Identification of a natural PLA2 inhibitor from the marine fungus Aspergillus sp. c1 for MAFLD treatment that suppressed lipotoxicity by inhibiting the IRE-1α/XBP-1s axis and JNK signaling

Lipotoxicity is a pivotal factor that initiates and exacerbates liver injury and is involved in the development of metabolic-associated fatty liver disease (MAFLD). However, there are few reported lipotoxicity inhibitors. Here, we identified a natural anti-lipotoxicity candidate, HN-001, from the marine fungus Aspergillus sp. C1. HN-001 dose- and time- dependently reversed palmitic acid (PA)-induced hepatocyte death. This protection was associated with IRE-1α-mediated XBP-1 splicing inhibition, which resulted in suppression of XBP-1s nuclear translocation and transcriptional regulation. Knockdown of XBP-1s attenuated lipotoxicity, but no additional ameliorative effect of HN-001 on lipotoxicity was observed in XBP-1s knockdown hepatocytes. Notably, the ER stress and lipotoxicity amelioration was associated with PLA2. Both HN-001 and the PLA2 inhibitor MAFP inhibited PLA2 activity, reduced lysophosphatidylcholine (LPC) level, subsequently ameliorated lipotoxicity. In contrast, overexpression of PLA2 caused exacerbation of lipotoxicity and weakened the anti-lipotoxic effects of HN-001. Additionally, HN-001 treatment suppressed the downstream pro-apoptotic JNK pathway. In vivo, chronic administration of HN-001 (i.p.) in mice alleviated all manifestations of MAFLD, including hepatic steatosis, liver injury, inflammation, and fibrogenesis. These effects were correlated with PLA2/IRE-1α/XBP-1s axis and JNK signaling suppression. These data indicate that HN-001 has therapeutic potential for MAFLD because it suppresses lipotoxicity, and provide a natural structural basis for developing anti-MAFLD candidates.

Modern Research Progress of Polyphyllin Ⅰ： A Review / 中国实验方剂学杂志

Paridis Rhizoma， a traditional valuable Chinese herbal medicine， has the functions of clearing heat and removing toxin， relieving edema and pain， cooling liver and calming convulsion， which can be used to treat various diseases such as mumps， abscess， burn， bleeding， and tumor. It has been used in folk medicine for a long time and is the main raw material of various Chinese patent medicines such as Gongxuening Capsules and Yunnan Baiyao. Polyphyllin Ⅰ， an isospirostanol saponin and one of the main active components in Paridis Rhizoma， is distributed in the rhizome， pericarp， and leaves of Paris polyphylla. With high polarity， polyphyllin Ⅰ is mainly extracted by n-butanol extraction and macroporous adsorption resin chromatography， separated by silica gel column chromatography and preparative high performance liquid chromatography， and purified with the combination of methods. With anti-tumor， anti-inflammatory， antibacterial， and anti-virus effects， it is generally employed to treat liver cancer， lung cancer， gastric cancer and other cancers as well as arthritis， influenza， sore toxin， and bacterial infection. However， polyphyllin Ⅰ may cause stomach irritation， hemolysis， liver damage， kidney damage， heart damage， and other adverse reactions. Pharmacokinetic studies show that it has problems such as low bioavailability and poor intestinal absorption and permeability， which affect the clinical application of polyphyllin Ⅰ. This paper summarizes the research on the plant sources， extraction and separation methods， pharmacological effects， adverse reactions， and pharmacokinetics of polyphyllin Ⅰ in recent years， which is expected to provide a reference for the rational clinical application and other in-depth research work of polyphyllin Ⅰ.

Correlation analysis of serum TTR, RBP4 expressions and gestational metabolic syndrome / 中华内分泌外科杂志

Objective:To investigate the relationship between the expressions of serum trans thyroxine protein (TTR) , retinol binding protein 4 (RBP4) and metabolic syndrome of pregnancy (GMS) .Methods:A total of 103 patients with GMS from May. 2017 to Dec. 2019 were selected as the observation group, in addition, 100 healthy pregnant women with single pregnancy and no complications were selected as control group. Before pregnancy, the height and weight were detected. After the diagnosis of GMS, serum markers and blood pressure were detected. The levels of total cholesterol (TC) , triglyceride (TG) , low density lipoprotein cholesterol (LDL-C) , high density lipoprotein cholesterol (HDL-C) and fasting blood glucose (FBG) were detected by Hitachi 7600 automatic biochemical analyzer, and the expression levels of TTR and RBP4 in serum were detected by enzyme-linked immunosorbent assay (ELISA) . Receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic value of TTR and RBP4 in GMS. Multivariate Logistic regression model was used to analyze the influencing factors of GMS in pregnant women.Results:There was no significant difference in age or gestational weeks between the two groups ( P>0.05) . Compared with those in the control group, the levels of BMI, TC, TG, LDL-C, FBG, systolic blood pressure, diastolic blood pressure, serum TTR and RBP4 expression levels were higher in the observation group before pregnancy ( P<0.05) , while HDL-C was lower ( P<0.05) . The area under the curve (AUC) of TTR and RBP4 alone or combination in the diagnosis of GMS was 0.797, 0.816 and 0.898, respectively. The cut-off value of TTR was 284.91 mg/L, and the sensitivity and specificity were 63.10% and 89.00%, respectively. The cut-off value of RBP4 was 17.89 mg/L, and the sensitivity and specificity were 69.90% and 87.00%, respectively. The sensitivity and specificity of the combined diagnosis were 83.50% and 84.00% respectively. Multivariate Logistic regression analysis showed that high level of TTR and high level of RBP4 were independent risk factors of GMS in pregnant women ( P<0.05) . Conclusions:The expression levels of serum TTR and RBP4 in GMS patients are high, and they have certain diagnostic value for GMS. They are independent risk factors of GMS in pregnant women, which may provide new ideas for the early diagnosis and prevention of GMS.

Sudden sensorineural hearing loss and ischemic stroke / 国际脑血管病杂志

Sudden sensorineural hearing loss (SSNHL) refers to the sudden and unexplained sensorineural hearing loss within 72 h and a decrease in hearing of ≥30 dB affecting at least 3 consecutive frequencies. It is one of the common emergencies in neurology and otolaryngology. Early etiological evaluation and systematic and targeted treatment are very important for delaying the progression of SSNHL and restoring hearing. Recent studies have shown that SSNHL overlaps with vascular risk factors of ischemic stroke, and may predict the risk of ischemic stroke. SSNHL may be one of the clinical manifestation and even the prodromal symptoms of ischemic stroke, especially the infarction of the blood supply area of the anterior inferior cerebellar artery or its branch internal auditory artery. Although these factors can not fully reveal the relationship between SSNHL and ischemic stroke, they are enough to warn clinicians that they should consider the possibility of ischemic stroke when receiving patients with SSNHL. Screening of vascular risk factors for patients with SSNHL as early as possible is helpful to avoid the risk of recurrence of ischemic stroke.

Risk factors of non-obstructive gastric retention before endoscopic treatment in patients with biliopancreatic disease: a matched case-control study / 中华消化内镜杂志

Objective:To investigate the risk factors of non-obstructive gastric retention before endoscopic retrograde cholangiopancreatography (ERCP) in patients with biliopancreatic disease.Methods:The matched case-control study included 109 patients with non-obstructive gastric retention before ERCP as the case group, and 218 patients without gastric retention as the control group. The patients′ medical records including comorbidities, preoperative medication and laboratory indicators were compared between the two groups. Multivariate conditional logistic regression models were subsequently used to determine the risk factors for non-obstructive gastric retention before ERCP.Results:Logistic regression revealed that the jaundice ( OR=12.359, P<0.001), opiates use ( OR=3.009, P=0.001), somatostatin use ( OR=2.445, P=0.033), fasting hyperglycemia ( OR=1.513, P=0.045), hypokalemia ( OR=4.634, P=0.001) and hyponatremia ( OR=1.805, P=0.023) were independent risk factors for non-obstructive gastric retention before ERCP in patients with biliopancreatic disease. Conclusion:Except for gastrointestinal obstruction, jaundice, opiates use, somatostatin use, fasting hyperglycemia, hypokalemia and hyponatremia are all risk factors for gastric retention in patients with biliopancreatic disease. Comprehensive evaluation and early intervention for patients showing these risk factors are needed.

Electrical stimulation combined with manual massage can relieve myofascial pelvic pain / 中华物理医学与康复杂志

Objective:To evaluate the clinical effectiveness of combining electrical stimulation with manual massage in treating women′s myofascial pelvic pain (MPPS).Methods:A total of 93 MPPS patients were recruited and randomly divided into an infrared irradiation group ( n=30), an electrical stimulation group ( n=31) and a combination group ( n=32). Those in the infrared group and the electrical stimulation group were given 30 minutes of infrared irradiation or electrical stimulation daily for 10 days, while the combined group was given a manual massage with electrical stimulation. The response to treatment was evaluated using a visual analogue scale (VAS) to rate discomfort, plus a physical examination, and surface electromyography results using Glazer′s protocol. The treatments′ efficacy and the recurrence of pain were evaluated 1 month after the treatment. Results:The average VAS ratings, resting potentials and their variability of the three groups all improved significantly after the treatment. The average VAS scores of the combined and electrical stimulation groups after the treatment were significantly lower than that of the infrared group. Moreover, the total effective rates of the combined group (96.87%) and the electrical stimulation group (80.65%) were significantly higher than that of the infrared group (56.67%), and the resting potential and variability of the former two groups were also significantly better. All of the above measurements among the combined group after the treatment were significantly better, on average, than in the electrical stimulation group. A month later the recurrence rate in the combined group (6.25%) was significantly lower than in the electrical stimulation group (25.81%) and the infrared group (56.67%).Conclusion:Electrical stimulation combined with manual massage is effective in relieving myofascial pelvic pain.

Electrical stimulation combined with manual massage can relieve myofascial pelvic pain / 中华物理医学与康复杂志

Objective@#To evaluate the clinical effectiveness of combining electrical stimulation with manual massage in treating women′s myofascial pelvic pain (MPPS).@*Methods@#A total of 93 MPPS patients were recruited and randomly divided into an infrared irradiation group (n=30), an electrical stimulation group (n=31) and a combination group (n=32). Those in the infrared group and the electrical stimulation group were given 30 minutes of infrared irradiation or electrical stimulation daily for 10 days, while the combined group was given a manual massage with electrical stimulation. The response to treatment was evaluated using a visual analogue scale (VAS) to rate discomfort, plus a physical examination, and surface electromyography results using Glazer′s protocol. The treatments′ efficacy and the recurrence of pain were evaluated 1 month after the treatment.@*Results@#The average VAS ratings, resting potentials and their variability of the three groups all improved significantly after the treatment. The average VAS scores of the combined and electrical stimulation groups after the treatment were significantly lower than that of the infrared group. Moreover, the total effective rates of the combined group (96.87%) and the electrical stimulation group (80.65%) were significantly higher than that of the infrared group (56.67%), and the resting potential and variability of the former two groups were also significantly better. All of the above measurements among the combined group after the treatment were significantly better, on average, than in the electrical stimulation group. A month later the recurrence rate in the combined group (6.25%) was significantly lower than in the electrical stimulation group (25.81%) and the infrared group (56.67%).@*Conclusion@#Electrical stimulation combined with manual massage is effective in relieving myofascial pelvic pain.

Clinical significances of additional chromosome abnormalities and t (15;17) in acute promyelocytic leukemia / 白血病·淋巴瘤

Objective To investigate the clinical significances of additional chromosome abnormalities and t(15;17) in acute promyelocytic leukemia (APL). Methods A total of 90 newly diagnosed APL patients in the Affiliated Hospital of Qingdao University from January 2007 to June 2014 were analyzed retrospectively. Patients with different chromosome karyotypes were divided into four groups: additional chromosome number abnormalities group (16 cases), additional chromosome structural abnormalities group (14 cases), additional chromosome number and structural abnormalities group (4 cases) and typical chromosome group (56 cases). According to whether the patient contained t(15;17), the patients were divided into group with t (15;17) and group without t (15;17). The short-term efficacy and survival of each group were analyzed and compared. Results The rate of complete remission in additional chromosome number abnormalities group, additional chromosome structural abnormalities group, additional chromosome number and structural abnormalities group and typical t(15;17) chromosome changes group were 56.3％(9/16), 100.0％(14/14), 25.0％(1/4) and 82.1％(46/56), the early mortality rates were 25.0％(4/16), 0 (0/14), 75.0％(3/4) and 8.9％ (5/56) respectively. Among them, the additional number and structural abnormalities group had lower complete remission rate and higher early mortality rate, and compared with other groups, the differences were statistically significant (all P< 0.05). The complete remission rates of the group with t (15;17) and the group without t (15;17) were 80.5％ (66/82) and 50.0％ (4/8), respectively, and the difference was not statistically significant (P= 0.070). Conclusions APL patients with karyotypes with additional number and structural changes have low complete remission rate, high early mortality rate and poor prognosis. Patients with t(15;17)have a high rate of complete remission.

Glycyrrhetinic acid inhibits the apoptosis of gastric mucosa cells in gastric ulcer rats / 中成药

AIM To explore the effects of glycyrrhetinic acid on the gastric ulcer rats infected by Helicobacter pylori (Hp) and its action mechanism.METHODS Gastric ulcer rat models were induced by acetic acid stress and then followed by Hp infection.After treatment with low and high doses of glycyrrhetinic acid,the ulcer index,gastric acid and proteinase activities in gastric ulcer rats were analyzed.The effects of glycyrrhetinic acid on the expressions of BCL2 and Caspase-3,the GSK3β activity in gastric mucosa and gastric epithelial cells,and the cell apoptosis level were then detected.RESULTS Glycyrrhetinic acid reduced the ulcer index,gastric acid and proteinase activities in rats.Besides,the expression of BCL2 was significantly up-regulated by glycyrrhetinic acid in gastric mucosa and gastric epithelial cells,whereas the expression of Caspase-3,level of cell apoptosis,and GSK3β activity were significantly reduced.After the treatment with GSK3 β activator LY294002,the level of BCL2 was down-regulated,Caspase-3 expression was increased,and the level of cell apoptosis was enhanced.CONCLUSION Glycyrrhetinic acid promotes the healing of gastric ulcer infected by Hp via regulating GSK3β activity and inhibiting apoptosis of gastric epithelial cells.

Optimal scan time of MRI with alpha-methyl-L-tryptophan superparamagnetic iron oxide nanoparticles for temporal lobe epilepsy / 中华核医学与分子影像杂志

Objective To investigate the optimal scan time of MRI using the imaging probe alphamethyl-L-tryptophan(α-MTrp)-superparamagnetic iron oxide nanoparticles (SPIONs) for localizing temporal lobe epilepsy (TLE) foci.Methods α-MTrp-SPIONs were injected into rat models of TLE through the tail vein during the acute and chronic stages (72 h and 8 weeks after status epilepticus,respectively).MRI was performed before and 1,2,4,8,24 h after the injection in all animals,and the T2 values of the epileptogenic regions were measured.One-way repeated measures analysis of variance was used for data analysis.Results Compared with the T2 values before the injection of α-MTrp-SPIONs,the T2 signal of epileptogenic regions after the injection had a negative increased change.The T2 values before and 1,2,4,8,24 h after the injection in acute stage were 112.08±5.85,107.83±6.59,105.08±6.79,95.58±5.14,100.92± 5.81,105.17±6.31 respectively,and those in chronic stage were 112.08±7.53,107.75±7.10,102.75± 5.50,96.17±5.01,97.75±4.37,102.92±4.74.The T2 values after the injection were significantly different from those before the injection (both P＜0.01).The T2 value at 4 h after the injection decreased mostly.Conclusions α-MTrp-SPIONs can precisely localize epileptogenic regions of TLE on MRI.The optimal scan time is 4 h after the injection.

Effect of berberine on endoplasmic reticulum stress PERK apoptosis pathway in HK-2 cells by high fructose / 解放军医学杂志

Objective To investigate the effect of berberine on endoplasmic reticulum stress PERK apoptosis pathway in HK-2 cells by high fructose.Methods HK-2 cells were grown in DMEM/F12, containing 10% fetal bovine serum (FBS) and divided randomly into four groups: normal control group (Group C); Fructose group (Group F): it contains 25mmol/L fructose culture; Berberine group (Group B): 25mmol/L fructose + 10μmol/L berberine treatment group; TUDCA group (Group T):25mmol/L fructose +2μmol/L TUDCA culture group; Cells were collected after culturing 24h. The expression of glucose-regulated protein 78 (GRP78), CHOP protein and the phosphorylation levels of PERK, eIF2α were tested by Western blotting. The cell cycles were detected by flow cytometry and the apoptosis of cells were detected by TUNEL staining.ResultsWestern blotting showed that the expression of GRP78 and CHOP protein in group F was significantly higher than that in group C, and the levels of p-PERK and p-eIF2α in group F were significantly higher than those in group F. Compared with group F, GRP78, CHOP, p-PERK and p-eIF2α in group B and T were significantly lower (P0.05).Conclusion Persistent high fructose can activate the intracellular PERK pathway in HK-2 cells, causing endoplasmic reticulum stress. Berberine can inhibit the fructose-induced PERK and eIF2α phosphorylation, down-regulated the expression of GRP78, CHOP protein, thus by regulating PERK Pathways to alleviate cell cycle arrest and reduce cell apoptosis.

High-level expression and characterization of Selenomonas ruminantium β-xylosidase in Pichia pastoris / 生物工程学报

β-xylosidase (EC 3.2.1.37) is an important part of the xylanolytic enzymes system. In the present research, β-xylosidase gene Sxa derived from Selenomonas ruminantium was expressed in Pichia pastoris GS115. According to the codon bias and rare codons of P. pastoris, mRNA secondary structure and GC content, Sxa gene was optimized. The optimized full-length gene mSxa was obtained by gene synthesis technique and the recombinant yeast expression vector pPIC9K-mSxa was constructed. After being digested by restriction enzyme BglⅡ, the mSxa gene was transformed into P. pastoris GS115. Then, phenotype and geneticin G418 resistance screening, and PCR were adopted to identify the positive transformants. Finally, the recombinant P. pastoris GS115-pPIC9K-mSxa was obtained. Based on enzymatic activity assay, a high-level expression clone was picked up and then the enzymatic characteristics of the recombinant β-xylosidase were studied. The results showed that the molecular weight of the mSxa expressed in P. pastoris G115 was about 66 kDa. The maximum activity was achieved 287.61 IU/mL at fermenter level. Enzymatic characterization showed the β-xylosidase was stable between 40 ℃ and 60 ℃, and pH between 5.0 and 7.0. The optimal reaction temperature and pH were 55 ℃ and 6.0, and preferentially degrading the β-xylose glycosidic bond. The enzymatic activity was activated by Mn²⁺ and Ca²⁺, and inhibited by Fe³⁺, Cu²⁺, Co²⁺, Mg²⁺, EDTA and SDS. The study indicates that the modified β-xylosidase gene mSxa from Selenomonas ruminantium can express successfully with high activity in P. pastoris. The study lays a foundation for further industrial application of the β-xylosidase.

Artemisinin attenuates intestinal epithelial barrier damage induced by LPS / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the effect of artemisinin on lipopolysaccharide ( LPS)-induced intestinal epi-thelial barrier damage in IEC-6 cells and its molecular mechanism.METHODS:Cultured IEC-6 cells were divided to 5 groups:control group, LPS (100 mg/L) group and LPS +Artemisinin (30, 50 and 100μmol/L) groups.The cytotoxici-ty was detected by MTT assay.The releases of TNF-α, IL-1βand IL-6 in the IEC-6 cells were measured by ELISA.The transepithelial electrical resistance ( TER) was detected by electrical resistance tester, and the horseradish peroxidase (HRP) flux permeability were analyzed by a microplate reader.The expression of tight junction proteins, ZO-1, claudin-1 and occludin, and the expression of TLR4/MyD88/NF-κB at mRNA and protein levels were determined by RT-qPCR and Western blot.RESULTS:Artemisinin alone (up to 100 μmol/L) or in combination with LPS (100 mg/L) was not toxic to IEC-6 cells.Compared with control group, the releases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells significantly increased after treatment with LPS.The expression of TLR4/MyD88/NF-κB was activated by LPS.LPS down-regulated the protein expression of ZO-1, claudin-1 and occludin.However, artemisinin treatment decreased the re-leases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells.The expression of TLR4/MyD88/NF-κB at mR-NA and protein levels was gradually reduced after treatment with artemisinin.In addition, artemisinin upregulated the pro-tein expression of ZO-1, claudin-1 and occludin significantly (P<0.01) in a dose-dependent manner.CONCLUSION:Artemisinin attenuates LPS-induced intestinal epithelial barrier damage by inhibiting TLR4/MyD88/NF-κB activation in the IEC-6 cells.

FGFR1 selective inhibitor PD173074 can reduce proliferation and induce apoptosis of nasopharyngeal carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the influence of PD173074 on proliferation and apoptosis of nasopharyngeal carcinoma.@*METHOD@#With immunoblotting and RT-PCR, FGFR1 expression was detected in CNE, PONE1 and C666-1 cell lines. With MTT assay,the time-effect and dose-effect correlation between PD173074 and inhibition of CNE proliferation was evaluated. After PD173074 stimulation, the phosphorylation level of FGFR1 and AKT was detected with immunoblotting assay. Furthermore, influence of PD173074 on the activation of Caspase3 and Caspase9 was detected to study the underlying mechanism of why PD173074 could inhibit CNE proliferation.@*RESULT@#FGFR1 has the highest expression in CNE cell line. Under incubation of 10 nmol/L PD173074 stimulation for 36 hours to 72 hours, the phosphorylation of FGFR1 and AKT was impaired significantly, which further reduced the proliferation of CNE. Moreover, PD173074 can activate the intrinsic apoptotic pathway by stimulating Caspase9,which activated Caspase3 and induced the apoptosis.@*CONCLUSION@#PD173074 could inhibit proliferation of nasopharyngeal carcinoma cell through reducing the phosphorylation of FGFR1 and AKT. Additionally, PD173074 can induce CNE apoptosis by activating intrinsic apoptotic pathway via cleaving Caspase9 and Caspase3.

Two Methods for Disinfecting Slippers Contaminated in Operating Room:A Comparison / 中华医院感染学杂志

OBJECTIVE To explore the disinfection methods for slippers that used in operation room. METHODS A total of 120 pairs of slippers were monitored on their disinfection half of them were with automatic washer machine as a test group; while control group was dealed mannally with compound chlorine disinfection solution. The disinfection effects of two groups were compared. RESULTS The disinfection effects of two methods were identical, but had a difference of work efficiency. CONCLUSIONS The slippers used in operating room are contaminated by bacteria seriously. The manual disinfection method is low inefficient. The test group by machine method is with good efficiency and safety, so, it was recommendable for clinical use.