The new generation of artificial intelligence technology,represented by deep learning,has emerged as a crucial driving force in the advancement of new drug research and development.This article creatively proposes a workflow named"Molecular Factory"for the design and optimization of drug molecules based on artificial intelligence technology.This workflow integrates intelligent molecular generation models,high-performance molecular docking algorithms,and accurate protein-ligand binding affinity prediction methods.It has been integrated as a core module into DrugFlow,a one-stop drug design software platform,providing a comprehensive set of mature solutions for the discovery and optimization of lead compounds.Utilizing the"Molecular Factory"module,we conducted the research of second-generation inhibitors against Menin that can combat drug resistance.Through the integration of computational and experimental approaches,we rapidly identified multiple promising compounds.Among them,compound RG-10 exhibited the IC50 values of 9.681 nmol/L,233.2 nmol/L,and 40.09 nmol/L against the wild-type Menin,M327I mutant,and T349M mutant,respectively.Compared to the positive reference molecule SNDX-5613,which has entered Phase Ⅱ clinical trials,RG-10 demonstrated significantly enhanced inhibitory activity against the M327I and T349M mutants.These findings fully demonstrate the unique advantages of the"Molecular Factory"technology in practical drug design and development scenarios.It can rapidly and efficiently generate high-quality active molecules targeting specific protein structures,holding significant value and profound implications for advancing new drug discovery.

OBJECTIVE：To improve the quality standard of M ongolian med icine Artemisia sacrorum ，and to provide scientific basis for comprehensive quality evaluation. METHODS ：The appearance and microscopic characteristics of A. sacrorum were identified；scopoletin，chlorogenic acid ，caffeic acid ，scopoletin and 3，5-dicaffeoylquinic acid were identified quantitatively by TLC；the contents of above 5 components were determined by HPLC. The water content ，total ash and extract were examined. RESULTS：The stem of A. sacrorum was cylindrical ，and its surface was purple or purple-brown or cyan-brown ；the leaves were ovate or oblong-ovate ，fragrant；the flowers were yellow ，head-shaped，subglobose or hemispherical. The powder was green or yellow-green，its pollen grain had three germination ；the parenchymal cell clusters with sharp edges and numerous threaded ducts ， occasionally having marginal pitted ducts ；its wood fibers were in bundles mostly. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for 5 substance control and samples. The linear range of scopoletin， chlorogenic acid ， caffeic acid ， scopolactone and 3，5-dicaffeoylquinic acid were 85.60-428.00， 10.16-101.60， 10.20-102.00，40.84-408.40 and 40.80-408.00 μg/mL（all r＞0.999 0）. RSDs of precision ，stability，repeatability tests were all less than 3.00％（n＝6）. The average recoveries were 103.07％，99.66％，98.37％，97.78％，98.40％（all RSDs ＜3.00％，n＝6）. The contents of the above-mentioned 5 compounds in 10 batches of samples were 0.36％-1.23％，0.09％-0.51％，0.04％-0.13％， 0.61％ -1.13％ ，0.12％ -1.11％ ，respectively；the average com contents of water ，total ash and water soluble extract were 6.25％，5.86％，26.50％，respectively. CONCLU SIONS：O the basis of the original quality standard of A. sacrorum ， microscopic identification，TLC identification ，content determination and examination items of water ，total ash and extract are added. The method shows good precision ，accuracy and stability ，which can provide reference for more scientific and standardized evaluation of the quality of this medicinal material.

Objective To revise the Symptom Distress Scale for postoperative patients with pituitary tumor and to test its reliability and validity. Methods On the base of previous qualitative interview and literature review, Delphi consultation was performed to identify items of the Symptom Distress Scale for postoperative patients with pituitary tumor. By convenience sampling method, totally 191 patients from four first-class ternary hospitals in Jiangsu province were investigated effectively by this scale. Results A scale of 4 factors and 16 items was identified by expert interviews, item analysis, exploratory factor analysis and the four factors could explain 69.812%of the variance. The Cronbachαcoefficient of the scale was 0.920, the content validity index was 0.915, and the interrater reliability was 0.860. Conclusions Symptom Distress Scale for postoperative patients with pituitary tumor has good reliability and validity to assess the symptom distress of pituitary tumor patients after operation.

Objective To investigate the effect and mechanism of human amniotic epithelial cells (hAECs) transplantation in treatment of cerebral hemorrhage in rabbits.Methods Thirty New Zealand white rabbits were used to induce cerebral hemorrhage.Animals were divided into hAECs group and isotonic saline group according to random number table,with 15 rabbits per group.Before transplanted to rabbits,hAECs were transfected with the retrovirus carrying green fluorescent protein (GFP).Morphologic and behavioral changes in both groups were noted periodically.Survival of transplanted hAECs and expressions of glial fibrillary acidic protein (GFAP) and microtubule-associated protein 2 (MAP-2)in focal cerebral tissues were observed.Results In hAECs group,the rabbits obtained progressive recovery in walking,supporting and coordinated motion.Restoration period mostly ranged from 2-3 weeks.Most of the rabbits in hAECs group had limb motor function recovered to grade Ⅱ-Ⅲ,while the recovery is slow in isotonic saline group with most ranging from grade Ⅰ-Ⅱ.According to Tarlov score,limb motor function presented significant difference between hAECs group and isotonic saline group (P ＜ 0.05).Immunohistochemical staining showed positive expressions of GFAP and MAP-2 in hAECs group,but no expressions in isotonic saline group (P ＜ 0.05).Conclusion hAECs transplantation effectively improves neural behavior and reduces nerve function impairment in treatment of cerebral hemorrhage in rabbits.

Objective To study the clinical significance of the injury and functional change of hypothalamic-pituitary-adrenal (HPA) axis after acute severe traumatic brain injury (TBI) in the rats.Methods A total of 60 adult healthy male Spraque-Dawley rats were randomly (random number) divided into 3 groups (n =20 in each group):sham operation group,model group and treatment group.The TBI models of rats were established by Feeney' s method.A low dose of dexamethasone (0.6 mg/kg) was injected into the abdominal cavity 20 minutes,24 hours and 48 hours after injury in treatment group,while rats of sham operation group and model group received equal volume of normal saline instead.All the rats were injected 1 μg adrenocorticotropic hormone (ACTH) into the abdominal cavity.The related parameters were detected at four time points,3 hours,12 hours,24 hours and 72 hours after cerebral contusion.The plasma corticosterone (CORT) and ACTH levels were measured by chemiluminescence.The hypothalamic,pituitary and adrenal of the rats were taken out for observing interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) expression detecting by immunohistochemical techniques at 72nd hour after TBI.One-way ANOVA and SNK-q test were used to analyze the results with SPSS 17.0 software package.Results The levels of ACTH and CORT on 3rd hour of model group raised remarkably compared with that of sham operation group,then they reduced gradually.The levels of CORT were lower than that of sham operation group at every time points after ACTH stimulation test (P ＜0.05 or P ＜0.01).The levels of CORT at all time points of treatment group were changed remarkably compared with that of model group.However,the ACTH levels of treatment group on 24 h increased slightly than that of model group.And the tendency of them was similar to model group (P ＜ 0.05 or P ＜ 0.01).The number of the hypothalamus and pituitary cells which express IL-6 and TNF-α in model group was more significantly increased when compared with that in sham operation group (P ＜ 0.01),while the number of this kind of cell in treatment group was significantly decreased than that in model group (P ＜ 0.01).The number of the adrenal cortex cells which express IL-6 in treatment group was more significantly decreased when compared with that in model group (P＜ 0.01),while the number of this kind of cell in model group was significantly increased than that in sham operation group (P ＜ 0.01).However,there was no significant difference of the TNF-α between all the groups (P ＞ 0.05).Conclusions Functional change of adrenal occurs early in the severe acute traumatic brain injury rats,and the response of adrenal to ACTH decreased as time goes by.Low-dose,short-course dexamethasone can delay the pathological changes,reduce the inflammatory response of HPA axis and increase the sensitivity of adrenal response to ACTH.

OBJECTIVEWe examined alterations in the expression of tumorigenesis-related genes in the pituitary gland of rats exposed to electromagnetic pulses (EMP).

METHODSThe global gene expression profiles of the pituitary gland in EMP-exposed and control groups were detected by cDNA microarray analysis. We then validated and further investigated the reduced expression of two tumorigenesis-related genes, Pten, and Jund, by assessing their mRNA and protein expression by quantitative real-time-PCR, western blotting, and immunohistochemistry in the pituitary gland of rats 6 months after exposure to EMP.

RESULTSEMP exposure induced genome-wide gene expression changes in the rat pituitary gland. There was decreased expression of the Pten and Jund mRNAs and proteins in EMP-exposed rats compared with in unexposed control animals.

CONCLUSIONEMP exposure alters the expression of tumorigenesis-related genes in the pituitary gland. These tumorigenesis-related genes are potentially involved in the development of pituitary gland tumors in rats.

Adenoma ; genetics ; metabolism ; pathology ; Animals ; Down-Regulation ; Electromagnetic Phenomena ; Female ; Gene Expression Profiling ; Oligonucleotide Array Sequence Analysis ; PTEN Phosphohydrolase ; genetics ; metabolism ; Pituitary Gland ; metabolism ; Pituitary Neoplasms ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; Rats ; Rats, Wistar ; Real-Time Polymerase Chain Reaction

Fresh roots of healthy four-year-old American ginseng were randomly divided into two groups and stored for 180 days in sand without washing (treatment A) and in plastic bag with preservative after washing (treatment B), and sampled after 45, 90, 135 and 180 days of storage, respectively. The incidence of disease was surveyed and the contents of ginsenoside, polysaccharide, and free amino acids of roots were determined. The results indicats that the disease of ginseng could be controlled better by the treatment B than by the treatment A within 45 days, but the better effect was achieved in the treatment A after storage for 45 days. Both storage methods had significant influence on the contents of ginsenoside, polysaccharide and free amino acids of roots. For the treatment A, the ginsenoside content of roots had remained relatively high during the storage period, and the crude polysaccharide and free amino acids changed slowly within 135 days and then increase significantly until 180 days. For the treatment B, the content of crude polysaccharide of roots decreased obviously after 90 days of storatge and then became stable until 135 days. The change of content of free amino acids was similar to the crude polysaccharide, but the decline was not significant. In general, the treatment A is more benefit to keep the quality of fresh roots of American Ginseng during 180 days of storage.
Amino Acids ; analysis ; Ginsenosides ; analysis ; Panax ; chemistry ; Plant Extracts ; analysis ; Plant Roots ; chemistry ; Technology, Pharmaceutical ; methods

Objective To observe the correlation between the changes of neural cell apoptosis arid caspase-3 gene expression in brain tissues following acute severe traumatic injury to brain(TIB).Method A total of 120 adult Spraque-Dawley rats were divided into a control group(n=8)，TIB group(n=56)and TIB with administration of caspase-3 inhibitor group(n=56).TIB models of rats were made with Feeney's method.The z-DEVDfmk(5 μg)，caspase-3 inhibitor，was administered by intracerebral infusion，and the rats were sacrificed 1，6，24，48 hours and 3，7，14 days postinjury(n=8 for each interval).The specimens of the injured cerebral cortex，suhcerticai white matter，hippocampus，dentate gyrus and contrahteral corresponding brain tissues were taken for detecting apoptesis of neural cells by the terminal deoxynucleotidyl transferase mediated DUTP nick end labeling (TUNEL)methods and flow cytomeay.Caspase-3 mRNA and protein expression were detected by using RT-PCR，immunohistochemistry and western blot analysis.The caspase-3 activity was detected by using caspase-3 fluorescent assay kit.Student t-test and Spearman correlation analysis were used to analyze the data with SPSS version 10.1 software package.Results Apoptesis indexes(AI)and the apoptesis percentage(AP)of neural cells in the injured brain regions increased quickly after injury，and reached its peak 24 to 48 hours later，then decreased slowly,but it remained at higher level above that of normal till 14 days later(P＜0.01).The levels of caspase-3 mRNA,eastme-3 protein and caspase-3 activity were increased significantly post injury，and reached its peak at 24 to 48 hours，then it gradually decreased.Compared with control group，the levels ofoptical density of caspase-3 proteins in the injured hippocampus and subcortical white matter at 24 and 48 hours post injury increased 1484% and 1690%，caspase-3 mRNA expressiom increased 1043%and 1180%，and the degreas of caspase-3 activity increased 148% and 183%，respectively.The expression of caspase-3 proenzyme and its P17 subarrit increased.After trealment with caspase-3 inhibitor z-DEVD-fmk，the levels of caspase-3 mRNA，protein expression and caspase-3 activity were significantly decreased.and AI and AP were significantly decreased as well.The correlation between caspase-3 mRNA and level of neural apoptesis was positive(r=0.821，P＜0.01)，and it was likewise between caspase-3 protein and level of neural apoptosis(r=0.638.P＜0.01).Interestingly enough，a positive correlation was found between caspase-3 mRNA and easpase-3 proteins(r=0.945，P＜0.01).Conclusions The activation of caspase-3 leads to apoptosis of neural cells after acute TIB.The expression of caspase-3 are consistent with apoptosis of neural cells following TIB.The regulation of caspase-3 induced by TIB occurs at a ceriain critical link before transduction.Caspase-3 inhibitor can efficiently inhibit apoptosis of neural cells following TIB.

Objective To investigate curative effects of hypothermia on the secondary axotomy of nondisruptive axonal injury (NDAI) after diffuse brain injury (DBI).Methods A total of 16 male Sprague-Dawley rats were randomly and equally divided into hypothermia group (at 32℃ for 6 hours) and control group (at 37.5℃ ).The axonal swelling and axonal balls were detected by means of NF68kD immunochemistry after DBI caused by fluid percussion.The changes of maximal density of axonal swelling and axonal balls in callosum,diencephalon-mesencephalon,pons-oblongata and cerebellum were compared 24 and 72 hours after injury between both groups.Results NF68kD immunochemistry well showed axonal swellings and axonal balls in whole brain.The axonal swelling and axonal balls were significantly decreased 24 hours after DBI in both groups (P<0.05),especially in diencephalon-mesencephalon ,pons-oblongata and cerebellum (P<0.01).While there showed significant decrease of axonal swellings and axonal balls in pons-oblongata and cerebellum in hypothermia group 72 hours after DBI (P<0.05,P<0.01) but insignificant changes in the callosum and the diencephalon-mesencephalon compared with control group (P>0.05 ).Conclusions Hypothermia can retard the progress of mild or severe NDAI at early stage,which would taper with the longer time after injury except for partial mild NDAI.Hypothermia may prevent mild NDAI from secondary axotomy.

Objective To locally inject human umbilical cord blood (HUCB) mesenchymal stem cells (MSCs) to rat traumatic brain injury (TBI) model to investigate expression of neural markers and neurological functional improvement. Methods HUCB-MSCs were labeled by bis-benzimide for over 24 hours and stereotactically transplanted into the brain of the rats. All rats were divided into four groups, ie, sham injury group, TBI group, control (TBI + PBS) group and treatment (TBI + MSCs) group, Im-munohistochemical methods and immanofluorescence staining were used to observe the survival, migration and differentiation of the transplanted cells. The neurological functional improvement was evaluated by u-sing the neurological severity score (NSS). Results There existed a large number of MSCs survived in local region of the brain that received transplants, when some MSCs differentiated into neurons or astro-cytes and expressed the neurocyte markers including NSE and GFAP around the grafted site. Treatment group had significantly improved scores compared with sham injury group, TBI group and control group. Conclusions HUCB-MSCs transplantation can potentially improve neurological functional after TBI and may be a good alternative to bone marrow cells for stem cell transplantation or cell therapy.