Aim To establish in vitro blood-brain barrier (BBB) model with characteristics of simulation of in vivo BBB by primi-tive co-culture of brain-microvessel endothelial cells (BMECs) with brain-microvessel pericytes (BMPC)and astrocytes (AS). Methods BMECs,BMPC and AS from SD rats were primitively isolated,purified and cultured,and then primitive culture cells were identified by cellular morphological and immunocytochemi-cal staining methods.Five types of in vitro BBB models were es-tablished by using Millicell culture insert (pore diameter 0.4μm)and their barrier functions were evaluated by detection of transendothelial electrical resistance (TEER),permeability of sodium fluorescent (Na-FLU ),expression of alkaline phospha-tase (AKP)and γ-glutamyl transpeptidase (γ-GT1 ),and simi-larity of permeation amount for positive drugs in vitro and in vivo BBB conditions.Results Primitive culture of BMECs presented typical pebbles-like structure,BMPC presented larger soma with branching property,AS presented slender synapse and shallower cytoplasm.Moreover,immunocytochemical staining results iden-tified primitive cells were targeted cells.TEER value for co-cul-ture of BMECs,BMPC and AS reached (478 ±25 )Ω· cm2 , permeability coefficients (Papp )value of Na-FLU was [(8.23 ± 0.78) ×10 -6 ]cm·s-1 ,expression of AKP and γ-GT1 were (6.90 ±0.27 )King unit · g-1 Pro and (4.39 ±0.32 )μg · g-1 Pro respectively.Moreover,good correlation could be found in Papp for positive controls in vitro and in vivo BBB models (R2=0.92).Conclusion The established in vitro BBB model by using primitive co-culture of BMECs with BMPC and AS posses-ses in vivo BBB properties in cell morphology,structures and barrier functions,and can be used as a powerful tool for studying physiology,pathology of BBB and screening candidate com-pounds.

Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.

Objective:To analyze the clinical data of 203 discharged patients with corona virus disease 2019(COVID-19), and to investigate the predictors for the severe cases.Methods:Confirmed COVID-19 cases hospitalized at Zhongnan Hospital of Wuhan University from January 1 to February 1, 2020 were consecutively enrolled, who were divided into severe group and non-severe group.The clinical data of enrolled patients were collected and the clinical manifestations, laboratory results, imaging, treatments and prognosis of patients in the two groups were analyzed. Mann-Whitney U rank sum test and chi-square test were used for statistical analysis. Results:A total of 203 discharged patients with COVID-19 were enrolled. The common clinical manifestations included fever (89.2%, 181/203), dry cough (60.1%, 122/203), chest distress (35.5%, 72/203), shortness of breath(29.1%, 59/203)and myalgia or arthralgia (26.6%, 54/203). The time from disease onset to hospital admission was 5.8 days (1.0 to 20.0 days). Among 203 enrolled patients, 107(52.7%) were divided into severe group and 96(47.3%) were non-severe group. The age in severe group was 60 years (23 to 91 years), which was significantly older than non-severe group (47 years (20 to 86 years)), the difference was statistically significant ( Z=-6.12, P<0.01). There were 63.6%(68/107) patients in severe group with at least one underlying disease, which was significantly more than non-severe group (20.8% (20/96)), the difference was statistically significant ( χ2=37.60, P<0.01). The proportions of patients with increased white blood cells, decreased lymphocytes and albumin, elevated alanine aminotransferase, aspartate aminotransferase, creatinine, lactic acid dehydrogenase, creatine kinase, fasting blood glucose, D-dimer, erythrocyte sedimentation rate, C-reactive protein, interleukin-6, and procalcitonin in severe group were all higher. On admission, 172 patients (84.7%) had bilateral patchy shadows or ground glass opacity in the lungs on chest imaging study, 20(9.9%) presented pleural effusion. Fifty-five cases (27.1%) showed progressions of lung lesions on computed tomography (CT) rescan at an average interval of five days. Among 203 patients, 123(60.6%) were given oxygen therapy upon admission, 107(52.7%) were given short-term glucocorticoid therapy, and 131(64.5%) received antiviral therapy; and 26(12.8%) died. The hospital stay was 11.0 days (1.0 to 45.0 days). Conclusions:Fever is the most common symptoms in COVID-19 patients.Elderly and patients with underlying diseases are risk factors for progression to severe cases. The elderly patients should be strengthened early monitoring, paid attention to the control of underlying diseases, and reduce the occurrence of critical diseases.

Objective:To investigate the effect of early enteral nutrition in patients with severe traumatic brain injury requiring exploratory abdominal surgery.Methods:The clinical data of 104 patients with severe traumatic brain injury requiring exploratory abdominal surgery treated in the PLA Army 72th Group Military Hospital from January 2016 to December 2020 were retrospectively analyzed. Among them, 47 patients were given early enteral nutrition (24-48 hr) as the observation group, and 57 patients were given delayed enteral nutrition (>48 hr), as the control group. The levels of hemoglobin, albumin, prealbumin, total bilirubin, alanine transaminase, C-reactive protein, white blood cells, postoperative infectious complications and clinical outcomes were compared between the two groups on the 1st, 7th and 14th days after surgery.Results:On the 14th day after operation, the prealbumin level of the observation group was higher than that of the control group, and the leukocyte level of the observation group was lower than that of the control group, with statistical significance ( P=0.020, P=0.013). The hospital stay and hospitalization costs of the observation group were lower than those of the control group ( P=0.017, P=0.032). The incidence of pulmonary infection in the observation group was 10.6%, which was lower than 29.8% in the control group ( P=0.017). Conclusions:Early postoperative enteral nutrition in patients with severe traumatic brain injury requiring exploratory abdominal surgery can significantly improve the nutritional status of patients, reduce the incidence of pulmonary infection, the hospital stay and the hospitalization cost.

Wnt proteins are a family of secreted glycoproteins that are evolutionarily conserved and considered to be involved in extensive developmental processes in metazoan organisms. The characterization of wnt genes may improve understanding the parasite's development. In the present study, a wnt4 gene encoding 491amino acids was amplified from cDNA of metacestodes of Taenia solium using reverse transcription PCR (RT-PCR). Bioinformatics tools were used for sequence analysis. The conserved domain of the wnt gene family was predicted. The expression profile of Wnt4 was investigated using real-time PCR. Wnt4 expression was found to be dramatically increased in scolex evaginated cysticerci when compared to invaginated cysticerci. In situ hybridization showed that wnt4 gene was distributed in the posterior end of the worm along the primary body axis in evaginated cysticerci. These findings indicated that wnt4 may take part in the process of cysticerci evagination and play a role in scolex/bladder development of cysticerci of T. solium.
Animals ; Base Sequence ; Cysticercosis/pathology ; Cysticercus/enzymology/*genetics ; DNA, Helminth/*genetics ; Gene Expression Regulation ; Humans ; In Situ Hybridization ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sus scrofa ; Swine ; Swine Diseases ; Taenia solium/embryology/enzymology/*genetics ; Wnt4 Protein/*genetics

Objective:To investigate the clinical features and prognosis of duodenal lateral fistula (DLF), and to explore the high-risk factors affecting its prognosis, so as to improve its treatment outcome.Methods:The regression study was conducted based on the database of the Registration System of Chinese Gastrointestinal Fistula and Intra-Abdominal Infections. DLF patients who were hospitalized from January 1, 2018 to December 31, 2018 and had complete clinical data in the database were selected as the research object. The clinical data included patient gender, age, length of hospital stay, hospitalization costs, intensive care unit (ICU) admission, underlying diseases, primary diseases, direct causes of DLF, complications, treatment and outcomes.Results:A total of 142 patients with DLF were enrolled, including 97 males and 45 females, with a median age of 54 years. The top three primary diseases of DLF were gastrointestinal ulcers and perforations in 33 cases (23.2%), biliary tract disease in 27 cases (19.0%), and trauma in 24 cases (16.9%). There were 117 cases (82.4%) with single DLF, and 25 cases (17.6%) were combined with other fistula. Among the 142 patients, 127 were healed (49 by surgery and 78 self-healing) and 15 (10.6%) died. 14 cases (9.9%) underwent percutaneous catheter drainage, and 18 cases underwent laparotomy drainage. Univariate prognostic analysis showed that advanced age (≥60 years old) (χ 2=6.891, P=0.009), primary diseases as gastrointestinal ulcers/perforations (χ 2=8.515, P=0.004), combined with other fistula (χ 2=5.798, P=0.016), malnutrition (χ 2=5.595, P=0.018), pulmonary infection (χ 2=12.449, P<0.001), hemorrhage (χ 2=6.466, P=0.011), multiple organ dysfunction syndrome(MODS ) (χ 2=37.258, P<0.001), underwent laparotomy drainage (χ 2=6.466, P=0.011) were associated with mortality. Multivariate prognostic analysis confirmed that age ≥60 years old ( OR=44.375, 95% CI: 2.676-735.822, P=0.008), combined with other fistula ( OR=16.54, 95% CI: 1.744-156.913, P=0.015) and MODS ( OR=238.447, 95% CI: 9.496-5 987.78, P=0.001) were independent risk factors of the death for DLF patients. Conclusions:The mortality rate of DLF is still relatively high. Iatrogenic duodenal injury (surgical operation, endoscopic treatment) is the main direct cause of fistula. Advanced age, combined with other fistula and MODS significantly increases the risk of death of patients.

Objective: Establish a murine model for hyperuricemia (HU) and periodontitis to explore whether there is correlation between them and provide a basis for periodontal treatment. Methods: Fourteen male KM mice were divided into 2 groups; the HU group (n=7) was fed food supplemented with potassium oxonate and uric acid, the NC group (n=7) was fed standard food, and the induction period was 35 days. On the 25th day, the molars on one side were ligated to induce periodontitis (P side), while the opposite was true for the control (C side). Baseline and terminal serum uric acid (UA) levels were detected, and alveolar bone resorption was analyzed by micro-CT. Results: The serum UA level of HU mice was (112.94 ± 26.82 )mol/L, that of the NC group was (72.21 ± 19.95) μmol/L, and the difference in UA level was statistically significant (P < 0.05). The P side bone volume fractions of the HU and NC groups were( 29.01 ± 11.09)% and (29.56 ± 15.27)%, respectively, which were not significantly different (t=-0.072, P=0.944). The P side bone mineral densities of the HU and NC groups were(0.53 ± 0.16) g/cm3 and (0.52 ± 0.14) g/cm3, respectively, which were not significantly different (t=0.038, P=0.970). Additionally, there was no correlation between HU or serum UA and alveolar bone resorption (P > 0.05). Conclusion This research established a murine model for HU and periodontitis, but based on micro-CT analysis of alveolar bone, no relationship between HU or UA levels and periodontitis was found.