Objective To investigate the constituents in Lysimachia congestiflora.Methods The constituents were isolated by the silica gel column chromatography,and their structures were identified by spectroscopy methods.Results ?-Stigmasterol(1),quercetin(2),myricetin 3′-methyl ether(3),syringetin-3-O-?-L-arabinofuranosid(4),ardicrenin(5) and ardisiacrispin A(6) were isolated and identified.Conclusion Six compounds have been isolated from Lysimachia congestiflora,and ardicrenin(5) is isolated from this species for the first time.

We established an ELISPOT for bovine interferon-gamma (BoIFN-γ), and applied it in the diagnosis of bovine tuberculosis (bTB). Monoclonal antibodies that can bind with native BoIFN-γ were screened as the coating antibody and detecting antibody. After optimization of detecting conditions including coating antibody concentration, cell number, and detecting antibody concentration, the ELISPOT assay was established. Peripheral mononuclear cells (PBMCs) isolated from 30 cows were co-cultured with PPD, and detected with the ELISPOT assay. The optimal conditions of ELISPOT assay were 2.5 μg/mL coating antibody 2G5, 2.5 x 10(5) cells/well, and 1 μg/mL detecting antibody Bio-5E11. In these 30 cows tested both with the ELISPOT assay and the BOVIGAM kit, 11 cows were proved to be positive in ELISOPT assay with the sensitivity of 78.6%, and 12 cows were proved to be negative in ELISOPT assay with the specificity of 75%. The ELISPOT assay for BoIFN-γ could be used to detect bTB efficiently and it might be an alternative method for the diagnosis of bTB.
Animals ; Antibodies, Monoclonal ; Cattle ; Enzyme-Linked Immunospot Assay ; veterinary ; Female ; Interferon-gamma ; isolation & purification ; Sensitivity and Specificity ; Tuberculosis, Bovine ; diagnosis

Manipulations are essential to the acupuncture curative effect. With the technique limited, the researches on acupuncture manipulations can only be carried out with respect to the senses and experience of doctors and patients, thus seriously impeding the modernization and internationalization of acupuncture. Both the modern integration technology of transducer and the biomechanical principles are applied to develop a detection system that can measure the interaction force between the manipulator and the manipulated patient on the needle at each kind of manipulation. Through clinical practice, the force waveforms acting on needle with the manipulations of symmetrical twirling-rotating and symmetrical lifting-inserting were recorded so as to realize the quantitative, objective and real-time detection of the force during the acupuncture process, which provided a new experimentation means and analysis method for the improvement of clinical curative effect and quantitative research of acupuncture and meridians.
Acupuncture ; instrumentation ; Biosensing Techniques ; instrumentation ; Equipment Design ; Humans ; Mechanics ; Medicine, Chinese Traditional ; Needles ; Software ; Software Design

OBJECTIVE: To investigate the effects of Naoluo Xintong Decoction (NLXTD) on pyroptosis and angiogenesis of brain microvascular endothelial cells (BMECs) and explore the possible mechanisms in rats with oxygen-glucose deprivation/ reperfusion (OGD/R). METHODS: Rat BMECs with or without caspase-1 siRNA transfection were cultured in the presence of 10% medicated serum from NLXTD-treated rats (or blank serum) and exposed to OGD/R. CCK-8 assay, Transwell chamber assay, and tube formation assay were used to assess proliferation, migration, and tube-forming abilities of the cells. The activity of lactate dehydrogenase (LDH) in the culture supernatant was determined using a commercial assay kit, and the levels of inflammatory factors IL-1β and IL-18 were detected with ELISA. The cellular expressions of pro-caspase-1, caspase-1, NLRP3, Gasdermin D, and angiogenesis-related proteins VEGF and VEGFR2 were detected using Western blotting. RESULTS: The BMECs showed obvious injuries after OGD/R exposure. Compared with the blank serum, the medicated serum significantly improved the cell viability, migration ability, and lumen-forming ability (P < 0.01) and lowered the levels of IL-1β and IL-18 and the LDH release (P < 0.01) of the cells with OGD/R exposure. Western blotting showed that in the BMECs exposed to OGD/R, the medicated serum strongly upregulated the expression of VEGF and VEGFR2 proteins (P < 0.01) and reduced the protein expressions of pro-caspase-1, caspase-1, NLRP3, and Gasdermin D (P < 0.01), and transfection of the cells with caspase-1 siRNA further promoted the expressions of VEGFR2 protein in the cells (P < 0.01). CONCLUSION NLXTD can improve the proliferation, migration, and tube- forming ability and promote angiogenesis of BMECs with OGD/R injury probably by inhibiting the caspase-1/Gasdermin D pathway in pyroptosis, alleviating cell injury, and upregulating the expressions of VEGF and VEGFR2.
Animals ; Rats ; Endothelial Cells ; Caspase 1 ; Gasdermins ; Interleukin-18 ; NLR Family, Pyrin Domain-Containing 3 Protein ; Vascular Endothelial Growth Factor A ; Reperfusion Injury ; Brain ; Angiogenic Proteins ; Glucose

Iro system and temperature-sensitive hemagglutinin (Tsh) genes were identified by suppression subtractive hybridization (SSH) and selective capture of transcribed sequences (SCOTS). To get more insights in the distribution and the occurrence of the iroC and tsh genes, we examined 243 avian E. coli strains for the presences of the these genes. Among 243 avian E. coli isolates, iroC gene was present in 84.4% strains (205/243). Of the 205 iroC-positive isolates, iroC gene was found in 184 (89.8%), 18(8.8%) and 3 (1.5%) isolates with high, intermediate and low pathogenicity, respectively. Of the 167 tsh-positive isolates, tsh gene was detected in 146 (87.4%), 21 (12.6%) and 0 (0%) isolates with high, intermediate and low pathogenicity, respectively. Among tsh-positive isolates, 89.5 to 100% of the highly pathogenic isolates of O1, O2 or O78 serogroups had the tsh gene, while 53.3% of the highly pathogenic isolates of non-O1, O2 and O78 serogroups had the tsh gene (P<0.01). Suicide vectors for deletion of the iroBCDEN or tsh genes were constructed as follows. The 715-bp fragments of iroB and 603-bp fragment of the iroN were generated by PCR respectively. Both of these two fragments together with EGFP gene were cloned into pUC18, termed pUC18-iroBNEGFP. A resultant suicide vector containing the iroB-EGFP-iroN fragment was obtained and named pMEG375-iroBNEGFP. Similarly, both of the 685-bp fragment of tshF and the 692-bp fragment of the tshR together with gentamycin gene were cloned into pUC18, resulting in pUC18-tshFRGm. A resultant suicide vector containing the tshF-Gm-tshR fragment was named pMEG375-tshFRGm. Mutant derivatives of strain E037 were generated by allelic replacements and were named E037(Deltairo), E037(Deltatsh) and E037(DeltairoDeltatsh). The 50% lethal dose (LD50) of E037, E037(Deltairo), E037(Deltatsh) and E037(DeltairoDeltatsh) in commercial day-old chickens experimentally inoculated via intratrachea were determined to be 10(5.6), 10(8.4), 10(9.0) and 10(9.5)CFU, respectively. In the chicken challenging model, the mutants were tested to determine the individual role of this system for virulence and persistence in chickens. The result suggested that Iro system and Tsh were important in the pathogenicity of APEC.
Adhesins, Escherichia coli ; genetics ; Animals ; Chickens ; Escherichia coli ; genetics ; pathogenicity ; Escherichia coli Infections ; microbiology ; veterinary ; Genes, Bacterial ; genetics ; Mutation ; Nucleic Acid Hybridization ; methods ; Organisms, Genetically Modified ; Poultry Diseases ; microbiology ; Transformation, Genetic ; Virulence Factors ; genetics

Objective To discuss our drug clinical trial institution's experience and findings during the process of establishing drug clinical trial central pharmacy.Methods Analyze the previous key issues identified during the drug management under different modes,discuss the necessity and feasibility of establishing drug clinical trial central pharmacy.Meanwhile,discuss the planning and construction of hardware including location site of the central pharmacy,equipment and facilities,staff,as well as software such as electronic management system and standard operation procedures.Results After the adoption of central trial pharmacy,space and energy are saved,manpower and material resources are saved,the quality of clinical trials also improved.Conclusions Standardized and unified management of investigational drugs through establishing drug clinical trial central pharmacy,is the strong guarantee for the drug safety of human subject,as well as the accuracy and scientificity of trial results.

Objective:To investigate the clinical efficacy of a medial support plate and a row of screws in the treatment of Schatzker type Ⅳ tibial plateau fracture involving the postero-lateral condyle.Methods:A retrospective analysis was performed of the data of 26 patients who had been admitted to Department of Orthopedic Trauma, Honghui Hospital Affiliated to Xi'an Jiaotong University for Schatzker type Ⅳ tibial plateau fracture involving the postero-lateral condyle from December 2015 to December 2020. There were 17 males and 9 females, aged from 19 to 51 years (average, 36.1 years). All their fracture lines involved the postero-lateral condyle and all fractures were fixated with a medial support plate and a row of screws via one medial and one lateral incisions. Recorded were the operation time, hospital stay, blood loss, incision length, fracture healing, complications, quality of knee joint reduction and knee joint function at the last follow-up.Results:The length of hospital stay ranged from 8 to 16 days, averaging 10.4 days. The 26 patients were followed up for 8 to 18 months, with an average of 14.3 months. All the fractures got united after 11 to 17 weeks (average, 13.7 weeks). During the follow-up, no reduction loss, internal fixation failure or surgical complications were observed. Rasmussen radiographic scores at the last follow-up ranged from 11 to 18 points, averaging 16.1 points. The range of motion of the knee joint ranged from 0° to 140°, averaging 120.8°. The Rasmussen scores of the knee function ranged from 14 to 28 points, averaging 24.4 points.Conclusion:In the treatment of Schatzker type Ⅳ tibial plateau fracture involving the postero-lateral condyle, the use of a medial support plate and a row of screws can achieve satisfactory clinical results.

Objective To evaluate the clinical application of a three-dimensional (3D) printing personalized guide for medial open wedge high tibial osteotomy(MOWHTO) in the treatment of knee varus osteoarthritis.Methods A retrospective study was conducted of the 16 patients with knee varus osteoarthritis who had been treated at Department of Orthopaedics,Zhengzhou Central Hospital of Zhengzhou University from January 2016 to January 2017.They were 6 men and 10 women,aged from 49 to 65 years (mean,55.8 years).Bilateral knees were involved in 2 cases and a unilateral knee was involved in 14 cases.Their disease duration ranged from one to 12 years (mean,5.3 years).A personalized guide for MOWHTO was designed and manufactured by 3D printing for every patient preoperatively.All the patients underwent knee arthroscopy before osteotomy which was assisted by the personalized guide.The femorotibial angle (FTA),medial proximal tibial angle (MPTA),weight bearing line (WBL),posterior tibial slope (PTSA) and the patellofemoral height Insall-Salvati index (IS index) were measured on their X-ray radiographs preoperatively and 6 months postoperatively.The Hospital for Special Surgery (HSS) score and visual analogue scale (VAS) were used to evaluate the outcomes at the final follow-ups.Results All surgeries were successful.The 16 patients were followed up for 6 to 12 months (mean,9.1 months).The FTA,MPTA,WBL and IS index at postoperative 6 months were significantly improved than the preoperative values (P ＜ 0.05).There was no significant difference between postoperative PTSA and preoperative PTSA (P =0.990).The mean VAS scores for the 16 patients at the final follow-ups were 0.8 ± 0.7 peints,significantly better than the preoperative ones (4.2 ± 0.9 points) (P ＜ 0.05);their mean postoperative HSS scores (89.3 ± 6.7 points) were also significantly improved than the preoperative ones (61.9 ± 10.5 points) (P ＜ 0.05).According to the HSS scores at the final follow-ups,the surgical outcomes were excellent in 14 knees,good in 3 and fair in one.Conclusion A 3D printed osteotomy guide can be used to perform precise osteotomy in MOWHTO for knee varus osteoarthritis,leading to effective correction of the alignment of the lower limb and good short-term surgical outcomes.

Rheumatoid arthritis (RA) is an autoimmune disease with angiogenesis, inflammatory factor infiltration and joint destruction as the main pathological features. Angiogenesis promotes the development of RA and plays an important role in its pathogenesis. The hypoxia-inducible factor (HIF)-vascular endothelial growth factor (VEGF)-angiopoietin-2 (Ang-2) signal transduction is a critical pathway to induce synovial angiogenesis. Targeting HIF-VEGF-Ang-2 signal transduction to inhibit synovial angiogenesis is a promising approach for RA treatment. This article reviews the role and mechanism of HIF-VEGF-Ang-2 signal transduction-mediated synovial angiogenesis in RA, in order to provide a new target and strategy for RA treatment.

Objective To establish high performance liquid chromatography(HPLC)characteristic chromatograms of Xinqingduyin Granules(composed of Taraxaci Herba,Lonicerae Japonicae Flos,Chrysanthemi Indici Flos,etc.)and content determination of chicory acid and glycyrrhizic acid,and to optimize the preparation process of Xinqingduyin Granules.Methods Using the characteristic chromatograms of Xinqingduyin and the retention rate of chicory acid and glycyrrhizic acid as indexes,we carried out orthogonal experiment to optimize the extraction process of Xinqingduyin,and studied the concentration process.The molding process of Xinqingduyin Granules was conducted by screening the types and dosage of auxiliary materials,then three batches of pilot experiments were carried out.Results HPLC characteristic chromatograms of Xinqingshuyin Granules and the determination methods of chicory acid and glycyrrhizic acid were established.The optimal preparation technology was as follows:8 times amount of water was added,the drug was decocted for 3 times,with 1 hour per time.After the extract was concentrated under reduced pressure at 80℃,the appropriate amount of steviol glycoside and lactose was added into the extract and mixed.One-step granulation and packaging were adopted.The retention rates of chicoric acid and glycyrrhizic acid in the 3 batches of Xinqingduyin Granules,which were prepared on the pilot scale,were(54.56±1.63)%and(54.96±1.08)%,and the rate of finished product was(87.47±0.49)%,respectively.The quality is uniform,and the characteristic map of Xinqingduyin Granules showed high similarity with that of decoction prepared from the same batch of slices.Conclusion The optimized preparation technology is reasonable,feasible and reproducible.This preparation can be used to obtain the granule with similar materials of Xinqingduyin decoction.