Chronic glomerulonephritis （CGN） is a common clinical chronic glomerular disease caused by autoimmune reaction， the pathogenesis of which is complex and has not been fully elucidated. There is no specific treatment method in modern medicine. The establishment of an animal model of CGN in accordance with its characteristics in western medicine and traditional Chinese medicine will help to reveal the pathogenesis of CGN， rate drugs， and improve the treatment plan. Based on the clinical diagnostic criteria of CGN， the paper establishes the syndrome differentiation criteria of CGN for Chinese and western medicine. Through summarizing the literature on animal models of CGN and making a further analysis， it is found that the CGN models are mainly modeled using rats with the methods of single-factor induction or two-factor induction， and the main manifestation of the disease characteristics is nephritis-related symptoms. The single-factor-induced or two-factor-induced CGN rat models have a high fitting degree with the clinical characteristics in western medicine， but the fitting degree is insufficient with the clinical characteristics in traditional Chinese medicine. In addition， the CGN models with syndromes of traditional Chinese medicine are dominated by Qi deficiency in the spleen and kidney and Qi deficiency in the lung and kidney， while models for Yang deficiency in the spleen and kidney， Yin deficiency in the liver and kidney， and deficiency of both Qi and Yin are slightly insufficient. Therefore， it is important to prepare a new and improved animal model of CGN， so that a preclinical model can be provided for the exploration of the pathogenesis of CGN in western medicine and traditional Chinese medicine and its therapeutic research.

Objective To explore the effect of open reading frame 66(C12ORF66)located at chromosome 12 on the viability of MYCN amplified NB cell lines.Methods DDatasets GSE16476 and GSE49710 in R2 database were analyzed for expression level of C12ORF66 in MYCN amplified and MYCN non-amplified NB cells and its potential correlation with the prognosis of pediatric patients.C12ORF66 mRNA expression level in normal tissue immortalized cell lines,MYCN amplified and MYCN non-amplified cell lines were detected by RT-qRCR.Transient or stable knockdown of C12ORF66 cell lines were constructed to compare the difference in real time cellular analysis(RTCA),colony formation,Ki67 positive cells between the control group and the C12ORF66 knockdown group.Results By analyzing R2 datasets,C12ORF66 level in MYCN amplified samples was significantly higher than that in MYCN non-amplified samples,and the expression of C12ORF66 was negatively correlated with the prognosis of pediatric patients(P＜0.05).C12ORF66 highly expressed in MYCN-amplified BE(2)-C and SK-N-BE(2)cell lines than in MYCN non-amplified CHLA-255 and SH-SY5Y cell lines(P＜0.001).Transient or stable knockdown of C12ORF66 resulted in significant slow down of proliferation of MYCN amplified NB cells(P＜0.001),the colony formation ability was significantly reduced(P＜0.001),and the proportion of Ki67 positive cells was significantly decreased(P＜0.05).Conclusions C12ORF66 was highly expressed in MYCN amplified clinical NB samples and cell lines which is believed to be correlated with poor prognosis of pediatric patients.C12ORF66 knockdown signifi-cantly inhibits cell viability of NB cells.

ObjectiveThe human angiotensin converting enzyme2 （hACE2） transgenic mouse model was used to clarify the antiviral efficacy of BD-77 against a novel coronavirus SARS-CoV-2 and explore the action mechanism of BD-77 against SARS-CoV-2. MethodSARS-CoV-2 Omicron and Delta variant strains-infected VeroE6 cell models were established and administered with BD-77 to observe the antiviral effect of BD-77 in vitro. A kit was used to detect the effect of BD-77 in vitro on the binding of spike S protein of SARS-CoV-2 virus （Delta/Omicron） to angiotensin converting enzyme2 （ACE2）. Chromatography was adopted to detect the binding of BD-77 to the S protein and N protein of the novel coronavirus. hACE2 transgenic C57BL/6 mice were divided into a blank control group， SARS-CoV-2 infection group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1， with eight mice in each group. The pneumonia model of SARS-CoV-2-infected hACE2 transgenic mice was built to observe the survival of the mice， detect the virus titer of the lung tissue of the mice， and observe the lesions in the lung tissue. ResultBD-77 had a certain inhibitory effect on Omicron and Delta variant strains in vitro， with median inhibitory concentration （IC₅₀） of 526.3 mg·L^-1 and 653.0 mg·L^-1， respectively. BD-77 had no significant inhibitory effect on the binding of the S protein of WT， Omicron， and Delta variant strains of SARS-CoV-2 to ACE2 and had no binding effect with the S protein and N protein of the novel coronavirus. No mice in the blank group died， while the mortality rate of SARS-CoV-2-infected mice was 75%. There was a large amount of virus replication in the lung tissue of the mice and large areas of inflammatory infiltration in the lung tissue and interstitium. Compared with the model group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1 could reduce the mortality of mice， significantly lower the virus titer in the lung tissue of mice （P<0.05）， and improve lung lesions. ConclusionBD-77 demonstrated significant inhibitory effects against SARS-CoV-2 virus in vitro and in vivo. However， its mechanism of action did not involve direct inhibition of the virus itself or intervention in the virus-host binding process. This finding suggests that the mechanism of action of BD-77 needs to be thoroughly investigated and elucidated by further experiments.

Objective:To investigate the function and underlying mechanism of cysteine and glycine-rich protein 2(CSRP2)in neuroblastoma(NB).Methods:The correlation between the expression level of CSRP2 mRNA and the prognosis of NB children in NB clinical samples was analyzed in R2 Genomics Analysis and Visualization Platform.The small interfering RNA(siRNA)targeting CSRP2 or CSRP2 plasmid were transfected to NB cell lines SK-N-BE(2)and SH-SY5Y.Cell proliferation was observed by crystal violet staining and real-time cellular analysis.The ability of colony formation of NB cells was ob-served by colony-forming unit assay.Immunofluorescence assay was used to detect the expression of the proliferation marker Ki-67.Flow cytometry analysis for cell cycle proportion was used with cells stained by propidium iodide(PI).Annexin V/7AAD was used to stain cells and analyze the percentage of cell apoptosis.The ability of cell migration was determined by cell wound-healing assay.The level of protein and mRNA expression of CSRP2 in NB primary tumor and NB cell lines were detected by Western blot and quantitative real-time PCR(RT-qPCR).Results:By analyzing the NB clinical sample databases,it was found that the expression levels of CSRP2 in high-risk NB with 3/4 stages in international neuroblas-toma staging system(INSS)were significantly higher than that in low-risk NB with 1/2 INSS stages.The NB patients with high expression levels of CSRP2 were shown lower overall survival rate than those with low expression levels of CSRP2.We detected the protein levels of CSRP2 in the NB samples by Western blot,and found that the protein level of CSRP2 in 3/4 INSS stages was significantly higher than that in 1/2 INSS stages.Knockdown of CSRP2 inhibited cell viability and proliferation of NB cells.Overexpression of CSRP2 increased the proliferation of NB cells.Flow cytometry showed that the proportion of sub-G1,G0/G1 and S phase cells and Annexin V positive cells were increased after CSRP2 deficiency.In the cell wound-healing assay,the healing rate of NB cells was significantly attenuated after knockdown of CSRP2.Further mechanism studies showed that the proportion of the proliferation marker Ki-67 and the phospho-rylation levels of extracellular signal-regulated kinases 1/2(ERK1/2)were significantly decreased after CSRP2 knockdown.Conclusion:CSRP2 is highly expressed in high-risk NB with 3/4 INSS stages,and the expression levels of CSRP2 are negatively correlated with the overall survival of NB patients.CSRP2 significantly increased the proliferation and cell migration of NB cells and inhibited cell apoptosis via the activation of ERK1/2.All these results indicate that CSRP2 promotes the progression of NB by activating ERK1/2,and this study will provide a potential target for high-risk NB therapy.

The observational research based on big data in healthcare has attracted increasing attention, with the control and evaluation of residual confounding being the critical issue that needs to be solved urgently. This review summarized the methods for statistical adjustment and sensitivity analysis of residual confounding in the association analysis with a multicenter database. Based on individual-level data, the residual confounding can be adjusted in each subcenter using methods such as regression discontinuity design, while the pooled estimate can be obtained as a weighted average. Based on the center-level results, the Bayesian Meta-analysis method can adjust the pooled estimates. The sensitivity analysis of residual confounding can also be carried out using center-level data to calculate the E-value, p^(q), T^(r,?q) and G^r,?q. The abovementioned methods should be selected reasonably according to the requirements for practical applications, advantages, and disadvantages. For example, the use of subcenter individual data for residual confounding adjustment usually needs strict study design and frequent coordination; the Bayesian Meta-analysis is based on some strong assumptions; the interpretation of the results in the sensitivity analysis, such as E-value requires professional judgment to assess the risk of residual confounding. Therefore, the methods for controlling and evaluating residual confounding in association analysis based on multicenter databases still need further development and improvement.

ObjectiveTo study the changes of dry matter accumulation， mineral element absorption and medicinal component accumulation in Belamcanda chinensis and to provide a theoretical basis for fertilization in the standardized cultivation of B. chinensis. MethodDuring the two-year growth period of B. chinensis， the samples were collected at 30-day intervals from day 90 after sowing for the measurement of dry matter accumulation and mineral element content in different organs as well as the flavonoid content in the rhizome. ResultAccording to the accumulation of dry matter in different organs， the two-year growth period of B. chinensis was divided into seven stages： seedling stage （before July in the first year）， first-year vegetative growth stage （August to October in the first year）， first-year late growth stage （November to January of the second year）， re-growth stage （February to March in the second year）， second-year vegetative growth stage （April to May in the second year）， reproductive growth stage （June to October in the second year）， second-year late growth stage （November of the second year to January of the third year）. During the whole growth period， the late growth stage of each year witnessed the expansion of the rhizome， and the accumulation of dry matter of the whole plant peaked after the reproductive growth stage. The content of flavonoids in the rhizome varied greatly among different stages. Specifically， the content of mangiferin， tectorigenin and irigenin was higher in the second-year vegetative growth stage， and that of tectoridin and iridin was higher in the first-year late growth stage and the second-year late growth stage. The content of irisflorentin was higher in the second-year vegetative growth stage and the second-year late growth stage than that in other stages. The total content of six flavonoids peaked in the second-year vegetative growth stage （April in the second year）. The correlation analysis showed that all the nine mineral elements had different degrees of positive correlations with tectorigenin， irigenin and irisflorentin. ConclusionIn the first year， B. chinensis presents slow dry matter accumulation and low demand for fertilizer， so the application of base fertilizer can be reduced. In the second year， especially after the reproductive growth， B. chinensis demands high fertilizer， so nitrogen， phosphorus and potassium fertilizers and other micronutrients should be applied as early as possible. In addition， the rhizome expands in October to November of the first year and November to December of the second year， when the supply of nitrogen and potassium to the rhizome should be ensured or beneficial elements should be supplemented to improve the quality of B. chinensis.

ObjectiveTo investigate the effects of different nitrogen application rates on the growth， yield and quality of Belamcanda chinensis and to provide a theoretical basis for the quality and production efficiency improvement of B. chinensis. MethodA field experiment was carried out on the basis of 120 kg·hm^-2 phosphate and potassium fertilizers， with five nitrogen fertilizer treatments： 0 （N₀）， 60 （N₆₀）， 120 （N₁₂₀）， 180 （N₁₈₀） and 240 （N₂₄₀） kg·hm^-2. The effects of different nitrogen application rates on the agronomic traits， yield， mineral element accumulation and flavonoid content of B. chinensis were studied. ResultApplication of nitrogen fertilizer was beneficial to dry matter accumulation and significantly increased the yield of rhizome. After the application of nitrogen fertilizer， the yield of rhizome increased by 48.75%-86.62%. The nitrogen application at 60-180 kg·hm^-2 increased the active components such as tectoridin， iridin and irisflorentin， while it was not conducive to the accumulation of tectorigenin and irigenin in the rhizome. The accumulation of Mn and Fe reached the highest when the nitrogen application rate was 120 and 60 kg·hm^-2， respectively. The principal component analysis of active component content showed that the comprehensive score was ranked as N₁₈₀>N₁₂₀>N₆₀>N₂₄₀>N₀. The correlation analysis indicated that the tectoridin content had a positive correlation with N accumulation and a negative correlation with Ca accumulation in B. chinensis. The content of tectorigenin was negatively correlated with N accumulation and positively correlated with Ca accumulation. The content of irigenin had positive correlations with Ca and Zn accumulation. ConclusionThe nitrogen application at 120-180 kg·hm^-2 can increase the rhizome yield and the content of tectoridin， iridin and irisflorentin in the rhizome， so as to improve the yield and quality of this herb.

ObjectiveTo investigate the effect of the combined application of organic fertilizer and chemical fertilizer at different ratios on the growth， yield and quality of two-year-old Belamcanda chinensis and provide a scientific basis for fertilization to achieve high yield and quality of this medicinal plant. MethodIn a field plot experiment， we designed five fertilization patterns with organic fertilizer and chemical fertilizer at different ratios： OM₀ （pure chemical fertilizer）， OM₁ （30% organic fertilizer + chemical fertilizer， application of organic fertilizer at 2 567 kg·hm^-2）， OM₂ （50% organic fertilizer + chemical fertilizer， application of organic fertilizer at 4 283 kg·hm^-2）， OM₃ （70% organic fertilizer + chemical fertilizer， application of organic fertilizer at 6 000 kg·hm^-2）， and OM₄ （pure organic fertilizer， 8 567 kg·hm^-2）. The effect of different fertilization patterns on the agronomic properties， herb yield， flavonoid content， mineral element content， and fertilizer utilization rate of B. chinensis were determined. ResultThe combined application of organic fertilizer and chemical fertilizer at an appropriate ratio increased the leaf width and promoted tillering， thereby improving the yield of B. chinensis. The yield of OM₁ and OM₂ groups increased by 6.65% and 13.10%， respectively， compared with that of OM₀ group， while too high a proportion of organic fertilizer led to a decrease in yield. With organic fertilizer application， the content of mangiferin， tectoridin， iridin， tectorigenin， irigenin， and irisflorentin in the rhizomes of B. chinensis increased. Moreover， a certain proportion of organic fertilizer facilitated the accumulation of functional elements such as K， Fe and Cu. The results of the principal component analysis showed that the overall quality of B. chinensis was ranked in the order of OM₃ > OM₂ > OM₁ > OM₄ > OM₀. The OM₂ group had the maximum accumulation of N， P and K and the maximum partial factor productivity of fertilizer. ConclusionAccording to the agronomic traits， yield， quality indexes and fertilizer utilization rate of B. chinensis under different fertilization patterns， we recommended to apply 4 283 kg·hm^-2 organic fertilizer in combination with chemical fertilizer to the production of two-year-old B. chinensis， so as to improve the quality and yield of B. chinensis.

ObjectiveTo investigate the effects of different phosphorus application rates on the growth， yield and quality of two-year-old Belamcanda chinensis and provide a basis for phosphorus application in the high-quality and high-yielding cultivation of B. chinensis. MethodIn this study， we designed five phosphorus application rates of 0 （P₀）， 60 （P₆₀）， 120 （P₁₂₀）， 180 （P₁₈₀） and 240 （P₂₄₀） kg·hm^-2 on the basis of 120 kg·hm^-2 nitrogen and potassium fertilizers to investigate the effects of different phosphorus levels on the agronomic traits， yield， mineral content， flavonoid content and phosphorus utilization efficiency of B. chinensis through field experiments. ResultThe plant height and tillering number of B. chinensis gradually increased after phosphorus fertilization. The phosphorus application increased the rhizome yield by 37.59%-61.03% compared with P₀ treatment and the highest yield was achieved when phosphorus fertilizer was applied at 120 kg·hm^-2. The content of mangiferin and irisflorentin in the rhizomes gradually increased with the increase in phosphorus application and the moderate application of phosphorus fertilizer increased the content of tectoridin， iridin and irigenin. The highest Mn， Cu and Fe elements were achieved at 60 and 120 kg·hm^-2 of phosphorus fertilization， respectively. The overall quality scores of B. chinensis were ranked in the order of P₂₄₀>P₁₈₀>P₁₂₀>P₆₀>P₀. With the increase in phosphorus application， the phosphorus accumulation of B. chinensis first increased and then decreased， and the partial factor productivity， apparent utilization rate and agronomic efficiency of phosphorus fertilizer gradually decreased. ConclusionConsidering the yield， quality and phosphorus utilization efficiency of B. chinensis， it is recommended to apply phosphorus fertilizer at about 120 kg·hm^-2 in the production of B. chinensis in Tuanfeng county.

ObjectiveTo clarify the accumulation and distribution characteristics of dry matter and mineral elements in Artemisia argyi var. argyi cv. Qiai， and to provide technical support for the high yield of and efficient utilization of nutrients in this medicinal species. MethodTwo cultivars of this species， Qiqing 1 and Qihuang 1 were selected， and the composition of dry matter in different organs， the content， accumulation， and distribution of mineral elements in each organ of the two cultivars， and the dynamic changes of volatile oil content and index components eucalyptol and borneol in leaves of the two cultivars were monitored at different growth stages. ResultThe period from February to March marked the early growth stage of Qiai， and the dry matter was mainly distributed in the leaves. It accelerated the growth in April， and the period from April to mid-June witnessed the vigorous vegetative growth of Qiai， during which the dry matter was mainly found in the stems and leaves. It began the reproductive growth from late June and the dry matter was mainly distributed in the stems. In the flowering stage in August， no dry matter accumulation occurred. As for the volatile oil， the content was high （> 1.10%） at the vigorous vegetative growth stage and peaked on June 14 （1.33% in Qiqing 1， and 1.23% in Qihuang 1）. The relative mass fraction of eucalyptol was the maximum at the vegetative growth stage （8.67% in Qiqing 1， and 13.07% in Qihuang 1）. The relative mass fraction of borneol peaked at the early growth stage （2.63% in Qiqing 1， and 5.94% in Qihuang 1）. The content of nitrogen， phosphorus， potassium， and zinc in leaves was in significantly positive correlation with the content of volatile oil and the relative content of eucalyptol and borneol. The content of macroelements nitrogen， phosphorus， potassium， and calcium and trace elements iron and zinc peaked at the early growth stage， and the content was the highest in stem and leaf. The content of macroelement magnesium and trace elements manganese and copper was the highest at vegetative growth stage when the content of other elements decreased and the nutrients were gradually transferred to the buds， flowers and other organs. In the whole growth period， the distribution of potassium， calcium， and zinc was in the order of leaf > stem > root， and the distribution of nitrogen， phosphorus， copper， magnesium， and manganese followed the order of leaf > root > stem. The distribution of iron was in the order of root > leaf > stem. There was a significantly positive correlation between the total amount of dry matter and the absorption of nutrients in 'Qiai'. The absorption of macroelements by Qiai was in the order of potassium > nitrogen > calcium > phosphorus > magnesium， and the ratio of absorbed elements was about 2.66∶2.51∶0.6∶0.11∶0.04. The absorption of trace elements followed the order of manganese > iron > zinc > copper， and the ratio of absorbed elements was about 0.25∶0.17∶0.05∶0.04. In terms of the production of medicinal materials， 'Qiai' needed about 4.11 kg potassium， 3.58 kg nitrogen， 0.91 kg phosphorus， 0.18 kg calcium， 0.06 kg magnesium， about 6.64 g manganese， 2.56 g iron， 1.30 g zinc， and 0.92 g copper to produce 100 kg medicinal materials. ConclusionEnough organic fertilizer and phosphorus and potassium fertilizers should be applied as base fertilizers for Qiai. The vegetative growth stage （April-June） marks the high accumulation of dry matter and large demand of nutrients， during which topdressing should be conducted timely and early， especially nitrogen fertilizer， and appropriate amount of micro-element fertilizer should be added. Qiai needs a large amount of calcium and magnesium fertilizers from the mid-vegetative growth stage， and they should be applied in time in the late stage to ensure the vegetative growth of the plants for seeds and the quality of the medicinal material of Qiai.