Objective To study the significance of DNA of Toxoplasma gondii in peripheral blood. Methods DNA of T.gondii in peripheral blood of 50 infected rats was detected by polymerase chain reaction. A pair of primers was designed, according to the sequence P30 gene specific to T.gondii, to amplify DNA from T.gondii by PCR. Results The primers amplified DNA specifically from T.gondii and could not amplify DNA from humans, uninfected rat and mouse and from Trichomonas vaginalis and Entamoeba histolytica. DNA of two Toxoplasma parasites was detected by 35 cycles of amplification, indicating a fair sensitivity of the PCR system. Conclusion PCR may have a value for early diagnosis of T.gondii infection in rat.

Time series model was developed to investigate the effect and contributions of related biomarkers on the cerebral endothelial dysfunction induced by beta amyloid(Aβ). HCMEC/D3 was incubated with 2. 5 μmol/L Aβ for 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 h, and biomarkers including cytosolic calcium ion, mitochondrial membrane potential(MMP), endothelial nitric oxide synthase(eNOS)and cell viability were determined. Time series model was established to assess the dynamic relationship between the related biomarkers and cell viability and the contribution of different biomarkers to cell damage. Impulse response analysis indicated that after a positive impact on cytosolic calcium ion, cell viability decreased and this impact continued to decline; after a positive impact on endothelial nitric oxide synthase and mitochondrial membrane potential, cell viability increases, which increased rapidly in the early stage, and the rate decreased in later stage. The result of variance decomposition showed that the cytosolic calcium ion played a major role in cerebral endothelial dysfunction induced by Aβ. Combined with the model study, it is concluded that the intervention on the level of cytosolic calcium ion at the early stage may be the possible way to slow the disease progression.

Objective:To develop a UPLC method for the simultaneous determination of 5 components in Guilong Kechuanning tablets including paeoniflorin, berberine hydrochloride, alkaloid, cinnamic acid and cinnamaldehyde. Methods:An ACQUITY UPLC BEH C18(2.1 mm ×100 mm,1.7 μm)chromatographic column was used;the mobile phase was acetonitrile(A)–0.1% formic acid solution (B) with gradient elution (0-10 min, 85% A;10-13 min, 10% A;13-15 min, 85% A) at a flow rate of 0. 4 ml·min-1, the detection wavelengths were:0-1. 8 min, 230 nm;1. 8-6. 0 min, 345 nm;6. 0-9. 0 min, 285 nm;9. 0-12. 0 min, 345 nm, and the column temperature was 30℃. Results:The linear range of paeoniflorin, berberine hydrochloride, alkaloid, cinnamic acid and cinna-maldehyde was 0. 060-1. 202 μg(r=0. 9999),0. 100-2. 010 μg(r=0. 9999),0. 040-0. 794 μg(r=0. 9994),0. 015-0. 302 μg(r=0.9999) and 0.042-0.850 μg(r =0.9999), the average recovery (n = 6) was 99.63%,99.26%,100.17%,98.80% and 100. 26%, and the RSDs were 0. 39%,0. 97%,0. 73%,1. 00% and 0. 71%, respectively. Conclusion:The method is simple, accu-rate and reproducible. It can be used for the quality control of Guilong Kechuanning tablets.

Objective:To develop a UPLC method for the simultaneous determination of 5 components in Guilong Kechuanning tablets including paeoniflorin, berberine hydrochloride, alkaloid, cinnamic acid and cinnamaldehyde. Methods:An ACQUITY UPLC BEH C18(2.1 mm ×100 mm,1.7 μm)chromatographic column was used;the mobile phase was acetonitrile(A)–0.1% formic acid solution (B) with gradient elution (0-10 min, 85% A;10-13 min, 10% A;13-15 min, 85% A) at a flow rate of 0. 4 ml·min-1, the detection wavelengths were:0-1. 8 min, 230 nm;1. 8-6. 0 min, 345 nm;6. 0-9. 0 min, 285 nm;9. 0-12. 0 min, 345 nm, and the column temperature was 30℃. Results:The linear range of paeoniflorin, berberine hydrochloride, alkaloid, cinnamic acid and cinna-maldehyde was 0. 060-1. 202 μg(r=0. 9999),0. 100-2. 010 μg(r=0. 9999),0. 040-0. 794 μg(r=0. 9994),0. 015-0. 302 μg(r=0.9999) and 0.042-0.850 μg(r =0.9999), the average recovery (n = 6) was 99.63%,99.26%,100.17%,98.80% and 100. 26%, and the RSDs were 0. 39%,0. 97%,0. 73%,1. 00% and 0. 71%, respectively. Conclusion:The method is simple, accu-rate and reproducible. It can be used for the quality control of Guilong Kechuanning tablets.

Objective:To explore the influencing factors of early postoperative complications after radical resection of congenital choledochal cyst (CCC) in a single center and provide some clinical basis and guidance for reducing postoperative complications.Methods:Case control study.Clinical data of 124 children (29 boys and 95 girls) with CCC diagnosed and radically treated at the Affiliated Hospital of Zunyi Medical University from September 2010 to October 2019 were analyzed.According to postoperative complications (bile leakage, gastrointestinal anastomotic fistula, bleeding, incision dehiscence, cholangitis, abdominal infection, pancreatitis, and lymphatic fistula), these children were divided into the complication group (group A) and non-complication group (group B). Age, laboratory indicators[preoperative white blood cell (WBC) count, hemoglobin, glutamic pyruvic transaminase, prealbumin, and postoperative albumin], and clinical factors, such as operation method, operation time, intraoperative blood loss, cyst type, cyst diameter, hepatic duct diameter, abdominal operation history, biliary sludge and calculus, hepatic duct anatomic variation, and pancreaticobiliary maljunction were statistically analyzed between the two groups.The t-test was performed for normal distribution of the measurement data, and the non-parametric rank sum test for non-normal distribution.Multivariate analysis was made using Logistic regression. Results:Among the 124 children, 25(20.16%) had complications, and 99(79.84%) had no complications.Bile leakage occurred in 14 children (11.29%), of whom 7 received operation again and 7 received conservative treatment.Gastrointestinal anastomotic fistula occurred in 2 children (1.61%), of whom 1 was re-operated and 1 was cured conservatively.One child (0.81%) was complicated with bleeding and cured by re-operation.Two children (1.61%) were complicated with incision dehiscence, of whom 1 was cured by re-operation and 1 was cured by conservative treatment.Cholangitis in 2 children (1.61%), abdominal infection in 2 children (1.61%), pancreatitis in 1 child (0.81%), and lymphatic fistula in 1 child (0.81%) were all conservatively cured.No significant difference was found in non-normal distribution indicators-age and WBC count-between the two groups (all P>0.05). Blood loss volume and cyst diameter were significantly different between the two groups (all P<0.05). Postoperative albumin[(27.84±4.62) g/L vs.(32.45±3.72) g/L] meeting the normal distribution showed a statistically significant difference between the two groups ( t=5.254, P<0.05). Logistic multivariate regression analysis suggested that preoperative anemia ( OR=7.922, 95% CI: 1.468-42.757) and biliary sludge and calculus ( OR=1.295, 95% CI: 1.075-4.359) were independent risk factors for postoperative complications; postoperative albumin ( OR=0.055, 95% CI: 0.012-0.244) was a protective factor for postoperative complications, and the differences were statistically significant (all P<0.05). Conclusions:The larger the cyst diameter, the more the intraoperative bleeding, and the higher the risk of operation.Treating anemia before operation, clearing sludge in the hepatic duct during operation, reducing bleeding, and strengthening the monitoring of albumin and hemoglobin during the perioperative period can prevent and reduce early complications after radical resection of CCC in children.

Objective:To prepare 68Ga-fibroblast activation protein inhibitor (FAPI)-04, and evaluate its biodistribution and imaging characteristics in animals and healthy volunteers, in order to investigate the clinical translation potential. Methods:68Ga-FAPI-04 was synthesized by a manual method and its radiolabeling yield, radiochemical purity, and stability ( in vivo and in vitro) were analyzed. ICR mice ( n=16) were scarified at 5, 30, 60 and 120 min postinjection of 68Ga-FAPI-04 (1.11 MBq) to measure radioactive counts in main organs. The dynamic mircoPET imaging was acquired for 60 min on 3 ICR mice, and tumor imaging capabilities were examined with nude mice bearing HepG2 tumors. Furthermore, 2 healthy volunteers (1 male with age of 64 years, 1 female with age of 56 years) were recruited for the investigation of probe biodistribution in humans. A serial whole-body dynamic PET/CT scan was performed immediately following injection. Results:68Ga-FAPI-04 was synthesized within 20 min with the radiochemical yield of (68.7±4.0)% (decay corrected). The radiochemical purities of 68Ga-FAPI-04 were over 99% and the products were stable for 180 min in vitro and for 90 min in blood. 68Ga-FAPI-04 was mainly cleared through urinary tracts, while other organs only showed mild tracer accumulation. MicroPET imaging showed high uptake of 68Ga-FAPI-04 in the tumor tissue of mice, and the ratio of tumor/liver was 2.14±0.01 (35 min). The PET/CT imaging results of healthy volunteers revealed 68Ga-FAPI-04 could be quickly cleared. Conclusion:68Ga-FAPI-04 has many advantages for PET imaging, such as easy labeling, good stability, quick clearance and low background signals in the liver, which can be used as an attractive PET tracer for detection hepatocellular carcinoma.

Objective:To investigate the radiation dosimetry and biodistribution of 68Ga-FAPI-04 PET/CT in patients with hepatobiliary tumor. Methods:A total of six patients with hepatic lesions who underwent PET/CT examination in Peking Union Medical College Hospital were enrolled. After intravenous injection of radiotracer 68Ga-FAPI-04 at (170.57 ± 14.43) MBq, whole-body imaging were performed at the time points of 3, 10, 15, 20, 30 and 60 min, respectively. Biodistribution pattern was observed. Regions of interest were manually delineated. Radiation dosimetry of all target organs were calculated by Olinda/EXM software. Results:The radioactive uptake dissipated gradually in liver whereas it was relatively stable in tumor lesions. The average SUV max of tumor lesions reached the maximum value (13.87± 2.55) at 20 min after injection. The target-to-background ratio increased with time, reaching the maximum value (10.09 ± 8.17) at 30 min after injection. The average effective dose in total body was (0.020 ± 0.002) mSv/MBq and organ with the highest effective dose was bladder wall at (0.146 ± 0.035) mSv/MBq. Conclusions:The effective dose in total body of 68Ga-FAPI-04 was similar to that of 18F-FDG. 68Ga-FAPI-04 is expected to be a PET/CT radiotracer for hepatobiliary tumors in consideration of rapid tumor uptake, low accumulation of liver background, and no influence of blood sugar levels.

Objective To establish in vitro the small intestinal organoid culture system and to investigate the effect of lipopolysaccharide(LPS)on the growth of small intestinal organoids and the secretion of inflammatory factors.Methods In vitro,the small intestinal crypt cell mass of C57BL/6 mice was aseptically isolated,collected and embedded in organoid matrix.Under the support of complete medium,the small intestinal organoids with three-dimensional multi-leaf structure with small intestinal epithelioid structure were formed.The small intestinal organoids were subcultured after 5-7 d culture.On the third day after passage,the small intestinal organoids were randomly divided into different mass concentrations of LPS groups(0,150,175,200,225,250,275 and 300 mg/L).After 24 h and 48 h of LPS induction,morphological changes of small intestinal organoid growth and differentiation were observed.CCK-8 method was used to detect the effect of different time points and mass concentrations of LPS on the proliferative activity of small intestinal organoids after induction of inflammation.The effects of four different mass concentrations of LPS(0,175,200 and 225 mg/L)on expression levels of granulocyte-macrophage colony stimulating factor(GM-CSF),interleukin(IL)-1α,IL-6 and IL-10 in organoid culture supernatant at different times were detected by enzyme-linked immunosorbent assay(ELISA).Results The mouse small intestinal organoid culture system was preliminarily constructed.After different time and mass concentration of LPS induced inflammation of small intestinal organoids,it was observed by morphology that small intestinal organoids would have different degrees of expansion and apoptosis in lumen.The proliferation,differentiation and budding of damaged intestinal epithelial crypts or intestinal stem cells were also inhibited to varying degrees,indicating that the growth of small intestinal organoids would be limited to varying degrees after induced inflammation.The proliferation activity of small intestinal organoids decreased to varying degrees after 24 h and 48 h of LPS induction at 175-225 mg/L(P＜0.05),but the cell viability was still greater than 50%.The levels of IL-1α,IL-6 and GM-CSF partially increased after induction with 200 mg/L and 225 mg/L LPS for 24 h and 48 h(P＜0.05).The level of IL-10 decreased after induction with 200 mg/L LPS for 24 h and 48 h(P＜0.05).Conclusion In this study,a model of intestinal inflammatory injury in vitro induced by LPS with different mass concentrations and time points is preliminarily constructed,which provides a more reliable research platform for the mechanism research of intestinal diseases and the screening of effective drugs in the future.

The lung is an important open organ and the primary site of respiration. Many life-threatening diseases develop in the lung, e.g., pneumonia, asthma, chronic obstructive pulmonary diseases (COPDs), pulmonary fibrosis, and lung cancer. In the lung, innate immunity serves as the frontline in both anti-irritant response and anti-tumor defense and is also critical for mucosal homeostasis; thus, it plays an important role in containing these pulmonary diseases. Innate lymphoid cells (ILCs), characterized by their strict tissue residence and distinct function in the mucosa, are attracting increased attention in innate immunity. Upon sensing the danger signals from damaged epithelium, ILCs activate, proliferate, and release numerous cytokines with specific local functions; they also participate in mucosal immune-surveillance, immune-regulation, and homeostasis. However, when their functions become uncontrolled, ILCs can enhance pathological states and induce diseases. In this review, we discuss the physiological and pathological functions of ILC subsets 1 to 3 in the lung, and how the pathogenic environment affects the function and plasticity of ILCs.
Animals ; Humans ; Immunity, Innate ; Lung ; immunology ; pathology ; Lung Diseases ; immunology ; pathology ; therapy ; Lymphocytes ; immunology ; pathology