Object To identify the differences of chemical constituents of Dragon's Blood obtained by different extract processing. Methods The mentioned Dragon's Blood was analyzed by TLC, UV spectro photometry, and HPLC. Results The contents of dihydrochalcone in Dragon's Blood by the new extract processing were higher than that by traditional one's, while the contents of flavonoid and stibene were lower. Conclusion The heating free technology is a new technique that is an original with higher extract rate, and it is worth of spreading. It will promote the modernization of TCM.

A method was developed for the determination of zeranols (α,β-zearalanol,α,β-zearalenol,zear-alanone,zearalenone) with RRLC-MS/MS in the plant tissue. The samples were extracted with acetonitrile and reextracted with aqueous alkaline solution,cleaned up with MAX column and then determined by RRLC-MS/MS using multiple reaction monitoring ( MRM) scan mode. The results showed that the working curves were linear in the range of 0 -20 μg/kg. The limits of detection ( LOD) of zeranols were 0.5 μg/kg and the limits of quantification (LOQ) was 1. 0μg/kg. Extraction recoveries for zeranols ranged from 75. 8% to 105.4% and RSD was between 2.4% and 12.1%. The method is suitable for the determination of zeranols in the plant tissue.

Vaccination is considered a promising alternative for controlling tick infestations. Haemaphysalis longicornis midgut proteins separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membrane were screened for protective value against bites. The western blot demonstrated the immunogenicity of 92 kDa protein (P92). The analysis of the P92 amino acid sequence by LC-MS/MS indicated that it was a H. longicornis paramyosin (Hl-Pmy). The full lenghth cDNA of Hl-Pmy was obtained by rapid amplification of cDNA ends (RACE) which consisted of 2,783 bp with a 161 bp 3' untranslated region. Sequence alignment of tick paramyosin (Pmy) showed that Hl-Pmy shared a high level of conservation among ticks. Comparison with the protective epitope sequence of other invertebrate Pmy, it was calculated that the protective epitope of Hl-Pmy was a peptide (LEEAEGSSETVVEMNKKRDTE) named LEE, which was close to the N-terminal of Hl-Pmy protein. The secondary structure analysis suggested that LEE had non-helical segments within an alpha-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.
Amino Acid Sequence ; Animals ; Antigens/genetics/*metabolism ; Base Sequence ; Blotting, Western ; Chromatography, Liquid ; *Cloning, Molecular ; DNA, Complementary/genetics ; Epitopes ; Gene Expression Regulation/*physiology ; Ixodidae/genetics/*metabolism ; Molecular Sequence Data ; Tandem Mass Spectrometry

In order to explore tick proteins as potential targets for further developing vaccine against ticks, the total proteins of unfed female Dermacentor silvarum were screened with anti-D. silvarum serum produced from rabbits. The results of western blot showed that 3 antigenic proteins of about 100, 68, and 52 kDa were detected by polyclonal antibodies, which means that they probably have immunogenicity. Then, unfed female tick proteins were separated by 12% SDS-PAGE, and target proteins (100, 68, and 52 kDa) were cut and analyzed by LC-MS/MS, respectively. The comparative results of peptide sequences showed that they might be vitellogenin (Vg), heat shock protein 60 (Hsp60), and fructose-1, 6-bisphosphate aldolase (FBA), respectively. These data will lay the foundation for the further validation of antigenic proteins to prevent infestation and diseases transmitted by D. silvarum.
Animals ; Antigens/*chemistry/immunology ; Arthropod Proteins/*chemistry/immunology ; Electrophoresis, Polyacrylamide Gel ; Female ; Ixodidae/*chemistry/immunology ; Molecular Weight ; Rabbits ; Tandem Mass Spectrometry

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.
Animals ; Antigens/chemistry/genetics/*immunology/isolation & purification ; Arthropod Proteins/chemistry/genetics/*immunology/isolation & purification ; Chromatography, Affinity ; Cloning, Molecular ; Cluster Analysis ; Conserved Sequence ; Dermacentor/*genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli/genetics ; Gene Expression ; Humans ; Molecular Sequence Data ; Molecular Weight ; Phylogeny ; Recombinant Proteins/chemistry/genetics/immunology/isolation & purification ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

Objective:To investigate the clinicopathological features, immunohistochemical phenotypes and molecular characteristics of adenosquamous carcinoma of the lung(ASC)in elderly patients.Methods:Clinical data of 72 ASC patients in the Department of Pathology, The Second Affiliated Hospital of Soochow University from January 2009 to December 2020 were retrospectively analyzed, and 48 patients aged ≥60 years were selected.Clinical manifestations, imaging findings, histopathological and immunohistochemical characteristics were collected, and gene mutations were detected by the amplification refractory mutation system(ARMS-PCR).Results:There were 48 patients including 32 males and 16 females with a mean age of 70 years(range: 60-84 years). The maximum diameters of the tumors ranged from 0.3 to 9.0 cm(mean: 2.8 cm). Microscopically, the tumors contained two components, squamous cell carcinoma and adenocarcinoma, with the squamous cell carcinoma tissue showing intercellular bridges and the adenocarcinoma tissue showing papillary, acinar or tubular structures.Immunohistochemistry assays detected varying expression levels of CK7(30/31), CK5/6(20/28), TTF1(12/31), P40(15/17), and P63(12/13). Molecular testing showed that the EGFR mutation rate was 58.8%(10/17)and the ALK fusion mutation rate was 5.9%(1/17), while ROS1 and MET mutations were not detected.All 48 patients underwent surgical resection.Conclusions:ASC cases are relatively rare and prone to misdiagnosis.The diagnosis requires the combination of HE morphology, immunohistochemistry and imaging examination, and surgery is the main treatment option.The mutation rate of the EGFR gene is relatively high in ASC patients.

OBJECTIVE：To shar e the experienc e of pharmaceutical care in Wuhan Jinyintan Hospital （herein after refers to “our hospital ”）under the condition of novel coronavirus pneumonia （COVID-19）epidemic，and to provide reference for other hospitals to deal with public health emergencies. METHODS ：The situation of pharmaceutical care in our hospital under the condition of COVID- 19 epidemic was summarized and shared ，including the epidemic prevention and control management （regional division ，disinfection management ，pharmacy personnel training ），supply of drugs and disinfection products ，the monitoring and education of rational drug use by information technology. RESULTS ：The pharmacy department of our hospital divided the activity scope into clean area ，potential pollution risk area ，semi pollution area ，and implement different disinfection management. All pharmacists received training ，involving personal health protection ，prevention and control knowledge of COVID-19，health status monitoring ，etc. For supply and guarantee of drugs and disinfectants ，the epidemic drug list of our hospital was formulated ，drugs and disinfectants were purchased accurately and stored in a standardized way. 24 h response telephone was set up in the clinical pharmacy room to receive consultation from clinicians on drug use at any time. The drugs mentioned in the COVID- 19 diagnosis scheme were compared in terms of the mechanism of action and the medication of special populations to form a tablet ，so as to help clinical rational choice treatment drug. CONCLUSIONS ：The pharmaceutical care in the designated hospital of COVID- 19 is a professional and complicated work ，involving a wide range of aspects. Pharmacy department must respond actively and adjust the strategy in time so as to play an important role in improving the ability of medical treatment.

Chronic atrophic gastritis (CAG), a common disease of digestive system, is an extremely important cause of gastric cancer (GC). The occurrence and development of CAG involves the abnormality of multiple signaling pathways. Traditional Chinese medicines (TCMs) has the advantages of mild action, multi-target and small adverse reaction, etc., which broadens the way for the treatment of the disease, and TCMs can play a therapeutic role by regulating multiple signaling pathways. In this review, based on the related experiments of TCMs and Chinese herbal compounds in recent years, the related literatures were searched and 10 kinds of signaling pathways involved were summarized, in order to provide a reference for further research on reversing or delaying the progress of CAG and preventing gastric cancer.

The genus Rosa (Rosaceae family) includes about 200 species spread in the world, and this genus shows unique advantages in medicine and food. To date, several scholars concentrated on compounds belonging to flavonoids, triterpenes, tannins, polysaccharide, phenolic acids, fatty acids, organic acids, carotenoids, and vitamins. Pharmacological effects such as antineoplastic and anti-cancer properties, anti-inflammatory, antioxidant, liver protection, regulate blood sugar, antimicrobial activity, antiviral activity, as well as nervous system protection and cardiovascular protection were wildly reported. This article reviews the chemical constituents, pharmacological effects, applications and safety evaluations of Rosa plants, which provides a reference for the comprehensive utilization of medicine and food resources and gives a scientific basis for the development of medicinal plants of the genus Rosa.

OBJECTIVE To investigate the inhibitory effect of berberine （BER） on the invasion and migration of human renal carcinoma cells and its potential mechanism. METHODS Using human renal carcinoma OSRC-2 cell as object， alamarBlue assay was adopted to detect the inhibitory effects of 0 （control group）， 25， 50， 75， 100， 125， 150， 175 and 200 μmol/L BER on the proliferation of OSRC-2 cell after treatment for 24 h and 48 h. After treated with 0（control group）， 50， 100 μmol/L BER for 48 h， the effect of BER on cell cycle was analyzed by flow cytometry. The migration of OSRC-2 cells in 24 h and 36 h was observed by cell scratch test， and the invasion ability of OSRC-2 cells in 24 h was detected by Transwell assay. The protein expression of methyltransferase-like 3 （METTL3） was detected by Western blot after treatment for 48 h， and RNA methylation quantification kit was used to detect the levels of N6-methyladenosine （m6A） in OSRC-2 cells. RESULTS Compared with control group， BER at different concentrations could significantly decrease the survival rate of OSRC-2 cells （P＜0.01）， and showed a dose-dependent and time-dependent manner. After 48 h of BER treatment at 50， 100 μmol/L， the cell was arrested in G0/G1 phase （P＜0.01）. Compared with control group， the migration and invasion abilities of cells in 50， 100 μmol/L BER group were significantly decreased （P＜0.05 or P＜0.01）； the protein expression of METTL3 and the level of m6A in RNA were significantly decreased （P＜0.01）. CONCLUSIONS BER can inhibit level of m6A by down-regulating the expression of METTL3， thereby inhibiting the invasion and metastasis of human renal carcinoma cells.