Adult ; Aged ; Aged, 80 and over ; Bile Duct Diseases ; diagnosis ; Bile Duct Neoplasms ; diagnosis ; Cholangiopancreatography, Endoscopic Retrograde ; Cytodiagnosis ; Cytological Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Diseases ; diagnosis ; Pancreatic Neoplasms ; diagnosis

OBJECTIVETo observe mainfestations of syndrome and biochemical indices of hypertensive model rats with excessive accumulation of phlegm-dampness syndrome (EAPDS), and to explore its possible pathological mechanism.

METHODSEAPDS rat model was prepared in 50 Wistar rats by feeding with high fat forage. Meanwhile, a normal control group consisting of 10 Wistar rats was set up by feeding with normal forage. After 25-week continuous feeding, 22 rats with body weight (BW) and blood pressure (BP) exceeding 25% those of the control group were selected as a model group. BW, BP, blood lipids, and related serological indicators were detected in all rats. Morphological changes of target organs were observed. mRNA expression levels of leptin receptor (LepR), Janus kinase2 (Jak2), signal transducer and activator of transcription 3 (Stat3), suppressor of cytokine signaling-3 (Socs3), angiotensin II receptor type 1 (AT1), angiotensin II receptor type 2 (AT2), phosphatidylinositol 3 kinase (P13K), serine threonine kinase (Akt), nuclear factor of kappa B (NF-κBp65), inhibitor of nuclear factor kappa-B kinase α (IKKα), NF-kappa-B inhibitor β (lKKβ), NF-kappa-B inhibitor α (IKBα), and AMP-activated protein kinase (AMPK) were detected by quantitative real-time PCR (qPCR). Expression levels of AT1 and LepR in aorta were detected by immunohistochemical assay and Western blot respectively.

RESULTSCompared with the control group, BW, BP, and blood lipids increased; serum levels of leptin (Lep) , Ang II, Hcy, ET-1, TNF-α, IL-6, and p2-MG increased, but NO decreased in the model group (P < 0.05, P < 0.01). Aortal endothelial injury and smooth muscle cell proliferation occurred in the model group, accompanied with heart and renal injury. Compared with the control group, mRNA expression levels of LepR, Jak2, Stat3, Socs3, AT1 , PI3K, Akt, NF-κB p65, IKKβ, IKBα, and AMPK in aorta were up-regulated significantly (P < 0.05), while the expression of IKKa decreased (P < 0.05). Immunohistochem- ical staining showed, brownish yellow deposit of AT1 and LepR was obviously increased, with more extensively positive distribution. Western blot results showed, as compared with the control group, protein expression levels of AT1 and LepR obviously increased in the model group (P < 0.05).

CONCLUSIONSModel rats exhibited typical syndromes of EAPDS. They put up weight with fat abdomen, gloomy hair, poor appetite, hypersomnia, lowered activities , reduced food intake, loose stool, dark red tongue, white tongue with white, thick, greasy fur. Lep could be taken as one of objective indicators for evaluating hypertension rat model with EAPDS.

Animals ; Aorta ; Cell Proliferation ; Disease Models, Animal ; Hypertension ; physiopathology ; I-kappa B Proteins ; Interleukin-6 ; Leptin ; blood ; NF-KappaB Inhibitor alpha ; NF-kappa B ; Phosphatidylinositol 3-Kinases ; Rats ; Rats, Wistar ; Suppressor of Cytokine Signaling Proteins ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha

Central composite design (CCD) is one of the most commonly used design methods in response surface optimization and has been widely applied in the field of pharmaceutics to optimize preparations. On the 20th anniversary of the introduction of CCD into China, the paper reviews its application in domestic pharmaceutical researches. Based on the brief introduction of basic principle and operation steps of CCD, the mistakes emerging in the application of CCD are summarized, including conceptual confusion with Box-Behnken design and face-centered CCD as well as wrong designs. Besides, the issues concerning the selection of factors and responses are discussed. The article is helpful for researchers to comprehensively understand the CCD and facilitates the rational application of this method.

In recent years, artificial intelligence (AI) technology has advanced rapidly and has been widely applied in various fields such as medicine and pharmacy, accelerating the drug development process. Focusing on the application of AI in the discovery and optimization of lead compounds, this review provides a detailed introduction to AI-assisted virtual screening and molecular generation methods for discovering lead compounds, while particularly highlighting the cases of AI-drived drugs into clinical trials. Additionally, we briefly outline the application of AI basic algorithm models in quantitative structure-activity relationship (QSAR) and drug repurposing, offering insights for AI-based drug discovery.

The aim of this study was to investigate the expression of matrix metalloproteinase 26 (MMP-26), tissue inhibitor of metalloproteinase-4 (TIMP-4) and matrix metalloproteinase 9 (MMP-9) in patients with diffuse large B cell lymphoma (DLBCL) and their correlations with pathogenesis and development of DLBCL. A total of 95 specimens excised from DLBCL patients were prepared. Expression of MMP-26, TIMP-4 and MMP-9 were tested by SABC immunohistochemistry method and its correlation to clinicopathology indexes were analyzed. The results showed that as compared with reactive hyperplasia of lymph nodes, the high expression of MMP-26, TIMP-4 and MMP-9 were found in different types of DLBCL. The positive expression rate of MMP-26 was related to immune typing (P < 0.05). The expression level of MMP-26 in GCB was lower than that in non-GCB, and did not relate to clinical staging, age, sex, diseased region (P > 0.05). The positive expression rate of MMP-9 was related to clinical staging, the positive expression rate of MMP-9 proteins in patient at III and IV stage was obviously higher than that in patients at I and II stage, but did not relate to immune type, age, sex and diseased region of DLBCL (P > 0.05). The expression of TIMP-4 did not relate to immune type, clinical stage, age, sex, disease region (P > 0.05). The expression of MMP-26 in pathologic tissue of DLBCL did not relate to expression of TIMP-4, but positively related to expression of MMP-9 protein (r = 0.486, P < 0.05). It is concluded that MMP-26 and MMP-9 synergically express in DLBCL. MMP-26 may be involve in pathogenesis and invasiveness of DLBCL, the expression of MMP-26 relates to subtypes of DLBCL. The MMP-26 may serve as an indicator for typing of DLBCL and contributes to predict the invasion and metastasis of DLBCL and itself may become a potential target for therapy.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinases, Secreted ; metabolism ; Middle Aged ; Tissue Inhibitor of Metalloproteinases ; metabolism ; Young Adult

OBJECTIVETo construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.

METHODSFull length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope.

RESULTSThe recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.

CONCLUSIONdelta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.

Animals ; DNA, Complementary ; genetics ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVEOptimization of combining electroencephalography (EEG), short latency somatosensory evoked potentials (SLSEP) and transcranial Doppler (TCD) techniques to diagnose brain death.

METHODSOne hundred and eleven patients (69 males, 42 females) from the major hospitals of Zhejiang Province were examined with portable EEG, SLSEP and TCD devices. Re-examinations occurred < or =12 h later.

RESULTSThe first examination revealed that the combination of SLSEP and EEG led to more sensitive diagnoses than the combination of SLSEP and TCD. Re-examination confirmed this and also revealed that the combination of TCD and EEG was the most sensitive.

CONCLUSIONThe results show that using multiple techniques to diagnose brain death is superior to using single method, and that the combination of SLSEP and EEG is better than other combinations.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Brain Death ; diagnosis ; diagnostic imaging ; Child ; Electroencephalography ; methods ; Evoked Potentials, Somatosensory ; Female ; Humans ; Male ; Middle Aged ; Ultrasonography, Doppler, Transcranial ; methods ; Young Adult

Objective To evaluate the feasibility of making the osteoarthritis (OA) model in the medial collateral ligament and the medial meniscus excision in rats,and to explore the mechanism of MMP-13 and ADAMTS-5 protein in the cartilage of rat osteoarthritis models.Methods Forty SD rats were randomly divided into model group(n =30) and sham operation group(n =10).The knee joint OA model was made from the medial collateral ligament of the knee and the medial meniscus,and the sham operation group was sutured after opening the capsule of the knee joint.Rats in the model group were killed at 4,6,and 8 weeks after the operation,and the sham operation group died of the rats at 8 weeks after the operation.The articular cartilage tissue of rats was taken.The expression level of MMP-13 and ADAMTS-5 protein in cartilage tissue was detected by Western blot and immunohistochemistry.Results Cartilage degeneration was observed in the model group 4 weeks after operation,and degeneration was further aggravated at 6 and 8 weeks.There was no significant degeneration in the sham operation group.There was a significant difference in the articular cartilage score between the two groups (P ＜ 0.05).The results of Western blot detection showed that the protein expression of MMP-13 and ADAMTS-5 was low in the sham operation group.The MMP-13 model began to rise for 4 weeks,and continued to rise in the 6th week,and the 8th week was lower than that of the previous one.The protein expression had significant difference in all groups at 4 weeks,6 weeks and 8 weeks (P ＜ 0.05).The ADAMTS-5 model began to rise for 6 weeks,and the expression level was maintained before the model was maintained for 8 weeks.The protein expression at 4 weeks compared to that at 6 weeks and 8 weeks had significant difference(P ＜0.05).The protein expression in rat articular cartilage tissue at 6 weeks and 8 weeks had no significant difference (P ＞ 0.05).The expression trend of immunohistochemical detection protein was consistent with that of Western blot.Conclusion The animal model of OA can be established by using the method of medial collateral ligament dissection and medial meniscectomy.The expression level of MMP-13 and ADAMTS-5 in different stages of OA has changed significantly.Further research is needed to explore its related signaling pathways.

OBJECTIVE: To study the relationship between postmortem interval and the myofibril fragmentation index of skeletal muscle. METHODS: Rabbit skeletal muscle were left in the room temperature for different postmortem intervals, and the protein concentration of each sample was detected by using biuret method. Furthermore, the myofibril fragmentation index of each sample was measured under the protein concentration level of 0.5 mg/mL. RESULTS: The myofibril fragmentation index increased obviously according to the postmortem interval prolongation. CONCLUSION The myofibril fragmentation index may be used on the estimation of early postmortem interval.
Animals ; Calcium-Binding Proteins/metabolism* ; Female ; Male ; Muscle, Skeletal/metabolism* ; Myofibrils/metabolism* ; Postmortem Changes ; Proteins/metabolism* ; Rabbits ; Spectrophotometry ; Time Factors

Objective To establish the national quantity standard of hepatitis B surface antigen according to the world health organization's standard material and prepare, the national liner reference panel for hepatitis B surface antigen. Methods Sera from hepatitis B patients and health blood donors in different areas were collected and detected by domastic HBsAg kits, anti-HBs kits, HBeAg kits, anti-Hbe kits, anti-Hbe kits and anti-HCV,and then confirmed by the kits produced by Abbott, which was approved by WHO. One serum with high concentration of HBsAg was calibrated with the standard sample of WHO. And then it was diluted by 1.5 fold as the liner HBsAg reference panel. Results The HBsAg concentration of one serum was 1226 IU/ml calibrated by 21 independent standardization measurements with 7 kinds of kits. The coefficient of variation of cash calibration were less then 15%. A panel contained 8 serial dilutions was established as the national liner HBsAg reference panel. The permitted range of every dilution was stipulated and the stability of the panel was detected by accelerated test. Conclusions The national quantity standard of hepatitis B surface antigen was established and the national quantitative reference panel for HBsAg which contains eight liner serum was developed.