ObjectiveThe detection of RNA single nucleotide polymorphism (SNP) is of great importance due to their association with protein expression related to various diseases and drug responses. At present, splintR ligase-assisted methods are important approaches for RNA direct detection, but its specificity will be limited when the fidelity of ligases is not ideal. The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection. MethodsIn this study, a dual-competitive-padlock-probe (DCPLP) assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation. To verify the method, we employed dual competitive padlock probe-mediated rolling circle amplification (DCPLP-RCA) to genotype the CYP2C9 gene. ResultsThe specificity was well improved through the competition and strand displacement of dual padlock probe, with an 83.26% reduction in nonspecific signal. By detecting synthetic RNA samples, the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L. Furthermore, clinical samples were applied to the method to evaluate its performance, and the genotyping results were consistent with those obtained using the qPCR method. ConclusionThis study has successfully established a highly specific direct RNA SNP detection method, and provided a novel avenue for accurate identification of various types of RNAs.

Child ; Humans ; Toxoplasmosis, Cerebral ; Hematopoietic Stem Cell Transplantation ; Thalassemia

Objective:To investigate the effect of early postoperative enteral nutrition(EN)on postoperative rehabilitation and inflammation after laparoscopic radical gastrectomy for gastric cancer,in order to provide reference for postoperative rehabilitation of such patients.Methods:Patients who received laparoscopic assisted radical gastrectomy in Department of Gastrointestinal Surgery of The First Affiliated Hospital of Bengbu Medical College from January 2020 to December 2022 were included in the analysis.According to the different ways of postoperative nutritional treatment,patients were divided into the observation group(early postoperative EN group)and the control group(parenteral nutrition group),and indexes such as postoperative rehabilitation,abdominal drainage flow and the level of inflammatory mediators in drainage fluid were compared between the two groups.Results:A total of 81 patients were included,including 41 in the observation group and 40 in the control group.Interval of the first postoperative exhaust(t=3.806;P＜0.001)and resuming diet day(t=5.510;P＜0.001),and length of postoperative hospital stay(t=2.401;P=0.019)in the observation group were shorter than those in the control group.Levels of peripheral blood albumin(t=14.040;P＜0.001)and prealbumin(t=9.832;P＜0.001)of the observation group at postoperative day(POD)5 were significantly higher than those of the control group,but there was no significant difference in hemoglobin level(t=1.477;P=0.144).The level of CRP in peripheral blood of the observation group at POD 5(t=7.758;P＜0.001)and the incidence of postoperative SIRS[(12.2％,5/41)vs(32.5％,13/40),x2=4.830;P=0.028)]were significantly lower than those in the control group.The average drainage volume(t=6.858;P＜0.001),drainage removal time(t=5.016;P＜0.001),and TNF-α level(t=4.993;P＜0.001)and IL-6 level(t=20.640;P＜0.001)in postoperative drainage at POD 5 were significantly lower in the observation group than those in the control group.Conclusion:Early postoperative EN could accelerate the rehabilitation process after laparoscopic radical gastrectomy,improve postoperative nutritional status,and reduce abdominal inflammation.

Zhenwutang is the classic formula of Wenyang Lishui in " Treatise on Febrile Diseases" . It consists of five kinds of medicines, namely Aconiti Lateralis Radix Praeparata, Zingiberis Rhizoma Recens, Atractylodis Macrocephalae Rhizoma, Poria, Paeoniae Radix Alba or Paeoniae Radix Rubra. It has been included in the Catalogue of Ancient Classics (The First Batch) issued by the State Administration of Traditional Chinese Medicine in 2018.There are two articles in the " Treatise on Febrile Diseases" about the syndromes of Zhenwutang. On the basis of Article 82 of the Chapter of Taiyang disease, the Taiyang disease is demonstrated by sweating, but after that the patient still suffers from fever, epigastric throb, dizziness and shiver, this shall be treated with Zhenwutang. According to Article 316 of the chapter of Shaoyin disease, Shaoyin disease lasts for two or three days, until the fourth and fifth days, and is demonstrated by abdominal pain, difficult urination, severe pain in the limbs and diarrhea, the patient have a cough, diuresis, diarrhea and vomit, this shall be treated with Zhenwutang. The original texts discuss the basic pathogenesis is edema syndrome due to Yin and Yang deficiency, with symptoms of unfavorable urination, heavy limbs or edema, pale tongue, white moss and heavy pulse as dialectical points. In terms of the compatibility of prescriptions, usage and dosage, and the interpretation of prescriptions, this paper examines Zhenwutang syndromes in " Treatise on Febrilediseases" , summarizes the material basic research of Zhenwutang, and provides the basis for the quality control of its compound pharmacodynamics. This study summarizes the progress of pharmacological research for Zhenwutang in recent years, finds that Zhenwutang has many efficacies, such as cardiotonic, diuretic, lowering lipid, anti-oxidation, improving renal function and balancing of water and liquid metabolism, lists traditional Chinese medicine(TCM) syndromes treated with Zhenwutang, such as heart palpitations, edema, cough, abdominal pain, sipping, dizziness, and Western medicine diseases, like congestive heart failure, pulmonary heart disease, bronchial asthma, chronic colitis, cirrhosis, ascites, chronic glomerulonephritis, chronic renal failure, benign prostatic hyperplasia and Meniere's syndrome, expounds the principle of Zhenwutang for " treating different diseases with the same therapy" , in order to provide useful reference for the research and development and clinical application of Zhenwutang and more classic prescriptions.

To investigate the potential toxic effects and mechanisms of Tris(1; 3-dichloro-2-propyl) phosphate (TDCIPP) on thyroid in female SD rats. Thirty-two 3-weeks-old female SD rats were randomly divided into normal group(treated with corn oil ), and low/moderate/high-dose group treated with TDCIPP (dissolved in corn oil )(n=8). All rats were treated with corn oil or TDCIPP (50, 100, 250 mg/(kg·d)) once a day during a 21-day period. All rats were sacrificed after the last administration. Serum thyroid stimulating hormone (TSH), 3,3',5-triiodothyronine (T3), 3,3',5,5'-tetraiodothyronine (T4), free 3,3',5,5'-tetraiodothyronine (FT4) were detected with ELISA kit. Morphology of thyroid was observed with hematoxylin and eosin (HE) staining. Expressions of genes and proteins correlate with thyroid were measured respectively by real-time fluorescence quantitative PCR and Western blot. Compared with control group, morphology of thyroid showed follicles irregular arrangement, hypocolloid, and follicular hyperplasia in TDCIPP treatment groups. The levels of serum TSH in low-dose TDCIPP group and T3 in high-dose TDCIPP group were significantly higher than those in control group(P＜0.05). Thyroid stimulating hormone receptor (TSHR) mRNA expression was decreased distinctly in low-dose TDCIPP group, while the expression of thyroperoxidase (TPO) mRNA was increased notably in moderate and high-dose TDCIPP groups(P＜0.05,P＜0.01). Compared with control group, the level of TRβ protein was decreased significantly in moderate and high-dose TDCIPP groups, while the expressions of udp-glucuronosyl-transferases (UGTs) and cytochrome-p450-3A1 (CYP3A1) proteins were upregulated notably in TDCIPP treatment groups(P＜0.05). Treated with 50 mg/(kg·d) TDCIPP can cause thyroid hyperplasia, change the levels of thyroid hormones, and disturb thyroid function, therefore, it has toxic effects on the thyroid.

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires

Yangjing Zhongyu Tang, from FU Qing-zhu Nvke, is a famous classical formula of clinical value for treating blood deficiency syndrome of female infertility in the Qing dynasty. The prescription seems plain, but it is rigorous and effective with high research value. In this paper, the ancient books and modern documents of Yangjing Zhongyu Tang were analyzed from the aspects of traceability of Chinese materia medica in the formula, pharmacological research, clinical research, etc. It could be concluded that the pharmacological studies of this formula were mostly to investigate the mechanism and efficacy of its treatment for infertility, but there was a lack of comprehensive interpretation of the structure, function and principle of pharmacodynamic substance in this formula. In this paper, combined with the contents of supramolecular imprinting template and network pharmacology, the new direction of pharmacological research of Chinese medicine compound was put forward. Quality control of Yangjing Zhongyu Tang mostly based on small molecule compounds, so it was suggested to break the inherent thinking, and increase the detection of macromolecule compounds and supramolecular of Chinese materia medica. The druggability of this formula involved in the preliminary study of decoctions, plasters and granules. Considering the quality stability of Chinese materia medica in the formula and the scientificity of druggability of this formula, the compatibility principle of traditional Chinese medicine (TCM) and modern supramolecular chemistry theory could be combined to study the change law of druggability of TCM before and after compatibility, so as to provide new reference materials for the follow-up clinical application and development of Yangjing Zhongyu Tang.

Objective: Based on the analysis of the total components of fingerprint and the determination methods of the existing components in the 2015 edition of Chinese Pharmacopoeia, the "point-line-surface" quality standard of Yangjing Zhongyu Tang was established by the "point" of each single component (morroniside, loganin, paeoniflorin, ferulic acid and verbascoside) to the "line" of multicomponent and the "face" of fingerprint of the whole component. Method: XB-C₁₈ column (4.6 mm×250 mm, 5 μm) was used for gradient elution of 0.1% phosphoric acid aqueous solution-acetonitrile. The column temperature was 30℃, the injection volume was 10 μL, the flow rate was 1.0 mL·min^-1, and the detection wavelengths were 240, 316, 230, 334 nm. The contents of these five components in Yangjing Zhongyu Tang were determined by three correction methods, external standard method and regression equation method. At the same time, the fingerprint of Yangjing Zhongyu Tang were analyzed by total component analysis and similarity evaluation. Result: With ferulic acid as reference, the relative correction factor (f) of morroniside, loganin, paeoniflorin and verbascoside were 0.392 1, 0.421 4, 0.261 7, 0.268 6 by multi-point correction method, and their f (slope correction method) were 0.385 4, 0.419 4, 0.255 9, 0.274 0, respectively. Twenty characteristic peaks of fingerprint were analyzed and the similarity was ≥ 0.999.There was no significant difference in the contents of these five components from Yangjing Zhongyu Tang determined by the quantitative assay of multi-components by single-marker (QAMS) correction method, the external standard method and the regression equation method. Conclusion: The comprehensive quality standard established by the total component analysis of fingerprint combined with various determination methods of existing components in the 2015 edition of Chinese Pharmacopoeia has been validated in famous classical formula of Yangjing Zhongyu Tang, which can provide ideas and methods for the quality control with quantitative determination and fingerprint of other famous classical formulas.

OBJECTIVE: To explore the effect of bone morphogenetic protein 4(BMP4) on the cell cycle and apoptosis of hemaropoictic stem and progenitor cells (HSPC) in conditions of 5-fluorouracil (5-FU)-inducing bone marrow suppression and stress hemogenesis, and its possible mechanism. METHODS: The C57BL transgenic mice with BMP4 overexpression were established and were enrolled in transgenic group (BMP4 group), at the same time the wild type mice matching in age, sex and body weight were selected and were enrolled in control group (WT group). The bone marrow suppression was induced by injection with 5-FU in dose of 150 mg/kg, then the nucleated cells were isolated from bone marrow. After the HSPCs were markered with C-kit/sca-1 fluorescent antibodies, the changes of cell cycle and apoptosis of HSPC were detected by Aunexin V/PI and Ki67/DAPI double staining; the cell cycle-essociated hemotopoietic regulatory factors were detected by RT-qPCR. RESULTS: Under physiologic status, there were no significant differences in cell cycle and apoptotic rate of HSPC between WT group and BMP-4 group. After the bone marrow was suppressed, the ratio of HSPC at G0 phase in BMP4 group significantly decreased(P＜0.05); the apoptosis rate of HSPC significantly increased(P＜0.05); the mRNA expression levels of hypoxia-inducing factor Hif-1α and chemotactic factor CXCL12 in stroma of BMP4 group were down-regulated significanfly(P＜0.05). CONCLUSION Under non-physiologic conditions such as stress hemogenesis or bone marrow suppression, the up-regulation of BMP4 can promote HSPC into cell cycle and apoptosis of HSPC, moreover, the BMP4 may play a regulatory role for cell cycle of HSPC through direct or indirect down-regulation of Hif-1α and CXCL-12 expressions.
Animals ; Antineoplastic Agents ; Apoptosis ; Bone Morphogenetic Protein 4 ; Cell Cycle ; Hematopoietic Stem Cells ; Mice ; Mice, Inbred C57BL

OBJECTIVETo investigate the effect of propofol and operative trauma on the neurodevelopment and cognitive function of the developing brain and its mechanism.

METHODSA total of 104 postnatal day 13 Sprague-Dawley rats were randomly divided into 4 groups: control group (treated by 7.5 mL/kg saline and sham surgery), propofol group (treated by 75 mg/kg propofol), surgery group (with abdominal surgery under local anesthesia) and propofol+surgery group (with abdominal surgery under local anesthesia plus 75 mg/kg propofol anesthesia). Thirteen rats from each group were randomly selected for detecting the content of TNF-α in the hippocampus and the expression levels of caspase-3 and c-fos in the brain. Morris Water Maze test was used to detect the cognitive ability of the other rats at 60 days old, after which TNF-α content in the hippocampus and caspase-3 and c-fos expressions in the brain were detected.

RESULTSIn 13 day-old rats, TNF-α level and caspase-3 and c-fos expressions differed significantly between the surgery group and the other 3 groups (P<0.05) and were similar among the control group, propofol group and propofol+surgery group (P>0.05). In 60-day-old rats, Morris water maze test results, TNF-α level or expressions of caspase-3 and c-fos showed no significant differences among the 4 groups.

CONCLUSIONAbdominal surgery can induce inflammation in the hippocampus and neuroapoptosis in neonatal rats rather than adult rats. Single-dose propofol anesthesia does not significantly affect neurodevelopment of young rats, and can relieve central inflammatory reaction induced by surgical trauma.