Cloning and Expression Patterns of Coumarate-3-hydroxylase Gene from Angelica sinensis and Its Correlation with Content of Ferulic Acid / 中国实验方剂学杂志

ObjectiveTo clone coumarate-3-hydroxylase gene （C3H） from Angelica sinensis， and analyze the correlation between its bioinformatics， expression patterns and content of ferulic acid， and to explore the functions of ASC3H. MethodReal-time polymerase chain reaction （Real-time PCR） was used to clone the full-length cDNA of ASC3H based on the transcriptome dataset of A. sinensis， and the bioinformatics analysis of the gene sequence was carried out. Real-time PCR and high performance liquid chromatography （HPLC） were used to determine relative expression of ASC3H and content of ferulic acid in different root tissues of A. sinensis （periderm， cortex and stele）. ResultThe open reading frame （ORF） of ASC3H （GenBank accession number： MN2550298） was 1 530 bp， encoding 509 amino acids， with a theoretical molecular weight of 57.86 kDa and an isoelectric point of 8.36. It was a hydrophilic protein that was located in the chloroplast with multiple phosphorylation sites and a transmembrane region， and contained a conserved domain CGYDWPKGYGPIINVW_P450 （383-399 aa） in cytochrome P450. Multiple amino acid sequence alignment analysis showed that ASC3H had high similarity with C3H from other plants， especially Ammi majus in Umbelliferae. The Real-time PCR revealed that ASC3H had different expressions in periderm， cortex and stele tissues of A. sinensis roots. It was found from HPLC that the cortex tissues had the highest content of ferulic acid， and the stele tissues had the lowest. ConclusionASC3H was successfully cloned from A. sinensis， and its sequence characteristics were understood more clearly， suggesting that ASC3H might be involved in the ferulic acid biosynthesis pathway of A. sinensis. This paper provided a basis for further studying the functions of the gene and exploring the biosynthesis and regulation mechanism of ferulic acid in A. sinensis， while laying the foundation for the genetic improvement of A. sinensis．

A circRNA-miRNA-mRNA network analysis underlying pathogenesis of human heart failure / 老年心脏病学杂志（英文版）

BACKGROUND@#The molecular mechanisms of heart failure (HF) are still poorly understood. Circular RNA (circRNA) has been discovered in the heart in increasing numbers of studies. The goal of this research is to learn more about the potential roles of circRNAs in HF.@*METHODS & RESULTS@#We used RNA sequencing data to identify the characteristics of circRNAs expressed in the heart and discovered that the majority of circRNAs screened were less than 2000 nt. Additionally, chromosomes One and Y had the most and least number of circRNAs, respectively. After excluding duplicate host genes and intergenic circRNAs, a total of 238 differentially expressed circRNAs (DECs) and 203 host genes were discovered. However, only four of the 203 host genes of DECs were examined in HF differentially expressed genes. Another study used Gene Oncology analysis of DECs host genes to elucidate the underlying pathogenesis of HF, and it found that binding and catalytic activity accounted for a large portion of DECs. Immune system, metabolism, and signal transduction pathways were significantly enriched. Furthermore, 1052 potentially regulated miRNAs from the top 40 DECs were collected to build a circRNA-miRNA network, and it was discovered that 470 miRNAs can be regulated by multiple circRNAs, while others are regulated by a single circRNA. In addition, a comparison of the top 10 mRNAs in HF and their targeted miRNAs revealed that DDX3Y and UTY were regulated by the most and least circRNA, respectively.@*CONCLUSION@#These findings demonstrated circRNAs have species and tissue specific expression patterns; while circRNA expression is independent on host genes, the same types of genes in DECs and DEGs worked in HF. Our findings would contribute to a better understanding of the critical roles of circRNAs and lay the groundwork for future studies of HF molecular functions.

Prognostic value of the number and anatomical distribution of tumor deposits in patients with gastric cancer without lymph node metastasis / 中华普通外科杂志

Objective:To explore the prognostic value of tumor deposits (TD) by number and anatomical distribution in gastric cancer (GC) patients without lymph node metastasis.Methods:From Aug 2012 to Aug 2018 all 91 GC patients undergoing radical gastrectomy and without nodal metastasis at Yijishan Hospital of Wannan Medical College were enrolled in this study. Patients were divided into L1, L2, and L3 groups according to the number of TD and into Q1 and Q2 groups according to the anatomical regions of the TD.Results:The 3-year overall survival (OS) rates of groups L1, L2, and L3 were 58.9%, 52.1%, and 31.5%, respectively ( χ2=9.769, P=0.008). The 3-year OS rates of groups Q1 and Q2 were 58.9% and 7.1% ( χ2=46.310, P<0.001). The number of TD, their distribution, neural invasion, vascular invasion, tumor size, and pT stage were all related to prognosis by univariate analysis (all P<0.05). Tumor size>4 cm ( HR=2.460, 95% CI:1.307-4.629, P=0.005), distribution of TD (non-perigastric)( HR=3.959, 95% CI:2.077-7.545, P<0.001), neural invasion ( HR=4.299,95% CI:1.953-9.461, P<0.001), and pT 4 stage ( HR=2.283, 95% CI:1.250-4.171, P=0.007) were independent risk factors for prognosis by multivariate analysis. Conclusion:The distribution of TD (non-perigastric) is an independent risk factor for poor prognosis in gastric cancer patients after radical gastrectomy and with negative lymph node metastasis.

Application of the fetal adrenal volume evaluated by three-dimensional ultrasound in fetuses with intrauterine growth restriction and its correlation with adverse perinatal outcomes / 中华超声影像学杂志

Objective:To evaluate the fetal adrenal gland volume (AGV) and corrected adrenal gland volume (cAGV) in intrauterine growth restriction (IUGR) fetuses and observe their associations with the adverse perinatal outcomes.Methods:From February 2021 to August 2022, 32 IUGR fetuses who underwent fetal ultrasound examination in the Second Xiangya Hospital of Central South University were prospectively selected as the IUGR group, and 32 normal fetuses matched for gestational age during the same period were selected as the control group. Three-dimensional ultrasound was used to obtain fetal adrenal volume images, and the virtual organ computer-aided analysis (VOCAL) was used to measure AGV, then the cAGV was calculated. The values of AGV and cAGV were appropriately compared between the IUGR and the control groups. The pregnancy outcomes were noted. Multiple logistic regression analysis was employed to evaluate the relationship between the cAGV and adverse perinatal outcomes in IUGR fetus, with maternal age and the CPR included as covariates to control for confounding factors.Results:A total of 32 fetuses with IUGR and 32 controls were involved in this prospective study. There was no significant difference in the AGV between these two groups ( P=0.417). The cAGV of the IUGR fetus was substantially larger than that of the normal fetus ( P=0.034). In the multivariate logistic regression analysis, after adjusting for maternal age and fetal CPR, the fetal cAGV was noticeably associated with the fetal distress (adjusted OR=0.005, 95% CI=0.000-0.587, P=0.029) and the total adverse perinatal outcomes (adjusted OR=0.014, 95% CI=0.000-0.475, P=0.018). Conclusions:The value of cAGV is increased in the IUGR fetuses and associated with adverse perinatal outcomes. The evaluation of fetal AGV could be beneficial to monitoring and managing IUGR fetuses.

Predictive value of early lactate/albumin ratio in the prognosis of sepsis / 中华危重病急救医学

Objective:To investigate the prognostic value of early serum lactate, albumin, and lactate/albumin ratio (L/A) on the 28-day prognosis of adult patients with sepsis.Methods:A retrospective cohort study was conducted among adult patients with sepsis admitted to the First Affiliated Hospital of Xinjiang Medical University from January to December in 2020. Gender, age, comorbidities, lactate within 24 hours of admission, albumin, L/A, interleukin-6 (IL-6), procalcitonin (PCT), C-reactive protein (CRP) and 28-day prognosis were recorded. The receiver operator characteristic curve (ROC curve) was drawn to analyze the predictive value of lactate, albumin and L/A for 28-day mortality in patients with sepsis. Subgroup analysis of patients was performed according to the best cut-off value, Kaplan-Meier survival curves were drawn, and the 28-day cumulative survival of patients with sepsis was analyzed.Results:A total of 274 patients with sepsis were included, and 122 patients died at 28 days, with a 28-day mortality of 44.53%. Compared with the survival group, the age, the proportion of pulmonary infection, the proportion of shock, lactate, L/A and IL-6 in the death group were significantly increased, and albumin was significantly decreased [age (years): 65 (51, 79) vs. 57 (48, 73), pulmonary infection: 75.4% vs. 53.3%, shock: 37.7% vs. 15.1%, lactate (mmol/L): 4.76 (2.95, 9.23) vs. 2.21 (1.44, 3.19), L/A: 0.18 (0.10, 0.35) vs. 0.08 (0.05, 0.11), IL-6 (ng/L): 337.00 (97.73, 2 318.50) vs. 55.88 (25.26, 150.65), albumin (g/L): 27.68 (21.02, 33.03) vs. 29.62 (25.25, 34.23), all P < 0.05]. The area under the ROC curve (AUC) and 95% confidence interval (95% CI) of lactate, albumin, and L/A were 0.794 (95% CI was 0.741-0.840), 0.589 (95% CI was 0.528-0.647), 0.807 (95% CI was 0.755-0.852) for predicting 28-day mortality in sepsis patients. The optimal diagnostic cut-off value of lactate was 4.07 mmol/L, the sensitivity was 57.38%, the specificity was 92.76%. The optimal diagnostic cut-off value of albumin was 22.28 g/L, the sensitivity was 31.15%, the specificity was 92.76%. The optimal diagnostic cut-off of L/A was 0.16, the sensitivity was 54.92%, and the specificity was 95.39%. Subgroup analysis showed that the 28-day mortality of sepsis patients in the L/A > 0.16 group was significantly higher than that in the L/A ≤ 0.16 group [90.5% (67/74) vs. 27.5% (55/200), P < 0.001]. The 28-day mortality of sepsis patients in the albumin ≤ 22.28 g/L group was significantly higher than that in the albumin > 22.28 g/L group [77.6% (38/49) vs. 37.3% (84/225), P < 0.001]. The 28-day mortality in the group with lactate > 4.07 mmol/L was significantly higher than that in the group with lactate ≤ 4.07 mmol/L [86.4% (70/81) vs. 26.9% (52/193), P < 0.001]. The three were consistent with the analysis results of Kaplan-Meier survival curve. Conclusion:The early serum lactate, albumin, and L/A were all valuable in predicting the 28-day prognosis of patients with sepsis, and L/A was better than lactate and albumin.

Comparison of short-term outcomes between full-port robotic and thoracoscopic mediastinal tumor resection: A propensity score matching study / 中国胸心血管外科临床杂志

@#Objective    To analyze and compare the perioperative efficacy difference between full-port Da Vinci robotic surgery and thoracoscopic surgery in patients with mediastinal tumor resection. Methods    The data of 232 patients with mediastinal tumors treated by the same operator in the Department of Thoracic Surgery of the Second Affiliated Hospital of Harbin Medical University were included. There were 103 (44.4%) males and 129 (55.6%) females, with an average age of 49.7 years. According to the surgical methods, they were divided into a robot-assisted thoracic surgery (RATS) group (n=113) and a video-assisted thoracoscopic surgery (VATS) group (n=119). After 1 : 1 propensity score matching, 57 patients in the RATS group and 57 patients in the VATS group were obtained. Results    The RATS group was better than the VATS group in the visual analogue scale pain score on the first day after the surgery [3.0 (2.0, 4.0) points vs. 4.0 (3.0, 5.0) points], postoperative hospital stay time [4.0 (3.0, 5.5) d vs. 6.0 (5.0, 7.0) d] and postoperative catheterization time [2.0 (2.0, 3.0) d vs. 3.0 (3.0, 4.0) d] (all P<0.05). There was no statistical difference between the two groups in terms of intraoperative blood loss, postoperative complications, postoperative thoracic closed drainage catheter placement rate or postoperative total drainage volume (all P>0.05). The total hospitalization costs [51 271.0 (44 166.0, 57 152.0) yuan vs. 35 814.0 (33 418.0, 39 312.0) yuan], operation costs [37 659.0 (32 217.0, 41 511.0) yuan vs. 19 640.0 (17 008.0, 21 421.0) yuan], anesthesia costs [3 307.0 (2 530.0, 3 823.0) yuan vs. 2 059.0 (1 577.0, 2 887.0) yuan] and drug and examination costs [9 241.0 (7 987.0, 12 332.0) yuan vs. 14 143.0 (11 620.0, 16 750.0) yuan] in the RATS group was higher than those in the VATS group (all P<0.05). Conclusion    Robotic surgery and thoracoscopic surgery can be done safely and effectively. Compared with thoracoscopic surgery, robotic surgery has less postoperative pain, shorter tube-carrying time, and less postoperative hospital stay, which can significantly speed up the postoperative recovery of patients. However, the cost of robotic surgery is higher than that of thoracoscopic surgery, which increases the economic burden of patients and is also one of the main reasons for preventing the popularization of robotic surgery.

Identification of Andrographis Herba and its common products using mini-barcode / 中国天然药物

Andrographis Herba, the aerial part of Andrographis paniculata (Burm. f.) Wall. ex Nees (Acanthaceae), has a wide geographic distribution and has been used for the treatment of fever, cold, inflammation, and other infectious diseases. In markets, sellers and buyers commonly inadvertently confuse with related species. In addition, most Chinese medicinal herbs are subjected to traditional processing procedures, such as steaming and boiling, before they are sold at dispensaries; therefore, it is very difficult to identify Andrographis Herba when it is processed into Chinese medicines. The identification of species and processed medicinal materials is a growing issue in the marketplace. However, conventional methods of identification have limitations, while DNA barcoding has received considerable attention as a new potential means to identify species and processed medicinal materials. In this study, 17 standard reference materials of A. paniculata, 2 standard decoctions, 27 commercial products and two adulterants were collected. Based on the ITS2 sequence, it could successfully identify A. paniculata and adulterants. Moreover, a nucleotide signature consisting of 71 bp was designed, this sequence is highly conserved and specific within A. paniculata while divergent among other species. Then, we used these new primers to amplify the nucleotide signature region from processed materials. In conclusion, the DNA barcoding method developed in the present study for authenticating A. paniculata is rapid and cost-effective. It can be used in the future to guarantee the quality of Andrographis Herba of each regulatory link for clinical use.

Comparison of initial periodontal therapy and its correlation with white blood cell level in periodontitis patients with or without diabetes mellitus / 北京大学学报(医学版)

OBJECTIVE@#To compare the clinical efficacy of initial periodontal therapy in periodontitis patients with or without type 2 diabetes mellitus and its correlation with white blood cell counts.@*METHODS@#In this study, 32 chronic periodontitis patients without systemic disease (CP group) and 27 chronic periodontitis patients with type 2 diabetes mellitus (CP+DM group) were enrolled. At admission, all the patients went through periodontal examination and fasting blood examination(baseline). Probing depth (PD), attachment loss (AL), bleeding index (BI), plaque index (PLI), white blood cells (WBC) counts and fasting blood glucose (FBG) were recorded respectively, while hemoglobin A1c (HbA1c) was recorded only in CP+DM group. After that, initial periodontal therapy was performed. All the tests were repeated 3 and 6 months after treatment. The changes of periodontal clinical indexes and WBC levels were compared between the two groups before and after treatment, and the correlation between WBC and periodontal clinical indexes and glucose metabolism indexes were analyzed by generalized linear mixed model.@*RESULTS@#At baseline, the periodontal inflammation and destruction were similar in CP and CP+DM group, but the WBC level was significantly higher in CP+DM groups [(6.01±1.26)×109/L vs. (7.14±1.99)×109/L, P=0.01]. After 3 and 6 months of initial periodontal therapy, the mean PD, AL, BI, and PLI in CP+DM and CP groups were significantly lower than the baseline, and the PD in CP+DM group was further decreased by 6 months compared with 3 months [(3.33±0.62) mm vs. (3.61±0.60) mm, P < 0.05]. However, none of these periodontal indexes showed significant difference between the two groups by 3 or 6 months. In CP+DM group, HbA1c at 3 months and 6 months were significantly lower than the baseline [(7.09±0.79)% vs. (7.64±1.16)%, P < 0.05; (7.06±0.78)% vs. (7.64±1.16)%, P < 0.05], and FBG was significantly lower than the baseline by 6 months [(7.35±1.14) mmol/L vs. (8.40±1.43) mmol/L, P < 0.05]. The WBC level in CP group was significantly lower than the baseline level by 3 months [(5.35±1.37)×109/L vs. (6.01±1.26)×109/L, P < 0.05], while that in CP+DM group was significantly lower than the baseline level by 6 months [(6.00±1.37)×109/L vs. (7.14±1.99)×109/L, P < 0.05]. The analysis of genera-lized linear mixed model showed that WBC level was significantly positively correlated with PD and FBG (P < 0.05).@*CONCLUSION@#Initial periodontal therapy can effectively improve the periodontal clinical status of patients with or without type 2 diabetes mellitus, and have benefits on glycemic control in diabetic patients. However, the response of periodontal indexes and WBC level to initial therapy were relatively delayed in diabetic patients. WBC plays an important role in the correlation between diabetes mellitus and periodontitis.

The effect of large-conductance calcium-activated potassium channels on the migration of pericytes in the mice of senile cochlear stria vascularis / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate whether large conductance calcium-activated potassium channel (BK(Ca)) was involved in the migration of pericytes (PC) in the mice of senile cochlear stria vascularis capillaries PC. Methods: C57BL/6J mice were divided into 3-month (n=10) and 12-month groups (n=10). Auditory brainstem response (ABR) was used to test the hearing threshold of each group. The immunofluorescence was used to detect the expression changes of osteopontin (OPN) and β-BK(Ca) channels on cochlear stria vascularis PC. The morphological changes of perivascular cells in cochlea were observed by transmission electron microscope (TEM). Cell experiment: The PC, which were in the stria vascularis of the cochlea were primary cultured and identified. A cell senile model was made with D-gal. The appropriate intervention concentration of low galactose (D-gal) was determined by CCK8. β-galactosidase (SA-β-gal) staining was used to evaluate the cell decrept level. The change of BK(Ca) channels current on PC were recorded by whole cell patch clamp technique. The expression of BK(Ca) channels on PC was detected by immunofluorescence. The migration and invasion ability of two groups were detected by using Scratch test and Transwell. The levels of OPN and β-BK(Ca) channels were detected by Western blot. SPSS 22.0 software was used to analyze the data. Results: The ABR threshold in the 12-month group was higher than 3-month group (t=12.66, P<0.01). In the 12-month group, the expression of β-BK(Ca) channel was lower and the expression of OPN was increased (t=14.64, P<0.01; t=20.73, P<0.01). In TEM, cochlear stria vascularis PC were tightly connected to endothelial cells in 3-month group, while PC were loosely connected to endothelial cells or PC soma were separated from the capillary in 12-month group. Cell experiment: The positive rate of PC in the primary cultured cochlear stria vascularis is above 95%. Compared with the SA-β-gal stained cells in the control group, the positive rate of 15 mg/ml D-gal intervention PC was 85% (t=36.90, P<0.01). Whole cell patch clamp BK(Ca) channels current decreased in the D-gal group compared with the young group PC (t=12.18, P<0.05). The OPN expression in the senile group was higher than control group (t=16.30, P<0.01), while the β-BK(Ca) channels expression was decreased (t=11.98, P<0.01; t=15.72, P<0.05), and migration ability raised (t=7.91, P<0.01;t=7.59, P<0.01). After intervened of BK(Ca) channels specific blocker IBTX in the D-gal group, the expression of OPN and migration were increased (t=4.26, P<0.05; t=5.88, P<0.01; t=21.97, P<0.01). Conclusion: PC migration capacity were increased during the senile period, and the expression of β-BK(Ca) channel was decreased. The administration of IBTX, a specific blocker of BK(Ca) channel, at the cell level could increase the migration capacity, suggesting that BK(Ca) might be involved in the migration of PC in the stria vascularis of the aging cochlea.

Effect of microvascular pericytes of cochlear stria vascularis on endothelial cell permeability in C57BL/6J mice / 中华耳鼻咽喉头颈外科杂志

Objective: To study the changes in the permeability of the blood labyrinth barrier of the aging cochlea in mice, and to establish a non-contact co-culture model of endothelial cells (EC) and pericytes (PC) to furtherly investigate the cochlear stria vascularis microvascular pericytes impact on the permeability of endothelial cells. Methods: C57BL/6J mice were divided into two groups, three months old as young group, 12 months old as senile group. Cell experiment was divided into four groups, EC group, EC+PC co-culture group, D-gal+EC group and D-gal+EC+PC co-culture group. Auditory brainstem response (auditory brain response, ABR) was used to detect the auditory function of the two groups of mice. Evans blue staining was applied to detect the permeability of the cochlear blood labyrinth barrier of the two groups of mice. Transmission electron microscopy was used to observe the ultrastructure of blood labyrinth barrier endothelial cells, pericytes and tight junctions in the two groups of mice. Immunohistochemistry was used to detect the expression levels of tight junction proteins in the stria vascularis of the cochlea of the two groups of mice. Transwell chamber was used to detect the permeability of endothelial cells. Western blot and immunofluorescence technology were used to detect the expression level of tight junction protein on endothelial cells. SPSS 20.0 software was used to analyze the data. Results: Compared with the young group, the ABR threshold of the aging group was significantly increased, the latency of wave I was prolonged (t=10.25, P<0.01;t=5.61, P<0.05), the permeability of the cochlear blood labyrinth barrier was increased and the expression of tight junction protein on the vascular stria was decreased (P<0.05). The cochlear ultrastructure showed that the cochlear vascular stria microvascular lumen was deformed, the basement membrane thickened and the tight junction gap between endothelium enlarged. The positive rate of ECs and PCs in primary culture was more than 95%. The cells induced by 15 g/L D-gal were determined to be senescent cells. Compared with EC group, the expression of tight junction protein in endothelial cells of D-gal+EC group decreased（t=7.42，P<0.01；t=13.19，P<0.05）and the permeability increased (t=11.17, P<0.01). In the co-culture group, the expression of tight junction protein between endothelial cells in EC+PC co-culture group and D-gal+EC+PC co-culture group increased and the permeability decreased. Conclusions: In aging mice, the permeability of cochlear blood labyrinth barrier will increase and the level of tight junction protein will decrease; in aging state, cochlear vascular stria microvascular pericytes may affect endothelial cell permeability by regulating the expression of tight junction protein.

Study on renal artery blood flow parameters of fetuses with isolated borderline oligohydramnios and maternal and infant pregnancy outcomes / 中华超声影像学杂志

Objective:To evaluate the changes of fetal renal artery blood flow parameters in fetuses with isolated borderline oligohydramnios (IBO) in the middle and third trimesters by Doppler ultrasound, and to assess its correlations with maternal and infant pregnancy outcomes.Methods:Twenty-seven IBO fetuses (IBO group) and 27 gestational age-matched normal fetuses (control group) from April to October 2019 in the Second Xiangya Hospital of Central South University underwent prenatal ultrasound examination during the middle and third trimesters. Renal artery blood flow parameters, including renal artery pulsatility index (RAPI), volume corrected renal artery pulsatility index (vcRAPI) and pregnancy outcomes were measured and compared between the two groups. Once diagnosed IBO, patients were recommended to the obstetric clinic for consultation and intervention. The correlation between RAPI, vcRAPI measured before intervention and prepartum amniotic fluid volume and pregnancy outcomes was analyzed, the ROC curve was plotted to find the better predictor.Results:The vcRAPI of the IBO group was higher than that of the control group ( P=0.015). In the IBO group, the vcRAPI measured before intervention was higer in those fetuses who were still IBO before delivery( P=0.048). In the IBO group, the correlation of the vcRAPI measured before intervention and IBO before delivery was statistically significant ( OR=2.41, 95% CI=1.06-5.43, P=0.035). The ROC curve showed that the sensitivity of vcRAPI to IBO was 0.67, the specificity was 0.75( P=0.002). Conclusions:Compared with RAPI, The vcRAPI may reflect the increase in fetal renal artery perfusion resistance of IBO group more timely. The higher vcRAPI before intervention in the IBO group have difficulty in recovering amniotic fluid volume before delivery.Increased vcRAPI is a better predictor of IBO before delivery.

Detection of measles virus infection and proliferation by real-time quantitative PCR / 上海预防医学

Objective A SYBR-Green real-time quantitative PCR(RT-qPCR)method was set up to detect the infection and proliferation of measles virus, which could be useful in virus titer determination. Methods In this project, we used a 405 bp fragment of the N gene of measles virus as a target sequence and constructed a plasmid to establish the standard curve in absolute quantitative experiment. We then used this method to obtain the proliferation curve of measles virus and to detect the virus proliferation at different MOI. Results There was a linear relationship between the virus copy number and the titer of the measles virus reference at the range of 6 to 2 lgCCID50/mL, with a correlation coefficient (r) of 0.991(P < 0.01). Based on the analysis of virus proliferation curve, measles virus mainly proliferated intracellularly within 48 h after its entering the cell. There was no detected increase in viral RNA level in the first 24 h, suggesting the virus was in a silent period in the cell. After 24 h, the virus expanded in large numbers and entered the exponential growth phase. The intracellular viral RNA level reached the plateau phase after its peak at 96 h. The virus secreted to the outside of the cell entered the exponential growth phase starting from 48 h, peaked at 144 h, then followed by plateau phase. Conclusion A SYBR-Green RT-qPCR method is established and used to monitor virus proliferation. Our result is helpful in understanding of the proliferation and secretion of measles virus in cells and provides experimental basis for detection of live attenuated virus titers.

Inhibitory Study of Huayu Jiedu Prescription Medicated Serum on Invasion and Migration of Human Lung Cancer Cell Line H1299 by JAK2/STAT3 Pathway / 中国实验方剂学杂志

Objective:To investigate the effect of Huayu Jiedu prescription medicated serum（HJRMS）on the proliferation， invasion and migration of human lung cancer cells （H1299 cells） and its mechanism. Method:Cell counting kit-8 （CCK-8） method was used to detect the inhibitory effect of HJRMS on the proliferation of lung cancer cells， the effect of HJRMS on the invasion and migration of H1299 cells were determined by Transwell assay and wound healing assay. The protein expressions of Janus kinase 2 （JAK2）， signal transduction and activation transcription factor 3 （STAT3）， phosphorylated JAK2（p-JAK2） and phosphorylated STAT3 （p-STAT3） were detected by Western blot， the mRNA expression levels of JAK2 and STAT3 were detected by Real-time quantitative polymerase chain reaction（Real-time PCR）. Result:① Compared with control group， the proliferation of H1299 cells was significantly inhibited after treatment with 1%~16%HJRMS serum for 24， 48 h， respectively（<italic>P</italic><0.01）， and showed a certain concentration dependence. ② After treatment with HJRMS for 24 h， the scratch healing ability of cells in the 4%，8%HJRMS serum groups was inhibited（<italic>P</italic><0.05，<italic>P</italic><0.01）. ③ Compared with control group， the membrane permeability of H1299 cells in invasion and migration experiments in 2%，4%，8%HJRMS serum groups was decreased significantly（<italic>P</italic><0.05，<italic>P</italic><0.01）. ④ Western blot showed that compared with control group， 4%，8%HJRMS serum groups inhibited the expression of JAK2/STAT3 signaling pathway related proteins （JAK2， p-JAK2， STAT3， and p-STAT3） in lung cancer H1299 cells（<italic>P</italic><0.05， <italic>P</italic><0.01）. ⑤ Compared with control group， the mRNA expression levels of JAK2 and STAT3 in lung cancer H1299 cells treated with 8%HJRMS for 24 h decreased significantly （<italic>P</italic><0.05）. Conclusion:The HJRMS can inhibit the proliferation， invasion and migration of lung cancer H1299 cells， and its mechanism may be related to the inhibition of JAK2/STAT3 signaling pathway.

Correlation between white matter hyperintensities and stroke etiology classification in patients with acute isolated penetrating artery territory infarction / 国际脑血管病杂志

Objective:To investigate the correlation between white matter hyperintensities (WMHs) and stroke etiology classification in patients with acute isolated penetrating artery territory infarction.Methods:Patients with first-ever acute isolated penetrating artery territory infarction admitted to the Department of Neurology, the Affiliated Hospital of Xuzhou Medical University from January 2017 to May 2020 were enrolled retrospectively. According to the Chinese Ischemic Stroke Subclassification (CISS) system, they were divided into large artery atherosclerosis (LAA) and perforating artery disease (PAD). According to the distribution of infarcts, they were divided into lenticulostriate artery (LSA) territory infarction and paramedian pontine artery (PPA) territory infarction. The demographics, vascular risk factors, baseline clinical data, WMHs location, and Fazekas Scale scores were documented. Multivariate logistic regression analysis was used to identify the independent influencing factors of stroke etiology classification. Results:A total of 440 patients with acute isolated penetrating artery territory infarction were enrolled, including 120 (27.3%) in the LAA group, and 320 (72.7%) in the PAD group; 213 (48.4%) with LSA territory infarction, and 227 (51.6%) with PPA territory infarction. The proportion of patients with total Fazekas score 3-6 and periventricular WMHs (PWMHs) score 2-3 in the PAD group was significantly higher than those in the LAA group (all P<0.05). In patients with LSA territory infarction, the proportion of the patients with hypertension, WMHs total Fazekas score 3-6 and PWMHs score 2-3 in PAD subgroup was significantly higher than those in the LAA subgroup, while the proportion of the patients with hyperlipidemia was significantly lower than that in LAA subgroup (all P<0.05). In patients with PPA territory infarction, the levels of low-density lipoprotein cholesterol and homocysteine in the PAD subgroup were significantly lower than those in the LAA subgroup. Multivariate logistic regression analysis showed that PWMHs score 2-3 was an independent correlation factor of PAD (odds ratio [ OR] 2.220, 95% confidence interval [ CI] 1.085-4.541; P=0.029). In patients with LSA territory infarction, hyperlipidemia was independently correlated with LAA ( OR 0.432, 95% CI 0.192-0.972; P=0.042), and PWMHs score 2-3 was independently correlated with PAD ( OR 3.846, 95% CI 1.193-12.397; P=0.024). In patients with PPA territory infarction, higher low-density lipoprotein cholesterol ( OR 0.660, 95% CI 0.494-0.883; P=0.005), homocysteine ( OR 0.958, 95% CI 0.930-0.987; P=0.005) and C-reactive protein ( OR 0.987, 95% CI 0.977-0.997; P=0.008) were independently correlated with LAA. Conclusions:WMHs are common in patients with acute isolated perforating territory infarction caused by LAA and PAD, and more severe PWMHs suggest that PAD is more likely to be the cause of the acute isolated perforating territory infarction, especially in patients with LSA territory infarction.

Z-scores analysis of kidney volume in normal fetuses in second and third trimester / 中华超声影像学杂志

Objective:To explore the variation of renal volume with gestational age(GA), biparietal diameter(BPD), abdominal circumference(AC), femur length (FL) and the Z value calculation formula.Methods:Five hundred and eighty singleton pregnancy women from 20th to 38th gestational weeks in Second Xiangya Hospital of Central South University were enrolled. Two dimensional ultrasound was performed to record fetal growth parameters and renal longitudinal, transverse, anteroposterior diameters. Renal volume was calculated using ellipsoid volume equation: Volume =6/π×length×width×thickness. Using GA, BPD, AC, FL as independent variables and renal volume as dependent variables, the regression analyses of the mean and the standard deviation(SD) for each parameter were calculated separately. The Z-scores were calculated by the formula: Z-scores=(actual measurements of renal volume-predictive value of renal volume)/SD of predictive value.Results:Five hundred and forty-three cases of normal singleton fetuses were involved.The left and right renal volume were increased along with GA, BPD, AC, FL. Quadratic polynomial regression equations were each fitted to the models( r=0.775, 0.771, 0.811, 0.738, P<0.001; r=0.747, 0.735, 0.754, 0.745, P<0.001, respectively). The SD increased with the increases of independent variables and could be modeled with a simple linear regression( r=0.146, 0.225, 0.071, 0.155, P<0.001; r=0.091, 0.157, 0.091, 0.123, P<0.001, respectively). Z-scores were calculated and proved normally distributed after Shapiro-Wilk test( P>0.05). Conclusions:The normal Z-score reference range of fetal renal volume is established to help assess the normal renal growth and identify fetal renal abnormalities.

Volume-corrected renal artery pulsatility index in normal fetuses during the second and third trimester of gestation / 中华超声影像学杂志

Objective:To measure the normal single fetal volume-corrected renal artery pulsatility index (vcRAPI) in the middle and late pregnancy, and to determine the normal reference range of vcRAPI and to observe its correlation with gestational age.Methods:The sagittal and transverse sections of the kidneys of normal single pregnant fetuses at 22 to 36 gestational weeks in the Second Xiangya Hospital from January to July 2019 were prospectively obtained by two-dimensional ultrasound. Estimated renal volume was calculated as length× transverse × antero-posterior×π/6. Pulse Doppler was used to detect the renal artery pulsatility index (RAPI) at the renal hilum in coronary section. The vcRAPI was obtained by dividing the RAPI by the ipsilateral estimated renal volume. The normal reference range of vcRAPI was established, and the scatter diagram of its relationship with gestational age was drawn. Multiple regression models were fitted to select the best fitting curve and to obtain the relevant formulas.Results:A total of 540 normal fetuses were enrolled and the normal reference value range of vcRAPI was established. The vcRAPI of both kidneys was negatively correlated with gestational age ( r=-0.793, -0.780; all P<0.001). The left kidney parameters and gestational age were best fitted with S-curve, and the right kidney parameters were best fitted with power curve. There was no difference between the right and left kidney vcRAPI values( P>0.05). Conclusions:The vcRAPI of normal fetuses decreases with gestational age, indicating that renal perfusion increases with gestational age. In this study, normal reference range of vcRAPI is established to provide reference basis for prediction and monitoring of complicated pregnancy.

Feasibility of echocardiography-guided repeated intraventricular blood sampling in mice / 中华心血管病杂志

Objective@#To investigate the feasibility of echocardiography-guided closed-chest repeated intraventricular blood sampling in mice, and to clarify the maximum blood volume that can be collected by this method, and whether the method can be used for long-term repeated blood collection in mice.@*Methods@#Twenty-four male C57BL/6J mice (10-14 weeks old) were divided into the terminal experiment group (n=4, for investigating the maximum blood amount that could be sampled at one time), the repeated 0.5 ml blood collection group (n=10, sampling 0.5 ml whole blood each time, once every two days for consecutive 4 weeks), and the repeated 0.75 ml blood collection group (n=10, sampling 0.75 ml whole blood each time, once every two days for consecutive 4 weeks). High-frequency echocardiography was used to display the largest section of the left ventricle, guiding the insulin syringe needle through the thorax into the left ventricle for blood collection. In the repeated 0.5 ml blood collection group, echocardiography was used to detect the cardiac structure and function before blood collection, three minutes after blood collection, and one week after the last (the 14th) blood collection.@*Results@#We successfully performed echocardiography-guided closed-chest intraventricular blood sampling, with an average operating time (88±19)s per mouse, and a maximum blood volume (1.43±0.11)ml per mouse. In the repeated 0.5 ml blood collection group, heart rate, left ventricular ejection fraction, left ventricular fractional shortening, left ventricular end-diastolic dimension and left ventricular posterior wall end-diastolic thickness remained uncganged before the first blood collection and after 4 weeks of repeated blood collection (all P>0.05). No death in the repeated 0.5 ml blood collection group. However, in the 0.75 ml blood collection group, two mice died before the end point.@*Conclusions@#The echocardiography-guided closed-chest intraventricular blood sampling is a safe, minimally invasive, convenient and efficient method, and can be used repeatedly for long-term blood collection in mice.

Sleep-related symptoms in multiple system atrophy: determinants and impact on disease severity / 中华医学杂志(英文版)

BACKGROUND@#Sleep disorders are common but under-researched symptoms in patients with multiple system atrophy (MSA). We investigated the frequency and factors associated with sleep-related symptoms in patients with MSA and the impact of sleep disturbances on disease severity.@*METHODS@#This cross-sectional study involved 165 patients with MSA. Three sleep-related symptoms, namely Parkinson's disease (PD)-related sleep problems (PD-SP), excessive daytime sleepiness (EDS), and rapid eye movement sleep behavior disorder (RBD), were evaluated using the PD Sleep Scale-2 (PDSS-2), Epworth Sleepiness Scale (ESS), and RBD Screening Questionnaire (RBDSQ), respectively. Disease severity was evaluated using the Unified MSA Rating Scale (UMSARS).@*RESULTS@#The frequency of PD-SP (PDSS-2 score of ≥18), EDS (ESS score of ≥10), and RBD (RBDSQ score of ≥5) in patients with MSA was 18.8%, 27.3%, and 49.7%, respectively. The frequency of coexistence of all three sleep-related symptoms was 7.3%. Compared with the cerebellar subtype of MSA (MSA-C), the parkinsonism subtype of MSA (MSA-P) was associated with a higher frequency of PD-SP and EDS, but not of RBD. Binary logistic regression revealed that the MSA-P subtype, a higher total UMSARS score, and anxiety were associated with PD-SP; that male sex, a higher total UMSARS score, the MSA-P subtype, and fatigue were associated with EDS; and that male sex, a higher total UMSARS score, and autonomic onset were associated with RBD in patients with MSA. Stepwise linear regression showed that the number of sleep-related symptoms (PD-SP, EDS, and RBD), disease duration, depression, fatigue, and total Montreal Cognitive Assessment score were predictors of disease severity in patients with MSA.@*CONCLUSIONS@#Sleep-related disorders were associated with both MSA subtypes and the severity of disease in patients with MSA, indicating that sleep disorders may reflect the distribution and degree of dopaminergic/non-dopaminergic neuron degeneration in MSA.

Feasibility of echocardiography-guided repeated intraventricular blood sampling in mice / 中华心血管病杂志

Objective: To investigate the feasibility of echocardiography-guided closed-chest repeated intraventricular blood sampling in mice, and to clarify the maximum blood volume that can be collected by this method, and whether the method can be used for long-term repeated blood collection in mice. Methods: Twenty-four male C57BL/6J mice (10-14 weeks old) were divided into the terminal experiment group (n=4, for investigating the maximum blood amount that could be sampled at one time), the repeated 0.5 ml blood collection group (n=10, sampling 0.5 ml whole blood each time, once every two days for consecutive 4 weeks), and the repeated 0.75 ml blood collection group (n=10, sampling 0.75 ml whole blood each time, once every two days for consecutive 4 weeks). High-frequency echocardiography was used to display the largest section of the left ventricle, guiding the insulin syringe needle through the thorax into the left ventricle for blood collection. In the repeated 0.5 ml blood collection group, echocardiography was used to detect the cardiac structure and function before blood collection, three minutes after blood collection, and one week after the last (the 14th) blood collection. Results: We successfully performed echocardiography-guided closed-chest intraventricular blood sampling, with an average operating time (88±19)s per mouse, and a maximum blood volume (1.43±0.11)ml per mouse. In the repeated 0.5 ml blood collection group, heart rate, left ventricular ejection fraction, left ventricular fractional shortening, left ventricular end-diastolic dimension and left ventricular posterior wall end-diastolic thickness remained uncganged before the first blood collection and after 4 weeks of repeated blood collection (all P>0.05). No death in the repeated 0.5 ml blood collection group. However, in the 0.75 ml blood collection group, two mice died before the end point. Conclusions: The echocardiography-guided closed-chest intraventricular blood sampling is a safe, minimally invasive, convenient and efficient method, and can be used repeatedly for long-term blood collection in mice.

Integrated chemical and transcriptomic analyses unveils synthetic characteristics of different medicinal root parts of Angelica sinensis / 中草药·英文版

Objective: Why are different medicinal parts including heads, bodies and tails of Angelicae Sinensis Radix (ASR) distinct in pharmaceutical activities? Here we explored their discrepancy in chemical constituents and transcriptome. Methods: ASR were separated into three medicinal parts: heads (rootstocks with petiole traces of ASR), bodies (taproots of ASR) and tails (lateral roots of ASR), and chemical and transcriptomic analyses were conducted simultaneously. Results: High performance liquid chromatography (HPLC) fingerprint results showed that five widely used active ingredients (ferulic acid, senkyunolide H, senkyunolide A, n-butylphathlide, and ligustilide) were distributed unevenly in the three ASR medicinal parts. Partial least squares-discriminant analysis (PLS-DA) demonstrated that the heads can be differentiated from the two other root parts due to different amounts of the main components. However, the content of ferulic acid (a main quality marker) was significantly higher in tails than in the heads and bodies. The transcriptome analysis found that 25,062, 10,148 and 29,504 unigenes were specifically expressed in the heads, bodies and tails, respectively. WGCNA analysis identified 17 co-expression modules, which were constructed from the 19,198 genes in the nine samples of ASR. Additionally, we identified 28 unigenes involved in two phenylpropanoid biosynthesis (PB) pathways about ferulic acid metabolism pathways, of which 17 unigenes (60.7%) in the PB pathway were highly expressed in the tails. The expression levels of PAL, C3H, and CQT transcripts were significantly higher in the tails than in other root parts. RT-qPCR analysis confirmed that PAL, C3H, and CQT genes were predominantly expressed in the tail parts, especially PAL, whose expression was more than doubled as compared with that in other root parts. Conclusion: Chemical and transcriptomic analyses revealed the distribution contents and pivotal transcripts of the ferulic acid biosynthesis-related pathways. The spatial gene expression pattern partially explained the discrepancy of integral medicinal activities of three medicinal root parts.