In order to establish a simple,sensitive and specific diagnostic method for the simultane-ous detection of Mycoplasma hyorhinis(Mhr)and Mycoplasma hyopneumoniae(Mhp)in swine,specific primers and probes were designed using the Mhr p37 and Mhp p36 gene sequences as the target genes,and the dual LFD-RPA rapid test was established by screening the primers and probes,optimizing primer ratios and evaluating its effectiveness through the sensitivity,reproduc-ibility and clinical sample testing.The sensitivity,specificity,reproducibility and clinical samples were evaluated.The results showed that the established dual LFD-RPA assay could complete the amplification in 15 min at 39 ℃,and its optimal primer ratio was 1.6∶0.8,and the lowest detection limits were up to 3.63 and 3.60 copies/μL,respectively;the reproducibility was stable;and there was no cross-reactivity with Pasteurella multocida,Bordetella bronchiseptica,Haemophilus pa-rasuis,Actinobacillus pleuropneumoniae,Escherichia coli.The test successfully established a dual LFD-RPA assay,which can detect Mhr and Mhp simultaneously,and is simple,sensitive and spe-cific without relying on specialized equipment,and is suitable for carrying out on-site rapid diagno-sis of Mhr and Mhp.

In order to establish a simple,sensitive and specific diagnostic method for the simultane-ous detection of Mycoplasma hyorhinis(Mhr)and Mycoplasma hyopneumoniae(Mhp)in swine,specific primers and probes were designed using the Mhr p37 and Mhp p36 gene sequences as the target genes,and the dual LFD-RPA rapid test was established by screening the primers and probes,optimizing primer ratios and evaluating its effectiveness through the sensitivity,reproduc-ibility and clinical sample testing.The sensitivity,specificity,reproducibility and clinical samples were evaluated.The results showed that the established dual LFD-RPA assay could complete the amplification in 15 min at 39 ℃,and its optimal primer ratio was 1.6∶0.8,and the lowest detection limits were up to 3.63 and 3.60 copies/μL,respectively;the reproducibility was stable;and there was no cross-reactivity with Pasteurella multocida,Bordetella bronchiseptica,Haemophilus pa-rasuis,Actinobacillus pleuropneumoniae,Escherichia coli.The test successfully established a dual LFD-RPA assay,which can detect Mhr and Mhp simultaneously,and is simple,sensitive and spe-cific without relying on specialized equipment,and is suitable for carrying out on-site rapid diagno-sis of Mhr and Mhp.

This study aims to compare the prevalence of sexually transmitted infections (STIs) with semen quality in men from couples with primary and secondary infertility. Semen samples were collected from 133 men who requested fertility evaluation. Seminal tract infection with Ureaplasma spp. (UU), Mycoplasma hominis (MH), Mycoplasma genitalium (MG), Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and herpes simplex virus-2 (HSV-2) was assessed by PCR-based diagnostic assays. Among all patients, the prevalence of STIs was higher in men from couples with primary infertility than that in men from couples with secondary infertility (39.7% vs 21.7%, P = 0.03). The prevalence of UU was 28.8% and 13.3% in men from couples with primary and secondary infertility, respectively. Men from couples with primary infertility were more likely to be positive for UU than men from couples with secondary infertility (P = 0.04). Regarding the UU subtype, the prevalence of Ureaplasma urealyticum (Uuu) and Ureaplasma parvum (Uup; including Uup1, Uup3, Uup6, and Uup14) did not differ between the two groups. No associations between the prevalence rates of MH, MG, and CT were found in men from either infertility group. A lower sperm concentration was associated with STI pathogen positivity in men with primary infertility according to the crude model (P = 0.04). The crude and adjusted models showed that semen volume (both P = 0.03) and semen leukocyte count (both P = 0.02) were independently associated with secondary infertility. These findings suggest the importance of classifying the type of infertility during routine diagnosis of seminal tract infections.
Female ; Humans ; Infertility, Male/epidemiology* ; Male ; Mycoplasma genitalium ; Mycoplasma hominis ; Prevalence ; Semen ; Semen Analysis ; Sexually Transmitted Diseases/epidemiology* ; Ureaplasma urealyticum

The mixing process is one of the key operation units for solid preparation of traditional Chinese medicine. The physical properties such as particle size, density and viscosity of the mixture are key factors that need to be controlled, which will directly affect the performance of the preparation molding process and product quality. Subsequent dripping process performance and appearance qua-lity of dripping pills will be affected by dynamic viscosity of materials in the mixing process. Based on this, with mixing process of compound Danshen dripping pills as the object, a feedforward control method for the dripping pill mixing process was established based on the concept of quality by design(QbD). Firstly, critical quality attribute(CQA)-dynamic viscosity, critical material attributes(CMAs)-the moisture content of compound Danshen extract, average molecular weight of polyethylene glycol 6000 and critical process parameter(CPP)-mixing temperature were identified through the analysis of properties for multiple batches of the raw materials and excipients as well as technological mechanism. Then the Box-Behnken experimental design was used to establish the regression model among CMA, CPP and CMA(R■=0.972 0, RMSE =16.24) to obtain the design space. Finally, through the verification of three batches within the design space, the mixing process temperature was adjusted according to the properties of the raw materials and exci-pients to achieve accurate control of the dynamic viscosity attribute. The relative deviation between the actual dynamic viscosity value and the target value was less than 3.0 %. The feedforward control of the mixing process of compound Danshen dripping pills was rea-lized in this study, which can contribute to improving quality consistency of the mixing process intermediates, simultaneously provide a reference for the research on the process quality control of other Chinese medicine dripping pills.
Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Quality Control ; Research Design

An ultra-high performance liquid chromatography method for the determination of 8 constituents in Qingzao Jiufei Decoction was established and the basis of related chemical substances with antioxidant activity in Qingzao Jiufei Decoction was explored. The separation was performed on a Waters Cortecs RP Shield C18 (150 mm × 2.1 mm, 1.6 μm) using UHPLC-DAD as the mobile phase was water (containing 0.1% phosphoric acid) – acetonitrile with flow rate of 0.30 mL·min^-1 by gradient elution ① determining 5 constituents (amygdalin, liquiritin, liquiritin apioside, rutin and isoquercitrin) at the wavelength of 210 nm, 237 nm and 358 nm. Under gradient elution ②, 3 constituents (glycyrrhizin, glycyrrhizic acid and sesamin) were determined at the wavelength of 210 nm and 265 nm. The IC₅₀ of 10 batches of Qingzao Jiufei Decoction scavenging 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS⁺) free radicals obtained through test and Probit model was analyzed for correlation with the contents of 8 constituents. The established methods had a good linear relationship (r > 0.999), good repeatability and stability. The recovery rate was between 82.8% and 112.4%. In a series of concentration range, the higher the concentration of Qingzao Jiufei Decoction, the stronger the free radical scavenging effect. There was a significant correlation between the content of rutin and glycyrrhizic acid and the IC₅₀ of scavenging free radicals. The content determination methods established in this experiment provide a basis for a reasonable and scientific evaluation of the quality of Qingzao Jiufei Decoction. Qingzao Jiufei Decoction has antioxidant activity, which is significantly positively correlated with the content of rutin and glycyrrhizic acid.

A quantitative analytical method for multi-components with a single-marker (QAMS) was established for simultaneous determination of neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction. The separation was performed on a Waters CORTECS T3 column (2.1 mm × 100 mm, 2.7 μm), with the mobile phase consisting of 0.05% phosphate acid solution-acetonitrile for gradient elution. The column temperature was 30 ℃, and flow rate was 0.5 mL·min^-1. Using chlorogenic acid as an internal reference, the relative correlation factors of neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid were calculated following UHPLC, as 0.928 0, 0.546 2, 1.099 8, 0.872 1, 1.086 8, 0.739 2, 1.056 6, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method. There was no significant difference in assay results between QAMS and the external standard method. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the content such as neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction.

PURPOSETo investigate the effects of salvianolic acid B (SAB) on tumor necrosis factor a (TNF-α) induced alterations of cerebral microcirculation with a bone-abrading model.

METHODSThe influences of craniotomy model and bone-abrading model on cerebral microcirculation were compared. The bone-abrading method was used to detect the effects of intracerebroventricular application of 40 μg/kg·bw TNF-α on cerebral venular leakage of fluorescein isothiocyanate (FITC)- albulmin and the rolling and adhesion of leukocytes on venules with fluorescence tracer rhodamine 6G. The therapeutical effects of SAB on TNF-α induced microcirculatory alteration were observed, with continuous intravenous injection of 5 mg/kg·h SAB starting at 20 min before or 20 min after TNF-α administration, respectively. The expressions of CD11b/CD18 and CD62L in leukocytes were measured with flow cytometry. Immunohistochemical staining was also used to detect E-selectin and ICAM-1 expression in endothelial cells.

RESULTSCompared with craniotomy method, the bone-abrading method preserved a higher erythrocyte velocity in cerebral venules and more opening capillaries. TNF-α intervention only caused responses of vascular hyperpermeability and leukocyte rolling on venular walls, without leukocyte adhesion and other hemodynamic changes. Pre- or post-SAB treatment attenuated those responses and suppressed the enhanced expressions of CD11b/CD18 and CD62L in leukocytes and E-selectin and ICAM-1 in endothelial cells induced by TNF-α.

CONCLUSIONSThe pre- and post-applications of SAB during TNF-α stimulation could suppress adhesive molecular expression and subsequently attenuate the increase of cerebral vascular permeability and leukocyte rolling.

Animals ; Benzofurans ; pharmacology ; Blood Flow Velocity ; Cerebrovascular Circulation ; drug effects ; Craniotomy ; Disease Models, Animal ; E-Selectin ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Mice, Inbred C57BL ; Microcirculation ; drug effects ; Random Allocation ; Reference Values ; Tumor Necrosis Factor-alpha ; administration & dosage

OBJECTIVE: To explore the application value of multi-slice spiral computed tomography (MSCT) in traffic accidents through observing and analyzing the injury features of the accidents. METHODS: Two fatal cases caused by traffic accidents were fully examined using MSCT, 3D imaging reconstruction and angiography through cardiac puncture. The features of traffic injury mechanism were analyzed through combination of MSCT and postmortem external examination. RESULTS: In case 1, right cardiac rupture was found by MSCT and angiography through cardiac puncture. The cause of death was cardiac tamponade and right ventricular rupture due to the crush injury of chest in the traffic accident. In case 2, splenic rupture and intra-abdominal hemorrhage was found and caused by injury of left trunk by MSCT. The cause of death was hemorrhage and traumatic shock. CONCLUSION MSCT could observe skeletal injury, soft tissue injury, and hematologic disorder well. The combination use of MSCT and angiography through cardiac puncture provided assistance to the diagnosis of cardiovascular system injury.
Accidents, Traffic/mortality* ; Autopsy ; Fatal Outcome ; Hemorrhage ; Humans ; Imaging, Three-Dimensional/methods* ; Multidetector Computed Tomography/methods* ; Soft Tissue Injuries ; Wounds and Injuries/pathology*

Postmortem chemistry is becoming more and more essential in routine forensic pathology and has made considerable progress over the past years. Biochemical analyses of vitreous humor, blood, urine and cerebrospinal fluid may provide important information in determining the cause of death or in elucidating forensic issues. Postmortem chemistry may be essential for the determination of cause of death when morphological methods (diabetes mellitus, alcoholic ketoacidosis and electrolytic disorders) cannot detect the pathophysiological changes involved in the death process. It can also provide many information in other forensic situations, including myocardial ischemia, sepsis, inflammation, infection, anaphylaxis and hormonal disturbances. The most recent relevant research advances on glucose metabolism, liver function, cardiac function, renal function, sepsis, inflammation, infection, anaphylaxis and hormonal aspect are hereby reviewed.
Anaphylaxis ; Autopsy/trends* ; Biomarkers/analysis* ; Body Fluids/chemistry* ; Death ; Diabetes Mellitus ; Forensic Pathology/methods* ; Humans ; Postmortem Changes ; Sepsis ; Vitreous Body

The combination use of dexamethasone and calcium gluconate can be applied to hypersensitivity. Severe hypokalemia is a usual complication of dexamethasone and calcium gluconate therapy, which occurs frequently with therapeutic use. Fatal cases, accidental and intentional, occur frequently in forensic practice. The current case report presented a 43-year-old man with diabetes mellitus with infection, to whom dexamethasone and calcium gluconate were administered in the private clinic. With the development of such clinical symptoms of severe hypokalemia as quadriplegia, he was confirmed to have severe hypokalemia through a biochemical test before dying of arrhythmia. And also it presented pathophysiologic mechanism underlying severe hypokalemia as well as suggestions for clinical practice regarding combination use of dexamethasone and calcium gluconate.
Adult ; Anti-Inflammatory Agents/adverse effects* ; Calcium Gluconate/adverse effects* ; Dexamethasone/adverse effects* ; Diabetes Mellitus ; Fatal Outcome ; Humans ; Hypokalemia/chemically induced* ; Male