ObjectiveTo observe the effect of M1 bone marrow-derived macrophages （M1-BMDM） with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis， and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDM^Wnt5a-KD cells. A rat model of liver cirrhosis induced by CCl₄/2-AAF was established， and at the end of week 8， rats were randomly divided into model group， M1-BMDM group， M1-BMDM Wnt5a-knockdown empty vector group （M1-BMDM^KD-EV group）， and M1-BMDM Wnt5a-knockdown group （M1-BMDM^Wnt5a-KD group）， with 6 rats in each group. On the first day of week 9， the rats in each group were given a single injection of the corresponding cells via the caudal vein， along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology， serum liver function parameters， hepatic stellate cell activation， and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase （AST） in serum （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group （P<0.05）. The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group， all cell treatment groups had a significant reduction in the percentage of CD68-positive area （all P<0.05）， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area （both P<0.05）. Compared with the model group， all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α （all P<0.05） and the protein expression level of CD68 （all P<0.01）； compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 （both P<0.05）， significant reductions in the protein and mRNA expression levels of CD68 （both P<0.05）， and a significant reduction in the protein expression level of tumor necrosis factor-α （P<0.01）. Sirius Red collagen staining and alpha-smooth muscle actin （α-SMA） immunohistochemical staining showed that compared with the model group， all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area， with the most significant changes in the M1-BMDM^Wnt5a-KD group， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue （all P<0.05）. Compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β （all P<0.05）. Compared with the model group， all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue （all P<0.05）， and the M1-BMDM^Wnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDM^KD-EV group （both P<0.05）. The M1-BMDM^Wnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDM^KD-EV group （P<0.01）. Immunofluorescence co-staining showed that compared with the model group， all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly higher number of such cells than the M1-BMDM^KD-EV group （P<0.05）. ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl₄/2-AAF， which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.

ObjectiveBased on traditional quality evaluation of Gardeniae Fructus（GF） recorded in historical materia medica， this study systematically compared the quality differences between wild and cultivated GF from morphological characteristics， microscopic features， and contents of primary and secondary metabolites. MethodsVernier calipers and analytical balances were used to measure the length， diameter and individual fruit weight of wild and cultivated GF， and the aspect ratio was calculated. A colorimeter was used to determine the chromaticity value of wild and cultivated GF， and the paraffin sections of them were prepared by safranin-fast green staining and examined under an optical microscope to observe their microstructure. Subsequently， the contents of water-soluble and alcohol-soluble extracts of wild and cultivated GF were detected by hot immersion method under the general rule 2201 in volume Ⅳ of the 2020 edition of the Pharmacopoeia of the People's Republic of China， the starch content was measured by anthrone colorimetric method， the content of total polysaccharides was determined by phenol-sulfuric acid colorimetric method， the sucrose content was determined by high performance liquid chromatography coupled with evaporative light scattering detection（HPLC-ELSD）， and the contents of representative components in them were measured by ultra-performance liquid chromatography（UPLC）. Finally， correlation analysis was conducted between quality traits and phenotypic traits， combined with multivariate statistical analysis methods such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）， key differential components between wild and cultivated GF were screened. ResultsIn terms of traits， the wild GF fruits were smaller， exhibiting reddish yellow or brownish red hues with significant variation between batches. While the cultivated GF fruits are larger， displaying deeper orange-red or brownish red. The diameter and individual fruit weight of cultivated GF were significantly greater than those of wild GF， while the blue-yellow value（b^*） of wild GF was significantly higher than that of cultivated GF. In the microstructure， the mesocarp of wild GF contained numerous scattered calcium oxalate cluster crystals， while the endocarp contained stone cell class round， polygonal or tangential prolongation， undeveloped seeds were visible within the fruit. In contrast， the mesocarp of cultivated GF contained few calcium oxalate cluster crystals， or some batches exhibited extremely numerous cluster crystals. The stone cells in the endocarp were predominantly round-like， with the innermost layer arranged in a grid pattern. Seeds were basically mature， and only a few immature seeds existed in some batches. Regarding primary metabolite content， wild GF exhibited significantly higher total polysaccharide level than cultivated GF（P<0.01）. In category-specific component content， wild GF exhibited significantly higher levels of total flavonoids and total polyphenols compared to cultivated GF（P<0.01）. Analysis of 12 secondary metabolites revealed that wild GF exhibited significantly higher levels of Shanzhiside， deacetyl asperulosidic acid methyl ester， gardenoside and chlorogenic acid compared to cultivated GF（P<0.01）. Conversely， the contents of genipin 1-gentiobioside， geniposide and genipin were significantly lower in wild GF（P<0.01）. ConclusionThere are significant differences between wild and cultivated GF in terms of traits， microstructure， and contents of primary and secondary metabolites. At present， the quality evaluation system of cultivated GF remains incomplete， and this study provides a reference for guiding the production of high-quality GF medicinal materials.

This article systematically analyzed the historical evolution of the origin， scientific name， producing area， quality evaluation， harvesting and processing， and other aspects of Quisqualis Fructus by consulting the ancient materia medica， medical books， prescription books， local literature and combining with the modern literature and standards， summarized and explored the development rules of its medicinal properties and efficacy along with their underlying causes， in order to provide support for the development and utilization of famous classical formulas containing this herb. According to the textual research， Shijunzi was first recorded as Liuqiuzi in Nanfang Caomuzhuang of the Jin dynasty， and the name of Shijunzi was first used in Kaibao Bencao of the Song dynasty， which has been consistently used throughout subsequent dynasties， and there were also aliases such as Junziren， Sijunzi， and Dujilizi. The mainstream source of Quisqualis Fructus used in the past dynasties has been the dried mature fruits of Quisqualis indica， a plant belonging to the family Combretaceae. In modern times， its variety Q. indica var. villosa has also been recorded as the medicinal material of Quisqualis Fructus. In 2007， the Flora of China（English edition） designated Q. indica var. villosa as a synonym of Q. indica. Today， the accepted name of Shijunzi is updated to Combretum indicum. According to ancient herbal records， the producing areas of Quisqualis Fructus were Guangdong， Hong Kong， Macao， Guangxi， Hainan， Sichuan and Fujian， and then gradually expanded to Yunnan， Taiwan， Jiangxi and Guizhou. Since the Song dynasty， two major production regions have gradually emerged in Sichuan， Chongqing and Fujian. Currently， it is primarily cultivated in Chongqing， Guangxi and other areas， with Chongqing yielding the highest output. Since modern times， superior quality has been defined by large size， a purple-black surface， plump grains， and a yellowish-white kernel. According to ancient herbal records， the harvesting period of Quisqualis Fructus was the July and August of the lunar calendar， mostly used raw after shelling or with the shell intact， it underwent processing methods such as cleaning， slicing， mixing， steaming， roasting， stewing， and frying. Currently， the harvesting period is autumn， followed by sun-drying or low-heat drying， with processing methods including cleaning， stir-frying， and stewing. In ancient and modern literature， the records of the properties， functions and indications of Quisqualis Fructus are basically the same， that is， sweet in taste， warm in nature， predominantly non-toxic， belonging to the spleen and stomach meridians. It possesses effects of insecticide， decontamination and invigorating spleen for ascariasis， enterobiasis， abdominal pain due to worm accumulation and infantile malnutrition.The contraindications for use primarily include avoiding consumption by individuals without parasitic infestations， limiting use for those with spleen-stomach deficiency-cold， refraining from drinking hot tea during medication， and avoiding excessive intake. Based on the textual research， it is suggested that the dried mature fruits of Q. indica should be used as the medicinal material for the development of famous classical formulas containing Quisqualis Fructus. Processing methods may be chosen according to prescription requirements， and the raw products is recommended for medicinal use if not specified.

ObjectiveThis study aims to explore the effect of Feiyanning granules on ferroptosis in lung cancer cells and its regulatory function within the nuclear transcription factor E₂-related factor 2 （Nrf2）/mouse solute carrier family 7 member 11 （SLC7A11）/glutathione peroxidase 4 （GPX4） signaling pathway. MethodsThe cell counting kit-8 （CCK-8） method was used to detect the effect of Feiyanning granule on the proliferation of A549 lung cancer cells. A549 lung cancer cells were categorized into a blank group， a ferroptosis inhibitor-1 （Fer-1） group （10 μmol·L^-1）， a Feiyanning granules （600 mg·L^-1） group， and a Feiyanning granules + Fer-1 group. After 48 hours of intervention， the activity and morphology of the cells were observed. The CCK-8 method was employed to measure cell viability. Biochemical assays were carried out to measure the levels of reactive oxygen species （ROS）， malondialdehyde （MDA）， glutathione （GSH）， and ferrous ions （Fe²⁺） in A549 cells. Western blot was utilized to evaluate the expression levels of Kelch-like ECH-associated protein 1 （Keap1）， Nrf2， SLC7A11， and GPX4 proteins. A549 lung cancer cells were categorized into a blank group and a Feiyanning Granule group （600 mg·L^-1）， and mitochondrial morphology was examined via transmission electron microscopy （TEM）. ResultsAfter the intervention of Feiyaning granules， the proliferation of A549 cells was significantly inhibited in a concentration-dependent manner compared with that in the blank group （P<0.01）. Compared with the blank group， the Feiyanning granules group exerted an significantly inhibitory effect on the viability of lung cancer cells （P<0.01）. Compared with that in the Feiyanning granules group， the cell viability in the Feiyanning granules +Fer-1 group was obviously restored （P<0.05）. Compared with the blank group， the Feiyanning Granule group showed a significant increase in the levels of ROS， MDA， and Fe²⁺ （P<0.01）， a significant decrease in the GSH level （P<0.01）， and facilitated ferroptosis. Compared with the blank group， the Feiyanning granules group showed significantly decreased expression of Nrf2， SLC7A11， and GPX4 proteins and enhanced expression of Keap1 （P<0.01）. Compared with those in the Feiyanning Granule group， the protein levels of Nrf2， SLC7A11， and GPX4 increased significantly （P<0.01）， and the expression of Keap1 decreased significantly in the Feiyanning granules + Fer-1 group （P<0.01）. Compared with the blank group， the Feiyaning granules group exhibited reduced mitochondrial size and increased matrix electron density. ConclusionFeiyanning granules can induce ferroptosis in lung cancer cells， and its underlying mechanism might be associated with the inhibition of the Nrf2/SLC7A11/GPX4 signaling pathway.

[摘 要] 目的：以MG-63球形细胞作为骨肉瘤干细胞（OSC）富集模型，探讨靶向细胞膜表面HSP90β的单抗2G1对其生物学特征的影响及其与Wnt信号相关蛋白变化的关系。方法：采用无血清悬浮培养人骨肉瘤MG-63细胞获得MG-63球形细胞。通过免疫荧光共定位和免疫共沉淀实验鉴定单克隆抗体2G1识别的靶抗原；采用流式细胞术检测MG-63亲本细胞和MG-63球形细胞膜表面HSP90β的表达水平；通过甲基纤维素成球实验、Transwell迁移和侵袭实验评价单抗2G1对MG-63球形细胞成球、迁移和侵袭能力的影响；采用裸鼠皮下移植瘤模型观察单抗2G1对MG-63球形细胞体内成瘤能力的影响；进一步结合转录组测序、京都基因与基因组百科全书（KEGG）通路富集分析及WB检测，分析单抗2G1处理后相关分子通路及干细胞特性、上皮-间充质转化（EMT）、Wnt/β-catenin信号通路相关蛋白表达变化。结果：免疫荧光共定位和免疫共沉淀结果显示，单抗2G1识别的靶抗原为HSP90β。流式细胞术结果显示，MG-63球形细胞膜表面HSP90β阳性表达细胞比例为（20.77 ± 1.29）%，显著高于MG-63亲本细胞的（2.87 ± 0.33）%（P < 0.001）。与对照组相比，单抗2G1处理后MG-63球形细胞成球数由82.67 ± 4.70降至42.67 ± 4.00，迁移细胞数由117.00 ± 5.57降至78.33 ± 5.03，侵袭细胞数由130.70 ± 5.03降至89.33 ± 5.03（均P < 0.001）。裸鼠移植瘤实验显示，单抗2G1组移植瘤生长受到抑制，瘤质量抑制率和瘤体积抑制率分别为（31.83 ± 16.36）%和（22.00 ± 15.48）%。转录组测序共筛选出698个差异表达基因，其中329个基因上调，369个基因下调；KEGG富集分析显示差异表达基因涉及Wnt信号通路。WB法检测结果显示，与对照组相比，单抗2G1组SOX2、OCT4、N-cadherin、p-β-catenin和c-Myc表达下调，E-cadherin表达上调（均P < 0.001）。结论：细胞膜表面HSP90β在MG-63球形细胞中高表达，靶向HSP90β的单抗2G1可抑制MG-63球形细胞的成球、迁移、侵袭及体内成瘤能力，并影响其干细胞标志物、EMT相关标志物及Wnt/β-catenin信号通路蛋白的表达水平，提示HSP90β可能参与调控MG-63球形细胞的OSC样特征，可作为骨肉瘤靶向治疗的潜在研究靶点。

ObjectiveTo investigate the protective effect and mechanism of action of flavonoid combination of Alpiniae Officinarum Rhizoma （A. officinarum） against Helicobacter pylori （H. pylori） gastritis in mice. MethodsAfter acclimatization for one week， 56 SPF-grade healthy C57BL/6J mice were gavaged with mixed antibiotics for three consecutive days. They were randomly divided into a normal group， model group， positive drug group （triple therapy group）， and low- and high-dose groups （100， 200 mg·kg^-1） of flavonoid combination of A. officinarum. The H. pylori gastritis mice model was established by gavage with H. pylori bacterial suspension in each group except for the normal group. After successful modeling， mice were administrated with corresponding drugs once a day for two weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in gastric tissue. Rapid urease test paper was used to detect the positive rate of H. pylori. Silver staining was used to observe the H. pylori adherence on the surface of gastric tissue. Immunohistochemistry was used to detect the protein expression of interleukin-8 （IL）-8 and myeloid differentiation factor （MyD88） in gastric tissue. The serum levels of IL-6， tumor necrosis factor-α （TNF-α）， IL-8， and IL-1β were detected by enzyme-linked immunosorbent assay （ELISA）. The expressions of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） protein were detected by Western blot. ResultsCompared with those in the normal group， mice in the model group had lower gastric weight coefficients， higher pH of gastric juice， 100% H. pylori infection rate， and significantly changed gastric histopathology. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the model group were significantly elevated， and the serum levels of inflammatory factors IL-6， TNF-α， IL-8， and IL-1β were significantly up-regulated in mice. Compared with that in the model group， the gastric weight coefficient of mice in each treatment group of the flavonoid combinations of A. officinarum was elevated （P<0.01）， and the pH of gastric juice was reduced （P<0.01）. The infection rate of H. pylori was reduced. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the treatment groups were significantly reduced （P<0.01）， and the serum levels of inflammatory factors IL-6， TNF-α， IL-8， and IL-1β were significantly reduced in a dose-dependent manner （P<0.01）. The flavonoid combinations of A. officinarum down-regulated the expression of PI3K and Akt proteins in H. pylori gastritis-infected cells （P<0.01）. ConclusionThe protective effect of flavonoid combinations of A. officinarum against H. pylori gastritis is associated with the inhibition of H. pylori infection rate and regulation of PI3K/Akt signaling pathway， resulting in inhibiting the release of inflammatory factors.

Objective:This study aimed to investigate the regulatory role and mechanism of myosin heavy chain 9 (MYH9) in the invasion of Zika virus (ZIKV) into human glioma cells (U251).Methods:Utilizing CRISPR/Cas9 technology, MYH9-knockout U251 cells (U251-MYH9 KD) were constructed. Following ZIKV infection, the protein expression levels, RNA load, and viral titer of ZIKV were detected through western blot (WB), Real-time fluorescence quantitative polymerase chain reaction (qPCR), and plaque formation assays, respectively. The infection efficiency of ZIKV in U251 cells treated with the MYH9 inhibitor blebbistatin was assessed. The binding and internalization efficiency of ZIKV were measured in U251-MYH9 KD cells. The interaction between MYH9 and the ZIKV envelope protein (E) was studied using co-immunoprecipitation (Co-IP). The effects of soluble MYH9 recombinant protein and anti-human MYH9 antibodies on ZIKV infection were evaluated by qPCR and plaque formation assays. Results:It was found that knockout or inhibition of MYH9 significantly suppressed ZIKV infection in U251 cells. MYH9 knockout notably inhibited the binding and internalization of ZIKV in U251 cells. MYH9 interacted with the ZIKV E protein, and both MYH9 recombinant protein and anti-human MYH9 antibodies, by blocking the binding of ZIKV E protein to cell surface MYH9, inhibited ZIKV infection in U251 cells in a dose-dependent manner.Conclusions:MYH9 facilitates ZIKV invasion into U251 cells through interaction with the ZIKV E protein.

By sorting out the records of Atractylodis Macrocephalae Rhizoma as food and medicine, this article believed that Atractylodis Macrocephalae Rhizoma as the same origin of medicine and food for a long history, the earliest in the Qin and Han Dynasty Shen Nong Ben Cao Jing: "Zhu, bitter and warm, having efficacy for cold, dampness, paralysis of dead muscles, and spasm; antiperspirant, removing heat, digesting food, and be used as decoction bait; taking for a long time, losing weight and prolonging life, and don't be hungry". On the basis of Shen Nong Ben Cao Jing, later doctors continue to conduct long-term clinical observation and compatibility of its medicine and food efficacy, which is widely used in internal, external, gynaecology, pediatrics, skin and other departments, to treat spleen and stomach diseases, epidemic diseases, kidney diseases, etc. At the same time of clinical practice, Atractylodis Macrocephalae Rhizoma is also widely used in the folk for dietary therapy, including decoction bait, ointment, decoction, soup and other edible methods. Therefore, as a medicine and food and health preservation product, Atractylodis Macrocephalae Rhizoma has great development potential in daily health care and disease diagnosis and treatment with the effects of non-toxic, satiety and longevity.

Objective To investigate the risk factors for pneumonia complicating infectious mononucleosis(IM)in adults and construct a nomogram prediction model.Methods A retrospective analysis was conducted on 198 IM patients admitted to the Second Affiliated Hospital of Air Force Medical University from January 2015 to December 2021.Patients were divided into pneumonia group(n=52)and non-pneumonia group(n=146)based on whether pulmonary infection occurred during hospitalization.The baseline data(age,gender,place of onset,etc.),clinical manifestations(maximum body temperature,lymph node enlargement,splenomegaly,etc.),and inflammatory indicators[white blood cell count(WBC),C-reactive protein(CRP),etc.]were compared between the two groups.Kaplan-Meier curves were plotted to analyze the key indicators affecting the hospital stay of IM patients.Multivariate logistic regression was used to analyze the independent risk factors for pneumonia complicating IM in adults and construct a nomogram prediction model based on the identified risk factors.The predictive efficacy of the model was evaluated using the receiver operating characteristic(ROC)curve and the consistency of the model was assessed using the calibration curve.The fit of the model was evaluated using the Hosmer-Lemeshow test.Additionally,the sensitivity,specificity,and accuracy of the model were assessed using confusion matrix.Results Compared with non-pneumonia group,the pneumonia group had a significantly higher proportion of patients from rural areas,with body mass index(BMI)≥24 kg/m2,smoking history,hepatomegaly,fever duration of≥7 d,as well as increased total hospitalization costs and average daily hospitalization costs,and prolonged hospital stay(P<0.05).The proportion of patients with a history of antibiotic use was lower in the pneumonia group(P<0.05).Kaplan-Meier survival analysis showed that patients from rural areas,with BMI≥24 kg/m2,smoking history,no prophylactic use of antibiotics,fever duration≥7 d,and hepatomegaly had significantly prolonged hospital stays(P<0.05).Multivariate logistic regression analysis revealed that living in a rural area(OR=4.089,P<0.05),hepatomegaly(OR=4.082,P<0.05),and elevated WBC(OR=1.205,P<0.05)were independent risk factors for pneumonia complicating IM in adults,while the prophylactic use of antibiotics(OR=0.142,P<0.05)was an independent protective factor.The area under the ROC curve of the constructed nomogram prediction model was 0.827(95%CI 0.762-0.892),and the slope of the calibration curve was close to 1,and the Hosmer-Lemeshow test showed χ2=5.299,P=0.725,indicating good consistency and fit of the prediction model.The results of the confusion matrix assessment showed that the sensitivity of the model was 0.669(0.624-0.773),the specificity was 0.827(0.724-0.930),and the accuracy was 0.732(0.665-0.793).Conclusion The nomogram prediction model based on place of onset,hepatomegaly,the prophylactic use of antibiotics and WBC has excellent fit and discrimination,providing an effective quantitative tool for prognosis assessment of IM.

Objective To investigate the association of digit ratio with single nucleotide polymorphism(SNP)at three loci(rs17576,rs3918249,rs9509)of matrix metallopeptidase 9(MMP-9)gene.Methods A total of 804 Ningxia Han youths(399 males and 405 females)were used as the study subjects.A digital camera was used to take frontal photographs of the hands,and image analysis software was used to mark the anatomical points and measure the lengths of each finger of both hands(2D,3D,4D,5D);Multiplexed PCR was used to detect the three polymorphic sites of the MMP-9 gene,SPSS 25.0 and R Studio software were used for data analysis and plotting.Results The 2D/3D(P＜0.05)and 2D/4D(left,P＜0.01,right,P＜0.05)of both hands,2D/5D(P＜0.01),3D/5D,4D/5D(P＜0.05)of the right hand,and 3D/4D(P＜0.05)of the left hand in female youths of Ningxia Han were significantly higher than those in males,Differences in genotypes and allele frequencies at all 3 loci of the MMP-9 gene were not statistically significant between genders(P＞0.05).Right hand 2D/4D was significantly associated with genotypes at the rs17576 and rs3918249 loci in male youths(P＜0.05).Conclusion MMP-9 gene SNPs(rs17576 and rs3918249)may be associated with the formation of 2D/4D of Ningxia Han male youths.