ObjectiveTo explore the main mechanism of self-precipitation formed during the decoction of Sinitang（SNT）， and to provide a research basis for exploring the differences in the toxic and effective components of this compound. MethodsThe average precipitation yields of SNT， Glycyrrhizae Radix et Rhizoma（GRR）-Aconiti Lateralis Radix Praeparata（ALRP） decoction（GF）， ALRP-Zingiberis Rhizoma（ZR） decoction（FJ）， GRR-ZR decoction（GJD）， ALRP decoction（FZ）， ZR decoction（GJ） and GRR decoction（GC） were determined. The four main self-precipitation samples of SNT， GF， FZ and GC were physically characterized by particle size， scanning electron microscopy（SEM）， pH， total dissolved solids（TDS）， conductivity， and Fourier transform infrared spectroscopy（FT-IR） analysis. The chemical compositions of SNT decoction and its different phases was identified by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS） for SNT， SNT self-precipitation and SNT supernatant， and the contents of its main toxic and effective components were determined by high performance liquid chromatography（HPLC）. ResultsPrecipitation yield results of the 7 samples of SNT decoction and single decoction showed that SNT had the highest self-precipitation yield. The formation of SNT self-precipitation was mainly related to the reaction between ALRP and GRR components to form complexes， and FT-IR showed that GRR had the greatest influence on the formation of self-precipitation. A total of 110 components were identified in the SNT decoction， including 100 components in the SNT self-precipitation and 106 components in the SNT supernatant. And quantitative results of the main toxic and effective components revealed that the reaction between ALRP and GRR components formed complexes， resulting in the following content hierarchy for free components：SNT decoctionsupernatantself-precipitation， these components included free liquiritin， benzoylmesaconine， benzoylaconitine， benzoylhypacoitine， liquiritigenin， aconitine， hypoaconitine， isoliquiritigenin and ammonium glycyrrhizinate. ConclusionSNT exhibits spontaneous precipitation during compound decoction， with GRR exerting the greatest influence on its formation. This suggests GRR plays a significant role in the detoxification of SNT. The differences in the self-precipitated toxic-effective components of SNT compound decoction primarily manifest as changes in component content， reflecting the characteristics of SNT "deposition in vitro and sustained release in vivo" and the importance of "administered at draught" in the clinical application of SNT.

Objective: To explore the pattern of change of axial length/curvatrue radius ratio (AL/CR) and associated factors in primary and secondary school students in Yunnan, so as to provide scientific basis and prospective guidance for early myopia intervention and control. Methods: A total of 685 students from grades 2 to 3 and grade 7 in 2 cities/counties in Yunnan Province were selected by multi stage stratified random cluster sampling method in 2020. All the participants were followed up twice with questionnaire of myopia related factors, uncorrected distance visual acuity, and refractive parameter measurement from October,2021 and March,2023,respectivelty. The distribution and change of AL/CR in different classes and groups were analyzed, and the influencing factos of AL/CR cumulative progression were explored using generalized linear model. Results: AL/CR ratio in primary school students was (2.95±0.09) at baseline, increased to (2.99±0.11) at the first follow up and (3.04±0.12) at the second follow up. AL/CR ratio in middle school students(3.08±0.13) at baseline, increased to (3.12±0.15) at the first follow up and (3.15±0.14) at the second follow up. The generalized linear model showed that after controlling for environmental factors, ethnicity ( β =-0.017) , cumulative progression of the SE ( β =-0.027) influenced the changes of AL/CR ratio among the primary school students, whereas the changes of AL/CR ratio were associated with baseline AL ( β =-0.005), baseline corneal curvatrue radius ( β =0.032) and cumulative progression of SE( β =-0.035) among middle school students ( P <0.05). Conclusions The AL/CR ratio of primary and secondary school students in Yunnan can be used to judge different refractive status types, but its variation is not only related to SE progression, but also affected by different ethnic groups and baseline ocular biological parameters. so the value of AL/CR application in assessing the progression of myopia needs to be further confirmed.

BACKGROUND: Intrauterine growth restriction (IUGR) is associated with adverse metabolic outcomes during adulthood. Histone modifications and changes in DNA methylation-affected genes are important for fetal development. This study aimed to confirm the epigenetic mechanisms in IUGR. METHODS: IUGR models were established in Sprague-Dawley rats using a maternal nutritional restriction approach during pregnancy. The abundance of insulin-like growth factor 2 (IGF2), phosphoinositide 3-kinase (PI3K), AKT serine/threonine kinase 2 (AKT2), and PPAR gamma coactivator 1 alpha (PGC-1α) was examined by real-time polymerase chain reaction (RT-PCR) and Western blotting analysis. Chromatin immunoprecipitation RT-PCR was employed to analyze histone modification in CCCTC-binding factor (CTCF)1-4 binding sites of the IGF2/H19 imprinting control region (ICR). The methylation states of CTCF1-4 binding sites were studied by pyrosequencing. RESULTS: The IUGR models were constructed successfully. IGF2 mRNA abundance in the placenta, fetal liver, and newborn liver was decreased in the IUGR group (P <0.01). Meanwhile, as compared with the control group, the expression levels of AKT2, PI3K, and PGC-1α were lower in newborn and 8-week-old livers in the IUGR group (P <0.05). In addition, knocking down IGF2 reduced the protein expression levels of AKT2-P and PGC-1α (P <0.05). In CTCF binding sites 1-4 of the IGF2/H19 ICR, AcH3 enrichment was significantly lower in CTCF1-3 in newborn and 8-week-old IUGR rats. H3K4me3 enrichment was significantly lower in the CTCF1-4 of newborn and 8-week-old IUGR groups (P <0.01). H3K9me2 enrichment was significantly higher in the IUGR group (P <0.01). The CpG dinucleotide methylation levels of CTCF1 and CTCF3, but not those of CTCF2 and CTCF4 binding sites in IUGR rat fetal, 4-week old, and 8-week-old livers decreased significantly (P <0.05). CONCLUSION The methylation status and histone modification in the IGF2/H19 ICR are related to growth and lipid metabolism via the PGC-1α/PI3K/AKT2 pathway in IUGR rats.

The outbreak of Coronavirus Disease 2019 (COVID-19) caused by 2019 novel coronavirus (2019-nCoV) has caused serious public health problems worldwide. Variants of 2019-nCoV with varied virulence and transmissibility have caught attention continuously. Fast, accurate and effective detection methods play a crucial role in the control and prevention of COVID-19 pandemic. Clustered regularly interspaced short palindromic repeats (CRISPR) and related proteins (CRISPR-associated, Cas) based nucleic acid detection methods are characteristic with high specificity and sensitivity, speediness, portability and low price. They have the potential as a candidate one to be used in the molecular detection of 2019-nCoV. The application of CRISPR/Cas system in the detection of 2019-nCoV was reviewed here as a reference for the control and prevention of COVID-19.

Objective:To develop a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats(CRISPR)/Cas12a-based nucleic acid assay for monkeypox virus with high specificity and sensitivity.Methods:RAA primers and CRISPR RNA (crRNA) were designed based on the known conserved regions of the monkeypox virus gene and synthesized, and specific crRNAs were screened using fluorescence detection. The sensitivity and specificity of the detection system were evaluated.Results:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was developed with a sensitivity of 2.5 copies/reaction and high specificity without cross-reactivity with ectromelia virus and vaccinia virus.Conclusions:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was established, which would provide a powerful tool for efficient, rapid and specific detection of monkeypox virus.

Brain natriuretic peptide (BNP), a natriuretic peptide hormone, is mainly secreted by cardiomyocytes when they are pulled by mechanical force. BNP acts to increase natriuresis and decrease vascular resistance, thereby decreasing blood volume, systemic blood pressure and afterload. It is not only the main indicators of cardiac function which is widely used in clinical, recent studies have also shown that it has a very important value in identifying sudden cardiac death(SCD). However, the exact expression of the regulation mechanism is still unknown. This paper reviewed the structure, expression of the regulation mechanism of BNP, and the application progress in clinical and forensic.

Objective T o observe the expression changes of hypoxia inducible factor-1α (H IF-1α) and vascular endothelial grow th factor-A (V E G F-A ) in rats w ith arrhythm ias, and to explore the differences of the expression pattern in the tw o indicators of acute m yocardial ischem ia caused by arrhythm ias and coronary insufficiency. Methods T he arrhythm ia w as induced by C aC l2, and the expression changes of H IF-1α and V E G F-A w ere detected by im m unohistochem istry, W estern blotting and real-tim e PC R w ithin 6 h after the arrhythm ia in rats. Results T he expression of H IF-1α and V E G F-A show ed diffuse in the m yocardial tissue of rats died from arrhythm ias. B oth of them increased in the early arrhythm ia, then decreased. E xtensive m yocardial ischem ia happened at the beginning of arrhythm ia occurrence and its range didn't expand w ith tim e. Conclusion T he expressions of H IF-1α and V E G F-A in m yocardium of the rats w ith arrhythm ia can provide evidence for the differential diagnosis of acute m yocardial is-chem ia caused by fatal arrhythm ia and coronary insufficiency.

Objective To test and eva1uate the abi1ity of three potential chloroplast DNA (cpDNA) barcoding sequences;To find new methods to identify the species of gardenia. Methods Three cpDNA sequences were amplified and sequenced by universal primers of matK, rbcL and psbA. By comparing PCR amplification efficiency, length, intra-and inter-specific divergence, and barcoding gap, BLAST and DNA MAN were used to evaluate these loci. Results The amplification efficiency of 5 samples from 3 gardenia species was 100%. Analysis of the intra-and inter-specific divergence of matK among the sequences showed that barcoding gap was superior to psbA and rbcL, with higher identification efficiency. Conclusion Gardenia jasminoides Ellis can be better identified by matK sequence.

Objective To explore the changes of cell proliferation and differentiation of neural stem cells induced by fasudil treatment,and to primarily study the mechanism in C17.2 mice.Methods C17.2 cells were cultured in vitro; 5,25,50 and 100 μmol/L fasudil were given to the cells,respectively,for 24 h,and cells in the blank control group were given the same volume of culture medium.The changes of cell morphology were observed under a phase-contrast microscope; cell viability and cell necrosis rate were determined by MTT assay and lactate dehydrogenase (LDH) assay,respectively.Western blotting was applied to detect the expression levels of neural markers (nestin,glial fibrillary acidic protein [GFAP],double cortisol [DCX],microtubule-associated protein-2 [MAP-2]),and Notch Ⅰ and Hes 1 proteins in the notch signaling in cells from the 100 μmol/L fasudil treatment group and blank control group.Immunofluorescence staining was used to detect the nestin and GFAP expressions in the C 17.2 cells.Results As compared with that in the blank control group,the cell viability in the 50 and 100 μmol/L fasudil treatment groups was significantly decreased; that in the 100 μmol/L fasudil treatment group was significantly lower than that in 50 μmol/L fasudil treatment group (P＜0.05); LDH assay showed no significant difference of cell necrosis among the five groups (P＜0.05).Western blotting indicated that 100 μmol/L fasudil treatment group had significantly decreased nestin expression,significantly elevated DCX,MAP-2 and GFAP expressions,and statistically decreased expression levels of Notch 1 and Hes 1 as compared with blank control group (P＜0.05).Immunofluorescence staining indicated that the percentage of nestin positive cells was markedly decreased,the percentage of GFAP positive cells was significantly increased in the 100 μmol/L fasudil treatment group as compared with those in the blank control group (P＜0.05).Conclusion Fasudil treatment could inhibit the proliferation of C17.2 cells and promote them differentiate into neuronal and glial cells via decreasing the expression level of Notch signaling.