients (89.4%) had pulmonary arterial hypertension (PAH) which was determined by right heart catheterization, but no patient showed PAH when the pulmonary arterial pressure was measured by echocardiography before the procedure as well as 1, 3, 6, 12 months after the procedure. No definite correlation existed between the size of ASD and the pulmonary artery pressure (P > 0.05). Conclusion Pulmonary artery pressure measured by right heart catheterization has no definite correlation with the size of ASD in children less than 5 years of age. Pulmonary artery pressure obtained from right heart catheterization is higher than that determined by cardiac ultrasonography, which may be caused by the effect of ketamine when general anaesthesia is used in performing right heart catheterization.