Abstract: Objective　In order to explore the application prospects of the phenyl pyrazole insecticide fipronil for mosquito control and identify potential target genes involved in the resistance of Aedes aegypti to fipronil, and lay the foundation for an in-depth study of the resistance mechanism of Aedes aegypti to fipronil. Methods　Using Aedes aegypti sensitive strains as experimental materials, Aedes aegypti larvae were treated with fipronil, and the differences in gene expression of Aedes aegypti larvae before and after drug administration were compared at the transcriptome level using transcriptome sequencing combined with bioinformatics analysis, and the differential genes were analyzed. Results　A total of 757 differentially expressed genes were identified between the fipronil-treated group and control group, including 217 and 540 up- and down-regulated genes, respectively. Among these, the expression of glutamate-gated chloride channel (GluCls) genes varied significantly before and after treatment. Gene ontology analysis revealed that differentially expressed genes were enriched in catalytic activity, binding, metabolic processes, and membrane-related functions, while KEGG pathway analysis indicated enrichment in biosynthesis, metabolism, and life regulation processes, while the glutathione metabolic pathway was enriched in 15 differentially expressed genes. Conclusions The transcriptome results revealed that GST gene expression was significantly upregulated in fipronil-treated Aedes aegypti larvae, indicating that GST gene is involved in the development of fipronil resistance in Aedes aegypti larvae. In addition, GluCls gene expression was also significantly different before and after treatment, suggesting that GluCls migh be a potential target receptor for fipronil resistance in Aedes aegypti. As GluCls is an ideal target receptor found only in invertebrates, this discovery provides a reference and basis for further exploration of the toxicological mechanism of fipronil on Aedes aegypti.

AIM To eslablish an HPLC method for the simultaneous content determination of brazilin,(±) protosappanin B,hydroxysafflor yellow A,safflor yellow A,oxypeucedanin,imperatorin and isoimperatorin in Waicha Bailing Tincture (Angelicae dahuricae Radix,Sappan Lignum,Carthami Flos,etc.).METHODS The analysis of methanol extract of this drug was performed on a Agilent TC-C18 column (200 mm ×4.6 mm,5 μm),with the mobile phase comprising of methanol-0.1％ glacial acetic acid flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelengths were set at 285,403,310 nm.RESULTS Seven constituents showed good linear relationships within their own ranges (r ≥0.999 5),whose average recoveries were 96.91％-99.59％ with the RSDs of 0.81％-1.19％.CONCLUSION This stable and reliable method can be used for the quality control of Waicha Bailing Tincture.

OBJECTIVETo analyze the relationship between hepatitis B virus (HBV) precore (PC) and basal core promoter (BCP) mutations and HBV-related acute-on-chronic liver failure (HB-ACLF).

METHODSForty-four patients with HB-ACLF and 28 patients with chronic hepatitis B (CHB; used as controls) were enrolled and venous blood samples were collected from all individuals. The PC and BCP gene fragments were amplified by nested PCR. HBV genotype and BCP/PC mutations were determined by direct sequencing and analysis by BioEdit (version 7.0.9.0). Ten of the HB-ACLF patients were selected for follow-up (range: 2-8 weeks), which included once weekly sera collection to determine the relation of mutations and treatment response. Serum levels of HBV DNA were measured by real-time PCR assay, and alanine aminotransferase, total bilirubin, creatinine and albumin were measured by standard biochemical assay and used to determine the MELD score.

RESULTSAll 44 HB-ACLF patients were infected with HBV genotype C. In the CHB group, 26 patients were infected with genotype C and two with genotype B. Single mutations (A1762T, G1764A, T1753V, G1896A, and G1899A) and combined mutations (A1762T + G1764A, G1896A + G1899A, T1753V+ A1762T + G1764A, G1896A + G1899A + A1762T + G1764A, and A1762T + G1764A + G1896A) were more frequently detected in HB-ACLF patients than in CHB patients (P less than 0.05). A significantly higher proportion of PC/BCP wild-type sequences was found in patients with CHB than in patients with HB-ACLF (17.9% vs. 2.3%; x² = 5.440, P = 0.020). The proportion of patients carrying both PC and BCP mutations was significantly higher in HB-ACLF patients than in CHB patients (79.6% vs. 39.3%; x² = 12.021, P = 0.001). The proportion of patients carrying only BCP mutation was 42.9% in the CHB group and 20.5% in the HB-ACLF group (x² = 4.157, P = 0.041). No occurrences of only PC mutation were detected in either the CHB or HB-ACLF group. The combined mutations were present in all 10 of the HB-ACLF follow-up patients. Mutations G1899A, T1753V, and A1846T were correlated with disease recovery. Significant decreases in the MELD score were accompanied by decreases in the A1846T mutation.

CONCLUSIONSignificantly more HB-ACLF patients carried HBV with mutations in the PC and BCP than CHB patients. Moreover, more HB-ACLF patients carried HBV with PC + BCP combined mutations and PC mutation only. The G1899A, T1753C, and A1846T mutations were associated with HB-ACLF response to treatment and improvement in liver function.

Adult ; Case-Control Studies ; DNA, Viral ; genetics ; End Stage Liver Disease ; Female ; Genetic Variation ; Genotype ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Liver Failure ; virology ; Male ; Middle Aged ; Mutation ; Promoter Regions, Genetic

The reaction conditions of baicalin hydrolyzed into baicalein by a kind of thermophilic and sugar-tolerant beta-glucosidase were studied in this paper. The beta-glucosidase could catalyze baicalin into baicalein well in the acetic acid-sodium acetate buffer. The optimal enzyme activity was at 85 degrees C and pH 5.5. The enzyme was stable at the temperature less than 85 degrees C and pH range of 5-7.5. The maximum reaction rate V. and michaelis constant K. were 0.41 mmol x L(-1) x min(-1) and 3.31 mmol x L(-1) respectively. Different metal ions had different effects on the activity of enzyme. Na+ existing in acetic acid-sodium acetate buffer had an activation effect on enzyme. The enzyme activity was enhanced by the concentrations of glucose below 0.6 mol x L(-1), and was gradually inhibited when monosaccharide concentration was over 0.6 mol x L(-1). When the monosaccharide concentration reached 1.2 mol x L(-1), the inhibition rate of enzyme activity was about 50%, which showed good glucose tolerance. The good reaction conditions through the experiment have been determined as follows, the substrate: enzyme dose was 1 g: 0.2 mL, acetic acid-sodium acetate buffer pH 5.5, reaction temperature 85 degrees C, reaction time 10 h, and the enzymatic hydrolyzation ratio could reach 97%.
Biocatalysis ; Enzyme Stability ; Flavanones ; chemistry ; Flavonoids ; chemistry ; Glucose ; chemistry ; Hot Temperature ; Hydrolysis ; Kinetics ; beta-Glucosidase ; chemistry

OBJECTIVETo explore the molecular mechanism of APL cell resistance to ATRA.

METHODSThe ATRA sensitive and resistant APL cell lines, NB4 and NB4-R1, were used as in vitro models. The effects of specific inhibitors and activators of adenylate cyclase (AC) and phosphodiesterase (PDE) on ATRA-induced differentiation was evaluated by cell morphology, cell surface antigen expression and nitroblue-tetrazolium (NBT) reduction assays.

RESULTSSQ22536, a specific antagonist of AC, could dramatically block ATRA-induced NB4 cell differentiation. When ATRA + SQ22536 group compared with ATRA group, the positivity of CD11b decreased from (95.9 +/- 2.5)% to (60.3 +/- 7.1)%, while the A(540) in NBT reduction assay decreased from 0.585 +/- 0.092 to 0.170 +/- 0.028 (P < 0.05). Forskolin, an agonist of AC, could overcome the resistance of NB4-R1 cells to ATRA. When ATRA + forskolin group compared with ATRA group, the positivity of CD11b increased from (34.3 +/- 5.3)% to (94.6 +/- 2.4)%, while the A(540) in NBT reduction assay increased from 0.110 +/- 0.028 to 0.395 +/- 0.049 (P < 0.05). In contrast, the specific antagonist and agonist of PDE, 3-isobutyl-1-methylxanthine (IBMX) and calmodulin, exerted little impact on ATRA treatment.

CONCLUSIONSThe defaults in the initiation of AC activation may contribute to the resistance to ATRA in some APL cells.

Adenine ; analogs & derivatives ; pharmacology ; Adenylyl Cyclase Inhibitors ; Adenylyl Cyclases ; metabolism ; Antineoplastic Agents ; pharmacology ; CD11b Antigen ; metabolism ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Activation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Phosphoric Diester Hydrolases ; metabolism ; Tretinoin ; pharmacology

To set up a new pattern of quality control and evaluation for Chinese medicine. By investigating the limitation of quality control pattern for Chinese medicine, the differences and similarities in the chemical substantial style as well as quality control patterns among Chinese medicine, chemical synthetic drugs and Biologicals, combining with the author's experience on the research of geo-authentic medicinal material and theory of Chinese medicinal nature, a new pattern of quality control for Chinese medicine has been explored and designed. A more rational pattern of quality control for Chinese medicine should be referred to Biologicals instead of chemical synthetic drugs, there are more similarity in chemical substantial style and quality control pattern for Chinese medicine between Chinese medicine and Biologicals than that between Chinese medicine and chemical synthetic drugs. Based on geo-authentic medicinal material and bioassay or biopotency detection, a new pattern of quality control for Chinese medicine could be built and applied.
Biological Assay ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Energy Transfer ; Evaluation Studies as Topic ; Medicine, Chinese Traditional ; standards ; Pattern Recognition, Automated ; Plants, Medicinal ; chemistry ; Quality Control ; Technology, Pharmaceutical ; methods

Chinese medicinal authentication is fundamental for the standardization and globalization of Chinese medicine. The discipline of authentication addresses difficult issues that have remained unresolved for thousands of years, and is essential for preserving safety. Chinese medicinal authentication has both scientific and traditional cultural connotations; the use of scientific methods to elucidate traditional experience-based differentiation carries the legacy of Chinese medicine forward, and offers immediate practical significance and long-term scientific value. In this paper, a path of inheritance and innovation is explored through the scientific exposition of Chinese medicinal authentication, featuring a review of specialized publications, the establishment of a Chinese medicine specimen center and Chinese medicinal image databases, the expansion of authentication technologies, and the formation of a cultural project dedicated to the Compedium of Materia Medica.
Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; standards ; Humans ; Materia Medica ; chemistry ; standards ; Medicine, Chinese Traditional ; standards ; Reference Standards

BACKGROUNDThe auditory brainstem implants (ABIs) have been used to treat deafness for patients with neurofibromatosis Type 2 and nontumor patients. The lack of an appropriate animal model has limited the study of improving hearing rehabilitation by the device. This study aimed to establish an animal model of ABI in adult rhesus macaque monkey (Macaca mulatta).

METHODSSix adult rhesus macaque monkeys (M. mulatta) were included. Under general anesthesia, a multichannel ABI was implanted into the lateral recess of the fourth ventricle through the modified suboccipital-retrosigmoid (RS) approach. The electrical auditory brainstem response (EABR) waves were tested to ensure the optimal implant site. After the operation, the EABR and computed tomography (CT) were used to test and verify the effectiveness via electrophysiology and anatomy, respectively. The subjects underwent behavioral observation for 6 months, and the postoperative EABR was tested every two weeks from the 1 st month after implant surgery.

RESULTThe implant surgery lasted an average of 5.2 h, and no monkey died or sacrificed. The averaged latencies of peaks I, II and IV were 1.27, 2.34 and 3.98 ms, respectively in the ABR. One-peak EABR wave was elicited in the operation, and one- or two-peak waves were elicited during the postoperative period. The EABR wave latencies appeared to be constant under different stimulus intensities; however, the amplitudes increased as the stimulus increased within a certain scope.

CONCLUSIONSIt is feasible and safe to implant ABIs in rhesus macaque monkeys (M. mulatta) through a modified suboccipital RS approach, and EABR and CT are valid tools for animal model establishment. In addition, this model should be an appropriate animal model for the electrophysiological and behavioral study of rhesus macaque monkey with ABI.

Animals ; Auditory Brain Stem Implants ; Deafness ; surgery ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Macaca mulatta ; Male

OBJECTIVETo synthesize cyclin-dependent kinase (CDKs) inhibitors and assay their antitumor activities.

METHODSA series of pyrimidines containing different arylamino and 1-(methylsulfonyl)piperidin moieties were designed by combining the segments 1-(methylsulfonyl)piperidin and pyrimidine heterocycles according to the super-position principle of the reinforcement of biological activities.

RESULTSTheir structures were characterized by MS and 1H NMR spectra and all the synthesized compounds were screened for their antimicrobial activity with MTT assay.

CONCLUSIONThe preliminary bioassay showed that compound 3 b displayed good antitumor activity (IC(50)=13.6 µmol/L). The preliminary structure activity relationship analysis of these analogues suggest that the steric factor may have important impact on the anti-tumor activity.

Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Humans ; Pyrimidines ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship

Recently, some progress has been made in the clinical study of obsessive compulsive disorder (OCD). Human research has inherent limitation because of its non-invasive nature, while animal models can break through the shortcomings of human research, allowing us to more accurately study the pathophysiology of OCD. Therefore, many researchers tried to use genetic knockout animal models to further study the etiology of OCD. In this review, recent progress about this kind of research was reviewed.