Liver injury in COVID-19: Holds ferritinophagy-mediated ferroptosis accountable.

Even in patients without a history of liver disease, liver injury caused by coronavirus disease 2019 (COVID-19) is gradually becoming more common. However, the precise pathophysiological mechanisms behind COVID-19's liver pathogenicity are still not fully understood. We hypothesize that inflammation may become worse by cytokine storms caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Elevated ferritin levels can initiate ferritinophagy mediated by nuclear receptor coactivator 4 (NCOA4), which leads to iron elevation, and ferroptosis. In COVID-19 patients, ferroptosis can be restricted to reduce disease severity and liver damage by targeting NCOA4-mediated ferritinophagy. To confirm the role of ferritinophagy-mediated ferroptosis in SARS-CoV-2 infection, further research is required.

Liberal versus restrictive fluid management in abdominal surgery: a meta-analysis.

PURPOSE: This study compared perioperative restrictive fluid therapy to liberal (conventional) fluid therapy in patients undergoing major abdominal surgery and investigated the rate of post-operative morbidity (complication rates), recovery (time to flatus), and the length of hospital stay. METHODS: The Medline, PubMed, Cochrane, and EMBASE databases were searched until June 18, 2015. Randomized controlled trials, two-arm prospective studies, and retrospective studies were included in our analyses. A sensitivity analysis, publication bias assessment, and quality assessment were performed. RESULTS: The effects of the two therapies were similar in the subgroup analysis of patients who underwent hepato-gastroenterological surgery (P = 0.287). However, in a subgroup of patients who underwent vascular abdominal surgery, the restricted fluid treatment regimen was associated with a lower risk of complications in comparison with the conventional regimen (pooled OR = 0.12, 95 % CI 0.03-0.47, P = 0.002). There was no difference between the two regimens with respect to the incidence of cardiopulmonary complications (P = 0.733). However, the patients who received the restricted fluid treatment regimen had a shorter time to flatus (P = 0.031) and a shorter hospital stay (P = 0.033) than the patients who received the conventional regimen. CONCLUSIONS: Restrictive fluid therapy and liberal conventional therapy were associated with similar rates of overall and cardiopulmonary complications; however, restrictive fluid therapy was associated with a more rapid recovery and a shorter length of hospital stay.

Establishment of lymphatic filarial specific IgG4 indirect ELISA detection method.

OBJECTIVE: To establish the lymphatic filarial specific IgG4 indirect ELISA detection method and develop the kits. METHODS: ELISA and the developed specific IgG4 reagent was used to explore the best way for detecting filarial specific IgG4. Combined with the production process of commercialized enzyme immunoassay kit to develop economical lymphatic filarial specific IgG4 test kit, and to explore the value of the kit in the laboratory. RESULTS: We determined the most optimal detective antigen was Malay adult filarial antigen and the optimal concentration of coating antigen was 1.0 µg/ml. The appropriate serum dilution was 1:20 to 40 and the work titers of specific IgG4 agents was 1:800. We determined the optimal reaction time for substrates and developed a reproducible and stable detection kit with sensitive and specificity, which was easy to operate. CONCLUSION: We successfully established the lymphatic filarial specific IgG4 indirect ELISA detection method and developed the kits with good reproducibility and stable result, which should be widely applied.

Construction and identification of Complex DNA vaccine of hepatitis B and Toxoplasma gondii.

OBJECTIVE: To construct and identify multi-gene recombinant expression vector pcDNA3-HBsAg-p30-ROP2. METHOD: Primers were designed according to the gene sequences of restriction enzyme cutting site of recombinant pcDNA3-p30-ROP2 and hepatitis B surface antigen (HBsAg). The target fragment of HBsAg was amplified and cloned to expression vector pcDNA3-p30-ROP2 by restriction enzyme digestion and ligation. The recombinant expression vector pcDNA3-HBsAg-p30-ROP2 was identified by PCR detection, followed by enzyme restriction and sequencing. RESULTS: The target fragment of HBsAg was successfully amplified, and the multi-gene eukaryotic expression vector pcDNA3-HBsAg-p30-ROP2 was established. PCR detection and restriction enzyme digestion showed that the length of the target fragment was consistent with the theoretical value. The recombinant expression vector contained the complete sequences of p30-ROP2 compound gene and HBsAg. CONCLUSION: Multi-gene recombinant expression vector pcDNA3-HBsAg-p30-ROP2 was successfully established. The constructed expression vector could be used to develop multi-gene nucleic acid vaccines.

[Cloning, expression, purification and identification of Toxoplasma gondii SAG2 gene in Escherichia coli].

OBJECTIVE: To construct a recombinant plasmid containing surface antigen 2(SAG2) gene of Toxoplasma gondii and express it in Escherichia coli. METHODS: The truncated SAG2 gene was amplified from the genomic DNA of T. gondii RH strain and cloned into plasmid pGEX-4T. Then the recombinant pGEX-4T-SAG2 was induced by IPTG and expressed in E. richia col BL21. The expressed proteins were analyzed by SDS-PAGE and purified, and the immunogenicity of the product was analyzed by Western blotting. RESULTS: The amplified SAG2 gene was about 561 bp, which was accorded to the expectation. The recombinant plasmid was constructed successfully by digested with double restriction enzyme and confirmed with DNA sequencing. SDS-PAGE and Western blotting showed the molecular weight of SAG2 fusion protein was about 47 ku, and the protein could be identified by GST-tag antibody. CONCLUSION: The truncated SAG2 gene of T. gondii has been successfully cloned and expressed in E. coli BL21 cells, and the recombinant protein has immunogenicity.

[Extraction Method of Malaria Parasite DNA from Preserved Positive Blood Smears].

OBJECTIVE: To develop a method for DNA extraction from malaria parasites on preserved blood smears, to provide basis for research on malaria genetic traceability. METHODS: The improved DNA extraction kit (QIAamp DNA Mini Kit) was used to extract plasmodium DNA from 41 giemsa-stained blood smears, and the extraction was compared with that using the Chelex-100 and Na(2)HPO(4) methods. Nested PCR was used to amplify small subunit ribosomal RNA to identify Plasmodium parasite. The PCR products underwent sequencing and sequence alignment, to analyze the difference in PCR positive rates between blood smears prepared in the 1980s and in recent 10 years, between blood smears with and without deoil/decoloration, and between blood smears with different qualities. RESULTS: The total PCR positive rate for the improved kit method was 70.7% (29/41). The PCR positive rate for blood smears prepared in the 1980s and in recent 10 years was 78.6% (11/14) and 66.7% (18/27) respectively, with no significant difference (W=0.63, P>0.05). The PCR positive rate for blood smears with and with- out deoil/decoloration was 62.5% (15/24) and 82.4% (14/17) respectively, also with no significant difference (χ(2)= 1.89, P>0.05). However, the PCR positive rate was significantly higher in blood smears with high quality [93.3% (28/30)] than those with low quality [9.1%(1/1l)](=27.59, P<0.01). Sequence alignment showed that the PCR products were consistent with the target DNA fragments. However, DNA extracted using the Chelex-100 and Na(2)HPO(4) methods showed negative PCR results. CONCLUSIONS: DNA extracted from blood smears prepared in the 1980s using the improved Kit (QIAamp DNA Mini Kit) shows a high PCR positive rate. Besides, blood smear staining and use of oil for microscopic examination do not affect DNA extraction.

Protection of MES23.5 dopaminergic cells by obestatin is mediated by proliferative rather than anti-apoptotic action.

Obestatin is an endogenous peptide sharing a precursor with ghrelin. This study aims to investigate whether and how obestatin protects MES23.5 dopaminergic cells against 1-methyl-4-phenylpyridinium (MPP(+))-induced neurotoxicity. MES23.5 cells were pretreated with obestatin (10(-13)-10(-6) mol/L) for 20 min prior to incubation with 200 µmol/L MPP(+) for 12 or 24 h, or treated with obestatin alone (10(-13) to 10(-6) mol/L) for 0, 6, 12, and 24 h. The methyl thiazolyl tetrazolium (MTT) assay was used to measure cell viability. Flow cytometry was used to measure the caspase-3 activity and the mitochondrial transmembrane potential. Proliferating cell nuclear antigen (PCNA) protein levels were determined by Western blotting. Obestatin (10(-13) to 10(-7) mol/L) pretreatment blocked or even reversed the MPP(+)-induced reduction of viability in MES23.5 cells, but had no effect on MPP(+)-induced mitochondrial transmembrane potential collapse and caspase-3 activation. When applied alone, obestatin increased viability. Elevated PCNA levels occurred with 10(-7), 10(-9), 10(-11) and 10(-13) mol/L obestatin treatment for 12 h. The results suggest that the protective effects of obestatin against MPP(+) in MES23.5 cells are due to its proliferation-promoting rather than anti-apoptotic effects.

Methods of patient warming during abdominal surgery.

BACKGROUND: Keeping abdominal surgery patients warm is common and warming methods are needed in power outages during natural disasters. We aimed to evaluate the efficacy of low-cost, low-power warming methods for maintaining normothermia in abdominal surgery patients. METHODS: Patients (n = 160) scheduled for elective abdominal surgery were included in this prospective clinical study. Five warming methods were applied: heated blood transfusion/fluid infusion vs. unheated; wrapping patients vs. not wrapping; applying moist dressings, heated or not; surgical field rinse heated or not; and applying heating blankets or not. Patients' nasopharyngeal and rectal temperatures were recorded to evaluate warming efficacy. Significant differences were found in mean temperatures of warmed patients compared to those not warmed. RESULTS: When we compared temperatures of abdominal surgery patient groups receiving three specific warming methods with temperatures of control groups not receiving these methods, significant differences were revealed in temperatures maintained during the surgeries between the warmed groups and controls. DISCUSSION: The value of maintaining normothermia in patients undergoing abdominal surgery under general anesthesia is accepted. Three effective economical and practically applicable warming methods are combined body wrapping and heating blanket; combined body wrapping, heated moist dressings, and heating blanket; combined body wrapping, heated moist dressings, and warmed surgical rinse fluid, with or without heating blanket. These methods are practically applicable when low-cost method is indeed needed.