ObjectiveTo characterize the quality differences among different germplasm and introduced varieties of Bupleurum scorzonerifolium roots（BSR）， and explore the underlying molecular mechanisms， providing a basis for high-quality production and quality control. MethodsWild BSR from Yulin（YLW） served as the quality reference， we conducted comparative analysis among YLW， locally domesticated wild germplasm in Yulin（YLC3）， Daqing germplasm introduced and cultivated in Yulin（YLDQC3）， and locally cultivated germplasm in Daqing（DQC3）. A combination of traditional pharmacognostic methods and modern multi-omics analyses was employed， including macroscopic traits（appearance， odor）， microscopic features（proportions of cork， phloem， xylem）， cell wall component contents（hemicellulose， cellulose， lignin）， carbohydrate contents（starch， water-soluble polysaccharides）， marker compound contents（ethanol-soluble extracts， total saponins， liposoluble extracts， and saikosaponins A， B₂， C， D）， metabolomics， and transcriptomics， in order to systematically characterize quality differences and investigate molecular mechanisms among these samples. ResultsMacroscopically， Yulin-produced BSR（YLW， YLC3， YLDQC3） exhibited significantly greater weight， length， and upper and middle diameters than Daqing-produced BSR（DQC3）. Odor-wise， YLW and YLC3 had a a fragrance taste， YLDQC3 had a rancid oil odor， and DQC3 had a sweet and fragrant taste. Microscopically， Yulin germplasm（YLW， YLC3） and Daqing germplasm（YLDQC3， DQC3） shared similar structural features， respectively. However， Yulin germplasm showed significantly higher proportions of cork and phloem， as well as stronger xylem vessel staining intensity compared to Daqing germplasm. Regarding various component contents， Yulin germplasm contained significantly higher levels of ethanol-soluble extracts， total saponins， and saikosaponins A， B₂， C， D， while Daqing germplasm had significantly higher levels of hemicellulose， starch， and liposoluble extracts. After introduction to Yulin， the Daqing germplasm（YLDQC3） showed increased starch， water-soluble polysaccharides and liposoluble extracts contents， decreased cell wall component content， but no significant difference in other component contents. Metabolomics revealed that saponins and terpenes accumulated significantly in Yulin germplasm， while alcohols and aldehydes accumulated predominantly in Daqing germplasm. Transcriptomics indicated similar gene expression patterns within the same germplasm but specificity between different germplasms. Integrative metabolomic-transcriptomic analysis identified 145 potential key genes associated with the saikosaponin biosynthesis pathway， including one acetyl-coenzyme A（CoA） acetyltransferase gene（ACAT）， one 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene（HMGS）， two hydroxymethylglutaryl-CoA（HMG-CoA） reductase genes（HMG）， one phosphomevalonate kinase gene（PMK）， one 1-deoxy-D-xylose-5-phosphate synthase gene（CLA）， one hydroxymethylbuten-1-aldol synthase gene（HDR）， two farnesyl pyrophosphate synthase genes（FPPS）， one squalene synthase gene（SQS）， one β-amyrin synthase gene（BAS）， 102 cytochrome P450（CYP450） gene family members， and 32 uridine diphosphate-glucuronosyltransferase（UGT） gene family members. ConclusionAmong the three cultivated types， YLC3 most closely resembles YLW in appearance， microscopic features， contents of major bioactive constituents， metabolomic and transcriptomic profiles. Yulin germplasm exhibits superior saponin synthesis capability compared to Daqing germplasm， and Yulin region is more suitable for the growth of B. scorzonerifolium. Based on these findings， it is recommended that artificial cultivation in northern Shaanxi and similar regions utilize the local Yulin germplasm source cultivated for at least three years.

ObjectiveBased on the traditional quality evaluation methods summarized in previous dynasties， this paper systematically contrasted the quality differences between wild Polygalae Radix（WPR） and cultivated Polygalae Radix（CPR） from the aspects of character， microscope and chemical composition by modern scientific and technological means， providing a basis for high-quality production and quality control. MethodsCPR and local WPR in Yulin city， Shaanxi province from 1 to 6 years were collected， and a systematic comparative analysis was conducted using traditional pharmacognosy research methods combined with modern multi-omics analysis techniques， including character traits（length， weight， diameter）， cross-sectional microscopic features（proportions of cork， phloem， xylem， etc）， cell wall component content（hemicellulose， cellulose， lignin）， extracts content（water-soluble extract and alcohol-soluble extract）， carbohydrate content（starch， water-soluble polysaccharides）， contents of total flavonoids， total saponins and specific marker compounds（3，6′-disinapoyl sucrose， polygalaxanthone Ⅲ， tenuifoliside A， tenuifoliside C， sibiricose A₅ and A₆） and other indexes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was employed to conduct comparative analysis of secondary metabolites in WPR and CPR， and multivariate statistical analysis such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were combined to screen the key differential components of them. ResultsIn terms of appearance， there were significant differences between WPR and CPR. The characteristics of WPR conformed to the "thick wrinkles on the epidermis" recorded in ancient books， featuring a wrinkled surface and grayish-brown appearance. However， CPR had a finer texture and a yellowish white appearance， with weight， length， and diameter increasing with longer cultivation periods. In terms of microscopy， WPR exhibited a thick cork layer with fissures in the phloem， whereas CPR had a thinner cork layer with uniformly arranged cork cells. Younger PR specimens showed numerous phloem fissures in cross-sections， while older specimens display progressively denser arrangements of phloem parenchyma cells. In terms of the contents of various major components， the contents of water-soluble extract， starch and total saponins in WPR were inversely proportional to the root diameter， while the contents of water-soluble extract， water-soluble polysaccharides and total saponins in CPR decreased with the increase of planting years. The content of xanthones in WPR was significantly higher than that of CPR， while the contents of other major components showed no significant change pattern. Among the six indicator components， the average content of sibiricose A₅ in WPR was significantly higher than that of CPR， followed by slightly higher content of tenuifoliside A. In CPR， the relative content of 3，6′-disinapoyl sucrose and tenuifoliside A was the highest. The former showed an increase in volatility with increasing cultivation years， while the latter showed a decrease in volatility. The results of differential compound analysis based on UPLC-Q-TOF-MS showed that there were significant differences in metabolites between WPR and CPR samples. Among them， the seven compounds with the largest differences among WPR samples of different thicknesses were polygalasaponins， and for CPR with different planting years， the main differential compounds were oligosaccharide esters. ConclusionThere are differences between WPR and CPR in character， microscopic structure and chemical composition， and some components are inversely proportional with the increase of diameter and cultivation duration due to the distribution characteristics. However， the longer the cultivation years of PR， the closer it is to the "thick wrinkles on the epidermis" of WPR， which has been respected by generations. It is suggested that this traditional character combined with modern component contents should be used as the index of artificial cultivation and quality control of PR.

Microglial functions are linked to Ca²⁺ signaling, with endoplasmic reticulum (ER) calcium stores playing a crucial role. Microglial abnormality is a hallmark of Alzheimer's disease (AD), but how ER Ca²⁺ receptors regulate microglial functions under physiological and AD conditions remains unclear. We found reduced ryanodine receptor 2 (Ryr2) expression in microglia from an AD mouse model. Modulation of RyR2 using S107, a RyR-Calstabin stabilizer, blunted spontaneous Ca²⁺ transients in controls and normalized Ca²⁺ transients in AD mice. S107 enhanced ATP-induced migration and phagocytosis while reducing ramification in control microglia; however, these effects were absent in AD microglia. Our findings indicate that RyR2 stabilization promotes an activation state shift in control microglia, a mechanism impaired in AD. These results highlight the role of ER Ca²⁺ receptors in both homeostatic and AD microglia, providing insights into microglial Ca²⁺ malfunctions in AD.
Animals ; Microglia/pathology* ; Alzheimer Disease/pathology* ; Phagocytosis/drug effects* ; Ryanodine Receptor Calcium Release Channel/metabolism* ; Disease Models, Animal ; Mice ; Cell Movement/drug effects* ; Mice, Transgenic ; Calcium Signaling/physiology* ; Calcium/metabolism* ; Mice, Inbred C57BL ; Male ; Endoplasmic Reticulum/metabolism*

Karacoline is a compound found in the plant Aconitum kusnezoffii Reichb. Although Aconitum kusnezoffii Reichb is widely used for the treatment of pain, very few studies have been carried out on the use of karacoline due to its potential toxicity. In this study, we selected key matrix metalloproteinases (MMPs), collagen II, and aggrecan as targets due to their association with intervertebral disc degeneration (IDD). Using these targets, we then used network pharmacology to predict a series of molecules that might exert therapeutic effects on IDD. Of these molecules, karacoline was predicted to have the best effect. Tumor necrosis factor (TNF)-αis known to promote the degeneration of the extracellular matrix in IDD. We therefore applied different concentrations of karacoline (0, 1.25, or 12.88μM) along with 100 ng/mL TNF-αto rat nucleus pulposus cells and found that karacoline reduced the expression of MMP-14 in IDD by inhibiting the nuclear factor (NF)-κB pathway, while collagen II and aggrecan expression was increased. This suggested that extracellular matrix degradation was inhibited by karacoline (P<0.05). Our data therefore reveal a new clinical application of karacoline and provide support for the use of network pharmacology in predicting novel drugs.

Objective To evaluate the differences of serum RANTES(regulated on activation, normal T cell expressed and secreted), MCP-1 (monocyte chemoattractant protein), and SDF-1β (stromal cell-derived factor-1β) in patients with acquired immunodeficiency syndrome (AIDS) and healthy people.Methods 38 AIDS patients who were admitted to a hospital between January 2010 and January 2015 were as AIDS groups, 38 healthy persons were as a healthy group, serum levels of RANTES, MCP-1, and SDF-1β in two groups were detected, and the subgroup analysis was carried out according to the viral load.Results Serum levels of RANTES, MCP-1, and SDF-1β in AIDS group were (1 392.55±227.69)pg/mL,(450.91±103.04)pg/mL, and(104.82±22.52)pg/mL respectively,all were significantly higher than those in healthy group([120.58±55.87] pg/mL, [74.25±33.62] pg/mL, and [39.04±11.43]pg/mL respectively)(all P<0.05).Among AIDS patients with HIV viral load 4≤Log(VL)<5 and Log(VL)≥5, serum RANTES were (1 470.34±155.01)pg/mL and (1 408.29±181.54)pg/mL respectively,which were both significantly higher than patients with HIV viral load Log(VL)<4([1 183.12±174.54]pg/mL);serum MCP-1 and SDF-1β levels in AIDS patients with HIV viral load 4≤Log (VL)<5 were (537.93±89.32)and(149.31±18.05)pg/mL respectively,which were significantly higher than patients with HIV viral load Log(VL)≥5([410.26±80.57] pg/mL, [81.53±20.31]pg/mL respectively) and HIV viral load Log(VL)<4([381.71±77.26] pg/mL, [72.90±21.62]pg/mL respectively), differences were both statistically significant(both P<0.05).Conclusion Serum levels of RANTES, MCP-1, and SDF-1β are significantly increased in AIDS patients, which are related to the level of viral load.

Objective To explore the expression of CD44v8 protein in human bladder and urothelial carcinoma ,as well as the value in diagnosis of human bladder and urothelial carcinoma .Methods RT‐PCR was used to analysis the expression of CD44v8 protein in 75 patients with bladder and urothelial carcinoma in the pathological stage and clinical stage and 20 subjects of normal bladder mucosa were collected as control .Results CD44v8 protein was negative in all normal bladder and urothelial mucosa ,the copy number was less than 1 × 102 copy/mL ;26 cases of bladder and urothelial carcinoma was positive ,and the positive rate of CD44v8 was 34 .7% ,and Ct values were less than 35 and copy number was greater than 1 × 104 copy/mL .Positive rate was correla‐ted with high pathological grades and TNM stages ,but no significant difference was observed in recurrence of tumor .Conclusion CD44v8 could be useful indicator for the assessment of pathological grades and TNM stages of bladder and urothelial carcinoma .

BACKGROUND:Dorsal digital block refers to the commonly used anesthesia for adults in smal or moderate hand injury surgeries, but in recent years, modified transthecal digital block technique is gradualy respected, which is favored with a rapid and good effect and fewer complications.
OBJECTIVE:To evaluate the clinical anesthetic outcomes of modified transthecal digital block and traditional dorsal digital block technique for the treatment of hand injury of adults in emergency by a prospective randomized controled study.
METHODS:Totaly 60 adult patients with hand injury were enroled and divided into two groups of modified transthecal digital block and traditional dorsal digital block randomly. Blocks were performed by one single surgeon. The operation time, local anesthetic dose, onset time of anesthesia, duration of anesthesia, success rate of anesthesia, visual analogue scale scores and complications were recorded.
RESULTS AND CONCLUSION:The anesthesia effects in the two groups were acceptable. There was no significant difference in the onset time of anesthesia, duration of anesthesia, success rate of anesthesia and complications between the two groups (P > 0.05). The operation time of anesthesia, local anesthetic dose, and visual analogue scale scores were significantly different between the two groups (P< 0.05). Modified transthecal digital block is more convenient and has less pain than the traditional root digital block, which is a safe and reliable anesthetic technique.

Objective To explore CT manifestion of multiple primary lung cancer (MPLC)and to improve the recognization of MPLC.Methods The CT manifestions of 12 cases with MPLC proved by pathology were retrospectively reviewed.Results All the 12 cases were double primary lung cancer.There were 24 lesions with 21 peripheral and 3 central,the average diameter was (2.2 ± 0.6)cm.Lesions located in contralateral lobes were in 2 patients,and located in ipsilateral lobes were in 10 patients,with 3 located in the same lobe and 7 in the different lobes.There were 7 cases of adenocarcinoma-adenocarcinoma,3 cases of squamous cell carci-noma-adenocarcinoma,1 case of squamous cell carcinoma-squamous cell carcinoma and 1 case of squamous cell carcinoma-carcinoid. 1 1 cases were metachronous and 1 case was synchronous.3 lesions were lump located at hilus of the lung,21 lesions were intrapul-monary nodules,showing masses with lobulated shape,spicules of margin,vascular convergence,vacuole and pleural indentation sign.Most foci displayed moderate intensity enhancement and homogeneous density in triphase enhanced scans,the CT value of le-sions on enhanced images ranged from 20-60 HU.Conclusion MPLC are synchronous and peripheral adenocarcinoma type,all of the lesions have typical CT features of primary lung cancer.

We obtained recombinant ferritin from Dermatophagoides f arinae ,and analyzed the characterization of the pro‐tein .A pair of primers was designed according to the known sequence of ferritin gene .The live mites identified and cultured lo‐cally were picked and the total RNA was extracted .The ferritin gene fragment was amplified by RT‐PCR ,and cloned into pET32a vector ,and then transferred into E .coli Top10 .The target gene obtained from the recombinant plasmid by digestion with Bam HⅠand Hind Ⅲ was connected to the prokaryotic expression vector pET‐32a .The expressed recombinant plasmid containing ferritin gene was constructed by cloning target gene into pET‐32a and transferred into E .coli Bl21 (DE3) .The ex‐pressed recombinant protein was analyzed by SDS‐PAGE ,and was purified by immobilized metal ion affinity chromatography (IMAC) .The ferritin expressed by dust mite was analyzed by the method of bioinformatics .The recombinant plasmid pET32a‐ferritin was constructed .SDS‐PAGE showed a correct molecular weight of the recombinant ferritin protein .After purification by affinity chromatography ,the protein showed only one strip on SDS‐PAGE gel .SDS‐PAGE showed a band at 20 kD .Dust mite ferritin contains 8 serine kinase ,7 threonine kinase ,7 tyrosine kinase ,and 0 histidine kinase phosphorylation sites .Hy‐drophilic region is larger than the hydrophobic region and it is an unstable protein .In conclusion ,the ferritin gene has been cloned and expressed .The purified ferritin has high purity . The study provides a basis for further study of composition and physicochemical properties of house dust mite allergen .

BACKGROUND:Both in vitro and in vivo studies have confirmed that, bone morphogenetic protein (BMP) regulates the differentiation of osteoblasts and chondroblasts, induces heterotopic bone formation, promotes fracture healing, and controls the morphology of skeleton in mammals.
OBJECTIVE:To treat chronic bone defects using particle gun containing BMP2 gene eukaryotic expression plasmid via local injection.
METHODS:A total of 72 healthy New Zealand white rabbits were applied to establish chronic bone defect model in the rabbit radius. According to the length of bone defect, the rabbits were divided into three groups:1.5 cm group, 2.0 cm group, 2.5 cm group. Each group was further randomly assigned into two subgroups:treatment group (BMP-2 gene transfection) and control group (natural y healing). X-ray examinations were performed at 1, 3, 8 and 9 weeks after transfection, and soft tissue between the bone defects was harvested to detect BMP-2 using western blot analysis;and radius specimens were taken for gross observation at the same time points, to evaluate the healing.
RESULTS AND CONCLUSION:(1) Gross specimen observation:bone cal us formation in treatment group was general y more than that in control group. (2) Lane-Sandhu X-ray score in treatment group was significantly higher than that in control group at 1, 3, 8, 9 weeks after transfection (P<0.05). (3) BMP-2 concentration in treatment group was significantly higher than that in control group at each time point (P<0.05). The local transfer of particle gun-mediated BMP-2 gene is an effective therapy of chronic bone defect.