The regulatory roles of Fasciola hepatica GSTO1 protein in inflammatory cytokine expression and apoptosis in murine macrophages.

Fascioliasis, a global zoonotic parasitic disease, is mainly caused by Fasciola hepatica (F. hepatica) parasitizing in the livers of hosts, mainly humans and herbivores. Glutathione S-transferase (GST) is one of the important excretory- secretory products (ESPs) from F. hepatica, however, the regulatory roles of its Omega subtype in the immunomodulatory effects remain unknown. Here, we expressed F. hepatica recombinant GSTO1 protein (rGSTO1) in Pichia pastoris and analyzed its antioxidant properties. Then, the interaction between F. hepatica rGSTO1 and RAW264.7 macrophages and its effects on inflammatory responses and cell apoptosis were further explored. The results revealed that GSTO1 of F. hepatica owned the potent ability to resist oxidative stress. F. hepatica rGSTO1 could interact with RAW264.7 macrophages and inhibit its cell viability, furthermore, it may suppress the production of pro-inflammatory cytokines IL-1ß, IL-6 and TNF-α, but promote the expression of anti-inflammatory cytokine IL-10. In addition, F. hepatica rGSTO1 may down-regulate the ratio of Bcl-2/Bax, and increase the expression of pro-apoptotic protein caspase-3, thereby eliciting the apoptosis of macrophages. Notably, F. hepatica rGSTO1 inhibited the activation of nuclear factor-κB (NF-κB) and mitogen­activated protein kinases (MAPKs p38, ERK and JNK) pathways in LPS-activated RAW264.7 cells, exerting potent modulatory effects on macrophages. These findings suggested that F. hepatica GSTO1 can modulate the host immune response, which provided new insights into the immune evasion mechanism of F. hepatica infection in host.

Molecular Characteristics and Genetic Diversity of Fasciola Hepatica from Sheep in Xinjiang, China.

Introduction: Fasciola hepatica is a trematode infecting ruminants worldwide and occasionally affecting other animal species, including humans. It causes significant economic losses. Geographic distribution and patterns of infection must be considered before control and management measures are developed for this parasite. DNA molecular markers are useful for the identification of flukes and elucidation of their genetic evolution. Therefore, the population structure of F. hepatica was studied using this method in sheep in Xinjiang, China. Material and Methods: The molecular characteristics, genetic relationships within the population and dispersal patterns of F. hepatica isolates were analysed based on the cox1 and nad1 genes. The population structure of F. hepatica from three regions of Xinjiang was explored and a neutrality test was conducted. Results: The cox1 and nad1 genes have 21 and 42 variable sites, respectively, which can be classified into 34 and 33 haplotypes. Median-joining network and phylogenetic tree analyses showed that there was no significant variation in F. hepatica isolates between the three geographical regions. Analysis of variance revealed that the genetic variation of F. hepatica was mainly present within the populations. The neutrality test indicated that the populations were relatively stable but the Hami population may have undergone short-term expansion. Conclusion: This study revealed for the first time the molecular characteristics, genetic diversity and dispersal patterns of F. hepatica isolates from sheep in Xinjiang, thus providing new insights into the genetic variation and haplotype diversity of F. hepatica from indigenous sheep.

Molecular Characteristics and Potent Immunomodulatory Activity of Fasciola hepatica Cystatin

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.

Occurrence of Gastrointestinal Parasites in Camels in the Tianshan Mountains Pastoral Area in China.

INTRODUCTION: Gastrointestinal parasites are some of the most common pathogens which are seriously harmful to the camel's health. The infection status of gastrointestinal parasites in camels (Camelus bactrianus) in the Tianshan Mountains pastoral area in China is still unclear. The aim of this study was to investigate the species and infection intensity of gastrointestinal tract parasites in local camels. MATERIAL AND METHODS: A total of 362 fresh faecal samples were collected and examined for parasite eggs using the saturated saline floating and natural sedimentation method. The parasite eggs were subjected to morphological and molecular examination and identification, and the infection rate and mean intensity of the parasites were analysed. RESULTS: A total of 15 gastrointestinal tract parasite species' eggs were identified, with a detection rate of 100%. Ostertagia spp. (100%) and Trichostrongylus spp. (98.1%) were dominant. Camels were often coinfected by 5-14 species. The average number of eggs per gram of faeces was higher for Ostertagia spp. (298), Haemonchus contortus (176) and Nematodirus spp. (138). The number of species of parasites infecting young camels was significantly lower than that of adult camels, but the infection intensity in young camels was significantly higher. CONCLUSION: Gastrointestinal parasites were highly prevalent in camels from the Tianshan Mountains pastoral area in China. This finding provides important epidemiological data for the prevention and control of associated infections in camels.

Effect of new simple breathing apparatus on oxygen therapy in patients with severe and critical coronavirus disease 2019 / 中华危重病急救医学

Objective:To make a new simple respirator and observe the oxygen therapy effect of the respirator on patients with severe and critical coronavirus disease 2019 (COVID-19).Methods:Based on the infectivity and hospital requirements of COVID-19, a new simple respirator was designed by the medical staff of the Department of Anesthesiology of the Second Affiliated Hospital of Nanchang University, which was applied on the 22 patients with severe and critical COVID-19 who needed oxygen therapy admitted to the Cancer Center of Tongji Medical College of Huazhong University of Science and Technology from February 15th to March 15th in 2020. The new simple respirator contained two National Utility Model Patents (a respirator: ZL 2015 2 0410623.6, a fluid switch and oxygen suction device: ZL 2017 2 0873509.6), which was mainly composed of anesthesia mask and filter, L-shaped connecting tube, soft breathing bladder, connecting tube and elastic fixing belt. When in use, the anesthesia mask was fixed to the patient's mouth and nose with elastic straps, the connecting tube was inserted into the oxygen meter interface, the oxygen flow was adjusted to 6-10 L/min, and the L-shaped connecting tube was opened immediately after the soft breathing bag was full. The carbon dioxide and excess oxygen in the body was discharged from exhaust port. The oxygen flow was lowered to 2-3 L/min, the patient's respiratory rate (RR) was observed through the soft breathing bag fluctuations, and the oxygen flow was adjusted at any time. The changes of pulse oxygen saturation (SpO 2), RR and heart rate (HR) before and after application of new simple respirator were observed, and the blood gas test results of part of the patients were collected. Results:Twenty-two patients with severe and critical COVID-19 had significantly higher SpO 2 at 10 minutes after application of the new simple ventilator than before application (0.994±0.007 vs. 0.952±0.017, P < 0.01), and RR was significantly lower than that before application (times/min: 27.59±3.63 vs. 29.64±3.81, P < 0.01); after 1 day of application, each index was further improved. All 13 patients who received blood gas analysis indicated no carbon dioxide accumulation. Conclusions:The new simple respirator can significantly improve the oxygen therapy effect of patients with severe and critical COVID-19. At the same time, 2019 novel coronavirus (2019-nCoV) can be filtered through the filter to reduce the formation of aerosol and protect the medical staff and patients.

Multi-Epitope Fusion Protein Eg mefAg-1 as a Serodiagnostic Candidate for Cystic Echinococcosis in Sheep.

Cystic echinococcosis (CE) in sheep is a hazardous zoonotic parasitic disease that is caused by Echinococcus granulosus (Eg). At present, serological test is an important diagnostic method for Eg infection in domestic animals. Here, a fusion protein Eg mefAg-1 harboring 8 dominant B-cell epitopes of Eg such as antigen B, tetraspanin 1, tetraspanin 6, reticulon and Eg95 was produced in E. coli and evaluated for CE in sheep by indirect ELISA. Eg mefAg-1 showed in ELISA a high sensitivity (93.41%) and specificity (99.31%), with a coincidence rate of 97.02%. Overall, it is suggested that the Eg mefAg-1 could be a potential antigen candidate for CE serodiagnosis in sheep.

Molecular Detection and Genetic Diversity of porcine Circovirus Type 3 in Commercial Pig Farms in Xinjiang Province, China.

INTRODUCTION: Porcine circovirus type 3 (PCV3) is a newly discovered porcine circovirus. The molecular characteristics and genetic evolution of PCV3 in Xinjiang province, China still being unclear, the aim of the study was their elucidation. MATERIAL AND METHODS: A total of 393 clinical samples were collected from pigs on commercial farms in nine different regions of Xinjiang and phylogenetic analysis based on full-length Cap genes was performed. RESULTS: The prevalence at farm level was 100%, while in all the tested samples it was 22.39%. Nine PCV3 strains were detected in Xinjiang province and they shared 98.9-99.3% nucleotide and 97.5-100.0% Cap gene amino acid sequence identities with other epidemic strains from China and abroad. Compared with other epidemic strains of PCV3, there were 26 base mutation sites in the Cap gene in the nine Xinjiang strains, resulting in the mutation of amino acids at positions 20, 24, 75, 77, 108, 111 and 206. Phylogenetic analysis showed that these strains can be divided into two different genetic groups, to the first of which five strains affiliated and divided between subgroups 1.1 and 1.2, and to the second of which the other four strains affiliated and similarly divided between subgroups 2.1 and 2.2. CONCLUSION: PCV3 circulates widely among commercial pig farms in Xinjiang province, China, and displays obvious genetic diversity. The results provide epidemiological information useful for the prevention and control of PCV3 infection in the pig industry.

Effect of preoperative sleep deprivation on hippocampal TLR4/NF-κB signaling pathway in aged mice with postoperative cognitive dysfunction / 中华麻醉学杂志

Objective To evaluate the effect of preoperative sleep deprivation on hippocampal Tolllike receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling pathway in aged mice with postoperative cognitive dysfunction.Methods Sixty clean-grade healthy male C57BL/6J mice,aged 16 months,weighing 28-36 g,were divided into 4 groups (n=15 each) using a random number table method:control group (group C),surgery group (group S),sleep deprivation group (group SD) and sleep deprivation plus surgery group (group SD+S).Mice were fed a common diet in group C.Mice were sleep-deprived for 24 h in group SD.Tibial fracture internal fixation was performed in group S.Tibial fracture internal fixation was performed after 24-h sleep deprivation in group SD+S.Cognitive function was assessed using the contextual fear conditioning test at days 3 and 7 after operation.The animals were sacrificed at day 7 after operation,brains were removed and hippocampi were isolated for determination of tumor necrosis factor-alpha (TNF-α) content (by enzyme-linked immunosorbent assay) and expression of TLR4 and p-NF-κB p65 (by Western blot).Results Compared with group C,the percentage of time spent freezing induced by condition and percentage of time spent freezing induced by context were significantly decreased at days 3 and 7 after operation,the content of TNF-α was increased,and the expression of TLR4 and p-NF-κB p65 was up-regulated in S,SD and SD+S groups (P＜0.05).Compared with group S and group SD,the percentage of time spent freezing induced by condition and percentage of time spent freezing induced by context were significantly decreased at days 3 and 7 after operation,the content of TNF-α was increased,and the expression of TLR4 and p-NF-κB p65 was up-regulated in group SD+S (P＜0.05).Conclusion Preoperative sleep deprivation further accentuates postoperative cognitive dysfunction,and the mechanism is related to activating TLR4/NF-κB signaling pathway and inducing inflammatory responses of aged mice.

Inhibition of the neutral sphingomyelinase-2 pathway protects against cerebral ischemia reperfusion injury in rats / 国际脑血管病杂志

Objective To investigate the roles of neutral sphingomyelinase-2 (nSMase2) pathway on cerebral edema and cerebral injury in cerebral ischemia-reperfusion injury in rats. Methods Seventy-six adult male SD rats were randomly divided into sham operation group (n = 12), modeling group (n = 16), vehicle group (n = 16), SB203580 (a p38 mitogen activated protein kinase [MAPK] inhibitor) treatment group (n = 16) , and MRS1754 (a selective adenosine A2B receptor [A2B AR] antagonist) treatment group (n = 16) according to the random number table. A suture-occluded method was used to induce a middle cerebral artery ischemia-reperfusion model. Vehicle, SB203580, and MRS1754 were injected into the lateral ventricles 30 min before model preparation, the neurological function score was performed after ischemia-reperfusion for 24 h. 2,3,5 triphenyltetrazolium staining was used to detect the infarct volume. The water content of brain tissue was detected by dry-wet weight method. Western blot analysis was used to detect the expression of nSMase 2 and p38 MAPK in ischemic brain tissue. Immunohistochemical staining was used to detect the expression of nSMase 2 in ischemic brain tissue. Results MRS1754 significantly decreased neurobehavioral score (P < 0. 05) and reduced cerebral infarction volume (P < 0. 05) in rats. Both MRS1754 and SB203580 significantly decreased the water content of ischemic brain tissue (all P < 0. 05). In addition, MRS1754 also significantly decrease the phosphorylation of p38 MAPK after ischemia-reperfusion and decreased the expression level of nSMase 2 (P < 0. 01). Conclusion Regulation of A2BAR and p38 MAPK of nSMase upstream may play a neuroprotective role after cerebral ischemia-reperfusion injury.

Role of fNIRS technology in observing the effect of needling Hegu (LI 4) on the functions of prefrontal cortex in healthy volunteers / 针灸推拿医学（英文版）

Objective: To observe the effect of needling Hegu (LI 4) on functions of the prefrontal cortex using the functional near infrared spectroscopy (fNIRS). Methods: A total of 12 healthy volunteers were randomly assigned to an acupuncture group (n=6) and a control group (n=6). The control group received no intervention. The acupuncture group received needling Hegu (LI 4) on the right side. The real-time concentration changes of oxygenated hemoglobin (HbO2), deoxygenated hemoglobin (d-Hb) and total hemoglobin (t-Hb) in the prefrontal cortex were recorded using the fNIRS, allowing for effect analysis of needling Hegu (LI 4) on the functions of the prefrontal cortex. Results: Compared with the control group, the HbO2 and t-Hb concentrations were increased in the acupuncture group and there were between-group statistical significances (bothP0.05). Conclusion: Needling Hegu (LI 4) can extensively activate the prefrontal cortex and therefore has significant effect on the functions of the brain's prefrontal cortex.

Study of coupled plasma filtration adsorption therapy for patients with severe infection and multiple organ dysfunction syndromes / 中华急诊医学杂志

Objective To investigate the effect of coupled plasma filtration adsorption (CPFA) on plasma cytokines:TNF-α,IL-1β,IL-6,cellular immunity,blood lactate acid concentration,heart rate,respiration rate,oxygenation index,hemodynamics,blood cells counts,and prognosis in patients with multiple organ dysfunction syndromes (MODS).Methods This was a prospective,randomized clinical trial in 45 patients diagnosed as MODS.Patients were randomly assigned to hemoperfution with resin adsorption (HP) + continuous venous-venous hemofiltration (CVVH) group,CPFA group and CVVH group.The general clinical data,APACHE Ⅱ score,number of failure organ and previous mentioned biomarkers were documented.Blood samples were collected before and after blood filtration with any one of these procedures.The plasma samples were isolated and stored with frozen at-60 ℃.Data were statistically analyzed with SPSS 13.0 version software.Results In CPFA group,plasma cytokines,TNF-α、IL-1β、IL-6,decreased markedly after plasma adsorption for two hours (P ＜ 0.01);and plasma concentrations of IL-6 were further descended after subsequent CVVH for 10 hours (P ＜ 0.05).In HP + CVVH group,plasma cytokines,TNF-α、IL-1β、IL-6,decreased markedly after HP (P ＜ 0.01),and plasma concentrations of IL-6 were further descended after subsequent CVVH for 10 hours (P ＜ 0.05).In CVVH group,plasma cytokines,TNF-α、IL-1β、IL-6,decreased after CVVH for 12 hours (P ＜ O.05).Blood lactate acid concentration,heart rate,respiration rate,oxygenation index,T-lymphocytes subgroups (CD3 +,CD4 +,CD8 +,CD4 +/CD8 + ratio),clinical symptoms were improved and dose of vasoactive agent was reduced in the patients of three groups without differences among them.The counts of red blood cells,white blood cells and platelets after CPFA and CVVH showed no significant changes.There was no significant difference in blood cell counts between CPFA and CVVH groups.After HP + CVVH,there was a trend of decrease in platelet count (P ＜ 0.05).Platelet counts were significanfly higher in patients treated with CPFA and CVVH group than those in patients treated with HP + CVVH group (P ＜ 0.05).There were 6 patients died in HP + CVVH group,6 patients died in CPFA group and 5 patients died in CVVH group within 28days.Conclusions The comparison of efficacy of blood filtration among 3 modalities of HP + CVVH,CPFA and CVVH showed CPFA had higher capacity of Inflammatory medium scavenging than CVVH,and had less damage effect on blood visible component,especially on platelet compared with HP + CVVH.CPFA was an effective and safety modality in the treatment of the patients with multiple organ dysfunction syndrome.

Effect of exercise preconditioning on nerve growth factor and learning-and-memory abilities in rats suffered from focal cerebral ischemia / 中华行为医学与脑科学杂志

Objective To investigate the effect of exercise preconditioning on the expression of nerve growth factor(NGF) and its receptor TrkA as well as learning-and-memory abilities in rats suffered from focal cerebral ischemia.Methods Thirty-six male SD rats were randomly divided into sham operation group (Sham-MCAO,n=12),focal cerebral ischemia-reperfusion group (MCAO,n=12) and exercise preconditioning + cerebral ischemia-reperfusion group (EX+MCAO,n=12).Rats in EX+MCAO group were placed in the treadmill device and accepted 4 weeks exercise training.Then method of middle cerebral artery occlusion was applied to prepare transient focal cerebral ischemia reperfusion model.Garcia's method was used to assess the neural function in rats.Western blotting was applied to test the expression of NGF and TrkA protein in the successfully established experimental MCAO rats.Morris water maze experiment was uti lized to test the learning-and-memory abilities of the rats.Results (1) Compared with Sham-MCAO group,the expression of NGF in rats' brain tissue was lower in MCAO group (cerebral ischemia 1h reperfusion 24h) (P＜0.05).The expression of NGF of EX+MCAO group was higher than that of MCAO group,but still lower than that of Sham-MCAO group (P＜0.05).(2)The expression of TrkA in rats' brain tissue was higher in MCAO group compared with Sham-MCAO group (P＜0.05).Compared with MCAO group,the expression of TrkA was even higher in EX+MCAO group (P＜0.05).(3)On the fifth day in the Morris water maze test,the latency of MCAO group was significantly longer than that of Sham-MCAO group((9.36± 1.18)s vs (4.86± 1.52) s,P＜0.05).However,compared with MCAO group,the latency in EX+MCAO group ((6.02± 1.04) s) was shorter,but still longer than Sham-MCAO group (P＜0.05).There was no significant difference among the three groups in the average swimming speed (P＞0.05).Conclusion Exercise preconditioning can up-regulate the expressions of NGF and TrkA protein,which can also improve the learning-and-memory abilities in rats suffered from focal cerebral ischemia reperfusion injury.

Determination of taurine in biological samples by high-performance liquid chromatography using 4-fluoro-7-nitrobenzofurazan as a derivatizing agent / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>A highly sensitive and rapid high-performance liquid chromatography method with pre-column derivatization with 4-fluoro-7-nitrobenzofurazan was developed for determination of taurine in biological samples, including plasma, brain, and liver.</p><p><b>METHODS</b>The optimum derivatization reaction temperature was 70 °C, and at this temperature the reaction was complete within 3 min. The derivatized taurine was separated using phosphate buffer (0.02 mol/L, pH 6.0):acetonitrile (84:16, v/v) as the mobile phase at a flow rate of 1.0 mL/min, and a column temperature of 25 °C. The taurine derivatives were separated within 20 min (tR:14.5 min) and fluorometrically detected at 530 nm with excitation at 470 nm.</p><p><b>RESULTS</b>The intra- and the inter-day coefficients of variation for the method were 5.3% and 7.7%, respectively. The calibration curve was linear from 0.1 μmol/L to 30.0 μmol/L with a correlation coefficient of 0.9995.</p><p><b>CONCLUSION</b>This method can be used to determine the taurine contents in plasma, brain, and liver from normal rats and human plasma.</p>

Clinical investigation of mild cognitive impairment in patients with Parkinson's disease / 中华神经科杂志

Objective To describe the prevalence and neuropsychological character of mild cognitive impairment (MCI) associated with Parkinson' s disease(PD-MCI). Methods One hundred and three PD patients and a control group of 32 healthy old subjects were chosen. Psychometric assessment included the Mini Mental State Examination, the Dementia Rating Scale and a series of neuropsychol ogicaltests. The Hamilton Rating Scale of Depression was used to assess depression in PD patients. Results (1)Twenty-one (20.4%) PD patients was diagnosed with dementia, 45 (43.7%) had a MCI and only 37(35.9%) had no cognitive impairment; (2) Subjects with PD-MCI were older, had a later onset of the PD,and displayed more severe motor symptoms compared with those without cognitive impairment; (3) The prevalence and neuropsychological profile of PD-MCI were thought to correlate with the dominating side and subtype of Parkinsonian symptoms, for patients with left-sided dominant symptoms had a significantly higher chance of suffering MCI than those with right-sided dominant symptoms, the ratio being 74.2% vs 42.2%,χ<'2 =7. 589,P <0.05; The tremor-dominant group took less time than the mixed group for Stroop word test measurement ((80.8±39.9) s vs (94.4±30.0) s,t=3.332,P<0.01). Conclusion Identification of MCI is of important clinical significance, which helps to treat patients differently and thus predict the prognosis.

The effects of medium pH and purification method on product rate of expression rhGM-CSF in recombinant E.coli / 中国生化药物杂志

Purpose　The aim is to study the influence factors of recombinant E.coli cell growth,product expression and purification,in order to raise the product rate of rhGM-CSF.Methods　Three pH media were planed to observe the effects of pH on E.coli cell growth and expression with the indexes of the cell density and expression level.The influences of various treatments on product rate were observed with the indexes of the volume and purity of inclusion bodies and the yield,purity,activity of product.Using SDS-PAGE method for analysis of purity.The product bioactivity was determined by using TF1 cell and MTT colorimetric method.Results　The cell density in the medium pH 7.0 was 2.22 g/L;the product expression rate was 23%.It was two times higher than other pH media.The inclusion body prepared by bacteria without freezing store was 2.5 times volume than freezing store.During renatura-tion PEG 4000 could raise 0.8 times active product.The optimum conditions used rhGM-CSF product rate could raise 8 times.Concluston　The product rate of rhGM-CSF expressed in E.coli was affected by many factors.The optimum condition for culture,expression and purification could raise quite a number of product rate.