AIM To study endophytic fungi from Scutellaria baicalensis Georgi.and the enzyme inhibitory activities of their secondary metabolites.METHODS Six different media were used to isolate and purify endophytic fungi from S.baicalensis by tissue homogenate method.The activities of secondary metabolites were evaluated by targeting different enzymes.The highly active strains were identified by molecular biology combined with morphology,and the highly active chemical components were tracked and separated by modern chromatographic separation technology.RESULTS Sixty-four endophytic fungal strains were isolated from S.baicalensis,and one hundred and twenty-eight secondary metabolites were obtained by fermentation.The samples with certain inhibitory activities against adenosine deaminase(ADA),β-lactamase and tyrosinase(TYR)accounted for 14.06%,3.91%and 18.75%,respectively.Strain HTS-23-2 showed high TYR inhibitory activity,and 99%homology with Aspergillus flavus by molecular identification.One compound was isolated from the fermentation samples and identified as kojic acid.CONCLUSION S.baicalensis harbors a rich diversity of endophytic fungi,which serve as a valuable resource for active substances.

Enzymes are closely associated with the onset and progression of numerous diseases, making enzymes a primary target in innovative drug development. However, the challenge remains in identifying compounds that exhibit potent inhibitory effects on the target enzymes. With the continuous expansion of the total number of natural products and increasing difficulty in isolating and enriching new compounds, traditional high-throughput screening methods are finding it increasingly challenging to meet the demands of new drug development. Virtual screening, characterized by its high efficiency and low cost, has gradually become an indispensable technology in drug development. It represents a prominent example of the integration of artificial intelligence with biopharmaceuticals and is an inevitable trend in the rapid development of innovative drug screening in the future. Therefore, this article primarily focused on systematically reviewing the recent applications of virtual screening technology in the development of enzyme inhibitors and explored the prospects and advantages of using this technology in developing new drugs, aiming to provide essential theoretical insights and references for the application of related technologies in the field of new drug development.
Artificial Intelligence ; Enzyme Inhibitors/pharmacology* ; High-Throughput Screening Assays ; Molecular Docking Simulation

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases

Objective To investigate the curative effect of YU Yun pulse-feeling-based acupuncture therapy for the treatment of middle-late liver cancer. Methods A total of 60 middle-late liver cancer patients were divided into control group and treatment group by stratified randomization method,30 cases in each group. The control group was given integrated Chinese and western medicine therapy according to the clinical pathway, including anti-cancer treatment such as vascular interrention,molecular targeted therapy,chemotherapy,radio therapy, and focal ablation therapy,as well as chinese medicine treatment based on disease differentiation and syndrome differentiation. And the treatment group was given YU Yun pulse-feeling-based acupuncture therapy on the basis of treatment for the control group. The two groups received 12-week treatment, and then their curative effects were compared. Results The treatment group had better effect on increasing the survival rate, prolonging survival time, improving the scores of clinical symptoms, stabilizing tumor size, and increasing the scores of Karnofsky Performance Status (KPS)than the control group,the difference being significant (P < 0.05 or P <0.01). Conclusion YU Yun pulse-feeling-based acupuncture therapy exerts certain curative effect for the treatment of middle-late liver cancer.

OBJECTIVETo screen and verify differentially expressed genes in prostate cancer.

METHODSUsing DNA microarray, we screened differentially expressed genes in prostate cancer tissue and its adjacent tissue followed by verification by PCR.

RESULTSA total of 1 444 genes were found to be differentially expressed (differentiation ≥ 1.5-fold; P≤ 0.05) in the prostate cancer tissue, of which 769 (53%) were up-regulated and 675 (47%) down-regulated. Fifty percent of the differentially expressed genes showed a 1.5- to 2-fold differentiation, including 396 up-regulated and 182 down-regulated ones. Additionally, 308 up-regulated and 334 down-regulated genes exhibited a >2- to 5-fold, 46 up-regulated and 78 down-regulated genes a > 5- to 10-fold, and 19 up-regulated and 81 down-regulated genes a > 10-fold differentiation. Verification by subjecting 15 most significantly up-regulated and another 15 most markedly down-regulated genes to quantitative real-time PCR (qRT-PCR) showed that most of the genes had a transcriptional profile similar to that in the microarray data, with a Pearson correction coefficient of 0.83 between the microarray data and qRT-PCR results. Totally, 10 significantly differentially expressed genes were identified.

CONCLUSIONDNA microarray analysis provides reliable information on differentially expressed genes in prostate cancer and benign tissues. The 10 significantly differentially expressed genes verified by qRT-PCR could possibly become new bio-markers and specific molecules for tumor identification.

Cell Differentiation ; Down-Regulation ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Prostatic Neoplasms ; genetics ; Transcriptional Activation ; Up-Regulation

Postoperative radiotherapy is a major treatment for patients with maxillary sinus carcinoma. However, the irregular resection cavity poses a technical difficulty for this treatment, causing uneven dose distribution to target volumes. In this study, we evaluated the dose distribution to target volumes and normal tissues in postoperative intensity-modulated radiotherapy (IMRT) after placing a water-filled balloon into the resection cavity. Three postoperative patients with advanced maxillary sinus carcinoma were selected in this trial. Water-filled balloons and supporting dental stents were fabricated according to the size of the maxillary resection cavity. Simulation CT scans were performed with or without water-filled balloons, IMRT treatment plans were established, and dose distribution to target volumes and organs at risk were evaluated. Compared to those in the treatment plan without balloons, the dose (D98) delivered to 98% of the gross tumor volume (GTV) increased by 2.1 Gy (P = 0.009), homogeneity index (HI) improved by 2.3% (P = 0.001), and target volume conformity index (TCI) of 68 Gy increased by 18.5% (P = 0.011) in the plan with balloons. Dosimetry endpoints of normal tissues around target regions in both plans were not significantly different (P > 0.05) except for the optic chiasm. In the plan without balloons, 68 Gy high-dose regions did not entirely cover target volumes in the ethmoid sinus, posteromedial wall of the maxillary sinus, or surgical margin of the hard palate. In contrast, 68 Gy high-dose regions entirely covered the GTV in the plan with balloons. These results suggest that placing a water-filled balloon in the resection cavity for postoperative IMRT of maxillary sinus carcinoma can reduce low-dose regions and markedly and simultaneously increase dose homogeneity and conformity of target volumes.
Adult ; Aged ; Carcinoma, Adenoid Cystic ; diagnostic imaging ; radiotherapy ; surgery ; Carcinoma, Squamous Cell ; diagnostic imaging ; radiotherapy ; surgery ; Female ; Humans ; Male ; Maxillary Sinus ; surgery ; Maxillary Sinus Neoplasms ; diagnostic imaging ; radiotherapy ; surgery ; Middle Aged ; Postoperative Period ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated ; methods ; Stents ; Tomography, X-Ray Computed

Based on the duck plague virus (DPV) UL35 gene sequence that our laboratory obtained (GenBank accession number EF643558), a pair of primers was designed using Oligo6.0 and primer5.0, then the UL35 gene was amplified from DPV CHv strain genomic DNA and cloned into the pMD18-T to construct a clone plasmid pMD18-T-UL35. After identification of the pMD18-T-UL35 by PCR amplification and restriction digestion, the fragment of the UL35 gene was subcloned into the prokaryotic expression vector pET-32a(+). The resultant recombinant plasmid pET-32a(+)-UL35 was then transformed into E. coli BL21 (DE3) strain and optimally-expressed under the induction of 1.0 mmol/L IPTG at 34 degrees C for 5 hours. SDS-PAGE analysis showed the recombinant protein (VP26) had a molecular weight of about 33KDa and accounted for 32.3% of total bacterial protein by gel scanning. The protein was then purified by Ni(2+)-affinity chromatography and used to immunize rabbit for producing the VP26 anti-serum and its antibody titer was up to 1:32 detected by agar diffusion reaction. After the IgG of the polyclonal antibodies was purified by High-Q anion-exchange chromatography, Western blot analysis indicated that the IgG had specific reaction with the VP26. Moreover, the subcellular localization detection was observed using immunofluorescence technique. The results showed that the specific fluorescences appeared relatively few in nucleus in 2 to 8 hours and increased gradually in 12 to 36 hours and eventually reached to the maximum, which aggregated in the spot region of the nucleus after the duck embryo fibroblast (DEF) were infected by DPV. However, there were only a small amount of specific fluorescences in the cytoplasm in 12 hours and increased with the extension of infection time in 24 to 48 hours. The specific fluorescences finally reached to the maximum in the cytoplasm in 72 hours. The results provided significant data for furthering the study on the function of DPV UL35 gene.
Animals ; Blotting, Western ; Capsid Proteins ; chemistry ; genetics ; metabolism ; Cell Nucleus ; metabolism ; Cells, Cultured ; Cloning, Molecular ; Ducks ; virology ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Fibroblasts ; cytology ; metabolism ; virology ; Herpesviridae ; genetics ; metabolism ; Microscopy, Fluorescence ; Molecular Weight ; Plasmids ; genetics ; Polymerase Chain Reaction ; Rabbits ; Recombinant Proteins ; genetics ; immunology ; metabolism

Objective To explore the existence of natural loci on Marmota himalayana plague in Sichuan province and to provide basis for prevention and control of the disease. Methods Both epidemiological investigation and laboratory tests were used to provide the host animal and fleas of the vectors with Yersinia pestis carriers. Results 30 species of animals were found to belong to 10 orders. Ochotona curzoniae and M.himalayana were the most common ones while 7 species of the fleas belonged to 7 genera and 3 families. M.himalayana was the main reservoirs while Callopsylla dolabris and Oropsylla silantiewi served as vectors. The 13 Y.pestis were identified from 43 Marmota samples. 8 samples were identified under IHA, with the highest titer of herding-dogs serum as 1 : 10 240. 19 samples were F1 antigen positive using RIHA and the highest titer of M.himalayana serum was 1:409 600. The major foci was 4545 km2, distributed at Dege county in Sichuan province. Conclusion We have confirmed the existence of natural foci on M. Himalayana plague in Sichuan province.

Background and Objective:The gross tumor volume (GTV) obviously reduces after induction chemotherapy (IC) for primary Iocoregionally advanced nasopharyngeal carcinoma (NPC). This study was to investigate the impact of changing gross tumor volume delineation on the dose distribution and clinical treatment outcome after IC. Methods: From January 2008 to April 2009, 24 patients with Stage Ⅲ-Ⅳb primary locoregionally advanced NPC were treated with TPF regimen IC followed by intensitymodulated radiotherapy (IMRT) with concurrent chemotherapy The primary GTVs were delineated into two parts: the post-IC primary GTV (GTVpost-IC-NP),and the region of pre-IC primary GTV minus GTVpost-IC-NP (GTVprepost-IC-NP). The dose distributions of two plans with GTVpost-IC-NP or pre-IC primary GTV were assessed by analyzing ten cases. The clinical treatment outcome and toxicity of all patients were observed. Results:The post-IC GTV was significantly smaller than the pre-IC GTV(primary GTV 25.5 cm~3 vs.51.1 cm~3,P=0.001;lymph nodes GTV 9.1 cm~3 vs. 31.4 cm~3, P=0.035;primary+lymph nodes GTV 33.2 cm~3 vs. 82.6 cm~3, P=0.004), the overall GTV with an average shrinkage of 61%. The high dose region was also smaller after IC(volumes covered by 64.4 Gy were 422.9 cm~3 vs.457.9cm~3, P=0.003; 274.2 cm~3 vs.334.5 cm~3 by 68 Gy, P=0.041). The complete response rate was 38% after IC,and 100% three month after radiotherapy.The toxicity of following IMRT with concurrent chemotherapy was similar to that of IMRT with concurrent chemotherapy alone. With median follow-up of 9 months, the locoregionally control rate was 100% and only one patient presented metastasis 15 months after treatment. Gonclusions: TPF regimen IC could significantly reduce tumor volume. The following IMRT with GTVpost-IC-NP plan reduced the high dose region,which didn't add toxicity while had excellent short-term treatment outcome.

OBJECTIVETo screen the polypeptides specifically binding to human large intestinal cancer LoVo cells from a phage-displayed peptide library for potential use as targeting vectors for large intestinal cancer therapy.

METHODSWith the LoVo cells as the target cells and human normal large intestinal mucosal epithelial cells as the absorber cells for subtraction biopanning from a c7c phage-display peptide library, the positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence detection. The amino acid sequences of the identified peptides were deduced by DNA sequencing.

RESULTSAfter 3 rounds of screening, 5 positive phage clones showing specific binding to LoVo cells and containing conserved motif RPMP were obtained from the 20 randomly selected clones.

CONCLUSIONSpecific peptide against large intestinal cancer cells can be obtained from a phage-display peptide library for use as potential vectors for targeting therapy of large intestinal cancer.

Amino Acid Sequence ; Base Sequence ; Binding, Competitive ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Molecular Sequence Data ; Peptide Library ; Peptides ; genetics ; isolation & purification ; metabolism ; Protein Binding