Structure-based development of potent and selective type-II kinase inhibitors of RIPK1

Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) functions as a key regulator in inflammation and cell death and is involved in mediating a variety of inflammatory or degenerative diseases. A number of allosteric RIPK1 inhibitors (RIPK1i) have been developed, and some of them have already advanced into clinical evaluation. Recently, selective RIPK1i that interact with both the allosteric pocket and the ATP-binding site of RIPK1 have started to emerge. Here, we report the rational development of a new series of type-II RIPK1i based on the rediscovery of a reported but mechanistically atypical RIPK3i. We also describe the structure-guided lead optimization of a potent, selective, and orally bioavailable RIPK1i, 62, which exhibits extraordinary efficacies in mouse models of acute or chronic inflammatory diseases. Collectively, 62 provides a useful tool for evaluating RIPK1 in animal disease models and a promising lead for further drug development.

Effects of metabolic and bariatric surgery on bone metabolism in patients with obesity: A meta-analysis / 国际外科学杂志

Objective:To systematically evaluate the effect of bariatric and metabolic surgery on bone metabolism in obese patients.Methods:Search terms for the present meta-analysis included "bariatric surgery, metabolic surgery, sleeve gastrectomy, gastric bypass, bone metabolic indicators, bone mineral density", both in English and corresponding Chinese. PubMed, WOS, Cochrane, CNKI, and VIP databases were searched for longitudinal studies from the establishment of the database to September 20, 2022. The data on bone mineral density and bone metabolic markers in obese patients before and after bariatric surgery were extracted. RevMan5.4 and Stata17.0 software were used for Meta-analysis.Results:A total of 8 clinical studies with 420 patients were included. The results of the meta-analysis showed that compared with the preoperative baseline, lumbar spine bone mineral density ( WMD=0.05, 95% CI: -0.00~0.1), femoral neck bone mineral density( WMD=0.10, 95% CI: 0.05-0.15), hip bone mineral density( WMD=0.14, 95% CI: 0.10-0.17), and serum vitamin D 3 ( WMD=-4.87, 95% CI: -6.34--3.40)were decreased, while parathyroid hormone ( WMD=10.04, 95% CI: 5.32-14.76) was elevated after surgery. Conclusions:Current evidence demonstrates that metabolic and bariatric surgery can lead to decreased bone mineral density and impairs in bone metabolic markers early after surgery. Roux-en-Y gastric bypass surgery cause more adverse effects on bone metabolism than sleeve gastrectomy. The results imply that all patients undergoing metabolic and bariatric surgery should be monitored for bone metabolism and routinely take vitamin D and calcium supplements.

Research progress on the role of microbiota in radiotherapy-induced injury / 中华放射肿瘤学杂志

Human microbiota is composed of bacteria, viruses, fungi and other microorganisms, which are distributed in the oral cavity, nasal cavity, intestine, vagina and skin, etc. Human microbiota plays a pivotal role in the metabolism, immunity, hormones and homeostasis of the host. It can protect the host and maintain the homeostasis, and provoke the incidence of inflammation and tumors. Microbiota has been found to modulate the efficacy and toxicity of chemotherapy and immunotherapy for certain types of tumors. Nevertheless, large-scale studies in the context of radiation therapy have not been performed. In this article, the relationship between the microbiota and the radiotherapy response and toxicity changes of cancer patients was summarized, aiming to develop the optimal treatment plan for patients, and provide evidence for the prevention and treatment of radiotherapy-induced injury.

Diagnosis and treatment of subarachnoid hemorrhage / 中华神经科杂志

Subarachnoid hemorrhage (SAH) is a common hemorrhagic cerebrovascular disease with high mortality and disability rate. Intracranial aneurysm is the major cause of SAH,severe and unbearable headache with meningeal irritation is the typical clinical manifestation of it,and the diagnosis is mainly confirmed by neuroimaging. The purpose of SAH treatment is to eradicate bleeding causes such as aneurysms and brain arteriovenous malformation (bAVM), eliminate brain edema and control intracranial pressure, prevent cerebral vasospasm and secondary cerebral ischemia, and deal with complications such as hydrocephalus actively. Two international multicenter randomized controlled trials, ISAT (International Subarachnoid Aneurysm Trial) and BRAT (Barrow Ruptured Aneurysm Trial), have provided new evidences for the treatment of aneurysmal subarachnoid hemorrhage. Multidisciplinary collaboration,consisting of neurosurgery, intervention and neurology departments, is an important guarantee for good clinical outcomes.

Screening and identification of hepatitis B surface antigen binding to lactoferrin in human milk / 中华围产医学杂志

Objective Human milk of mothers with positive hepatitis B surface antigen (HBsAg) contains hepatitis B virus (HBV). However, breastfeeding does not increase the risk of mother-to-infant transmission of HBV. Previous investigations demonstrated that breast milk has a property of binding with HBsAg. This study aimed to identify the component in human milk that can bind to HBsAg. Methods This study was performed in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, from June 2015 to February 2017. Human milk samples from two postpartum women with negative HBV markers and two control samples of cow milk and goat milk were analyzed by Far-Western blot, in which highly purified recombinant yeast HBsAg was used to bind with whey proteins. Based on the results of mass-spectrum analysis, competition inhibition test was used to confirm the functioning component. Results Far-Western blot showed remarkable protein bands at the relative molecular weight of about 80 000 in both lanes of human milk, but none in the lane of cow or goat milk. Mass-spectrum analysis of the protein band indicated there were proteins sharing 28.4%-93.4% homology in amino acid sequences with five proteins with the highest homology to lactoferrin (93.4%). Further Far-Western blot with purified recombinant lactoferrin showed that lactoferrin could bind to the recombinant HBsAg. Competition inhibition test suggested that the purified recombinant lactoferrin inhibited the binding of HBsAg to its antibody in a dose-dependent manner. Conclusions This study confirms the capability of lactoferrin in human milk to combine with HBsAg, suggesting that lactoferrin can bind to HBV. Further study on whether lactoferrin can inhibit the infectivity of HBV would be valuable to clarify the reason for not increasing the risk of mother-to-infant transmission of HBV by breastfeeding.

A modified suction device with negative pressure for cerebrospinal fluid collection in rats / 中国比较医学杂志

Objective To improve the success rate of cerebrospinal fluid (CSF) collection in rats using a modified suction device with negative pressure. Methods A flow regulator was added to the original apparatus to adjust the intensity of negative pressure and to limit the volume of CSF collected each time. A total of 36 male Sprague- Dawley (SD) rats were randomly divided into 3 groups, and the CSF was collected in 3 ways: drawing from the cerebellomedullary cistern via percutaneous puncture, using a suction device with negative pressure and using a modified device. Kruskal-Wallis test was used to evaluate the differences in result between these three methods. Results The modified suction device with negative pressure effectively reduced bleeding. The quality of CSF collected with it was 75％ of 12 rats, significantly higher than that obtained with the original device (33. 3％, P ＜ 0. 01) or drawn from the cerebellomedullary cistern via percutaneous puncture (0％, P ＜ 0. 001). The success rate of qualified sample collection was highly superior to those of previously reported CSF collection methods. Conclusions Our modified suction device for CSF collection is simple and user-friendly, with minimal damage and little bleeding to rats. It can obviously improve the success rate of CSF collection in rats.

The expression of plasma exosomal miR-422a in ischaemic stroke and its diagnostic value as biomarker / 中华检验医学杂志

Objective To explore the potential predictive value of plasma exosomal miR-422a in different time after ischaemic stroke (1, 2, 3, 4 day) .Methods Retrospective study.Forty patients diagnosed with ischaemic stroke ( IS ) and ten age and sex matched people who underwent a standard physical examination were recruited from the First Affiliated Hospital of Guangxi Medical University between May 2016 and December 2016.Plasma exosomes were extracted by use of related kit and the expression level of plasma exosomal miR-422a in both IS patients (1, 2, 3, 4 day) and healthy controls were examined via quantitative real-time polymerase chain reaction ( qRT-PCR ) .The areas under the curve ( AUC ) of the receiver operating characteristic curve were constructed to evaluate the diagnostic accuracy of the miR -422a in IS, and one-way analysis of variance followed by the Games-Howell post hoc test was used for difference analysis.Results The exosomes isolated from human plasma showed round or oval vesicles , the average particle size was 128.2 nm and with maximum peak distribution of 186.9 nm.Moreover , all of the isolated exosomes were positive for a marker , with the CD63-positive rate at 80.6％ and the CD81-positive rate at 91.7％.qRT-PCR confirmed that miR-422a was expressed in human plasma exosomes , and the expression levels of plasma exosomal miR-422a [median relative values, 1.221 (95％CI:0.640-1.802) for healthy group, 4.418 (95％CI:2.642-6.193) for group of 1 dayafter IS, 2.912 (95％CI:2.262-3.562) for 2 day, 2.744 (95％CI:2.000-3.487) for 3 day and 0.449 (95％CI:0.170-0.727) for 4 day] were significantly increased in the time after IS 1, 2, 3 day ( F=13.57, P<0.05, P<0.01, P<0.05, respectively;and the AUC values were 0.920, 0.945, 0.870, respectively ) .The expression of plasma exosomal miR-422a on the 4th day after IS were lower than those in other IS groups ( F=13.57,P<0.005, P<0.001, P<0.001, respectively), and no statistical significance was found between the expression of plasma exosomal miR-422a on the 4th day after IS than that in the control group [0.449 (95％CI:0.170-0.727)].Conclusion Plasma exosomal miR-422a showed high diagnostic value as blood-based biomarker for diagnosing IS in the early phase (1-3 d).

Nutlin-3 promotes pyroptosis in SMMC-7721 cells / 中国免疫学杂志

Objective:To evaluate the effect of Nutlin-3,a MDM2 antagonist,on the pyroptosis in SMMC-7721 cells.Methods: The expression of actived caspase-1(p20) and IL-1β was detected using Western blot analysis.Pyroptosis was investigated by a standard lactate dehydrogenase release assay(LDH).IL-1β content in the cell culture supernatant was quantified by ELISA.Results: Nutlin-3 up-regulated the expression level of actived caspase-1 and IL-1β in SMMC-7721 cells.Meanwhile,Nutlin-3 increased significantly the content of LDH and IL-1β in the cell culture supernatant(P<0.05).Conclusion: Nutlin-3 activated pro-caspase-1,promoted pyroptosis and IL-1β release in SMMC-7721 cells.

Characterization of binding capability of human breast milk to hepatitis B surface antigen / 中华围产医学杂志

Objective To investigate whether human breast milk may bind to hepatitis B surface antigen (HBsAg) and its characteristics.Methods Breast milk samples from five women with negative HBsAg and hepatitis B surface antibody (anti-HBs) at one to two months post delivery were fractioned into cream and skimmed milk by centrifugation.The human breast milk and each fraction as well as cow and goat milk samples,served as controls,were separately incubated with highly purified yeast recombinant HBsAg,followed by determination of their binding capability to HBsAg by enzyme linked immunosorbent assay (ELISA) and the inhibition rate for binding of HBsAg to anti-HBs by quantitative chemiluminescence microparticle immunoassay.After boiled for 1 min or pasteurized in 65 ℃ for 30 min,the thermal stability of the active components of milk was detected.One-way ANOVA and SNK tests were performed for statistical analysis.Results The operative concentration of HBsAg was 0.1 μg/ml.Breast milk from all five women showed significantly better binding capability to HBsAg than cow or goat milk (1.306±0.300 vs 2.157±0.150 and 2.232±0.093,F=34.303,P＜0.01).The quantitative experiments showed that the inhibition rate of human breast milk was higher than that of the control group [(74.26± 17.26)％ vs (0.00±5.50)％,F=57.806,P＜0.01].The binding ability to HBsAg of skimmed milk was comparable with that of whole milk,indicating milk protein(s) played critical roles in binding to HBsAg (0.877 ± 0.486 vs 0.513 ± 0.069 and 0.376 ± 0.146,F=44.475,P＜0.01).After boiled for 1 min or Pasteurization,the binding ability to HBsAg of whole breast milk remained,but that of skimmed milk went down (F=16.598,P＜0.01).Both whole breast milk and skimmed milk could inhibit the binding of HBsAg to anti-HBs (F=278.341 and 269.408,both P＜0.01).Conclusions The inhibition of binding to HBsAg by human breast milk indicates that human milk may interact with HBsAg.The active components mainly exist in milk proteins and are thermal stable.

Clinical significance of paraneoplastic neurological antibodies detection / 中华检验医学杂志

Paraneoplastic neurological syndromes (PNS) were defined as neurological syndromes associated with cancer.In many cases,autoantibodies against neural antigens expressed by the tumor (paraneoplastic neurological antibody or onconeural antibodies) were detected.The neuronal antibodies,which were associated with syndromes resulting from central nervous system neuronal dysfunction,were classified into two groups according to the location of the antigen:inside the neuron or in the cell membrane.Group Ⅰ antibodies targeted on intracellular antigens and were predominantly dependent on T-cell-mediated responses.Group Ⅱ antibodies recognized neuronal surface antigens and were predominantly dependent on B-cell-mediated responses.Onconeural antibodies were useful diagnostic markers of the brain disease,and in some cases,they might reveal an underlying malignancy.Moreover,the presence of antibodies against surface(group Ⅱ) antigens might predict a more favorable response to immunotherapy than that against intracellular (group Ⅰ) antigens.

Influence of amniocentesis on risk of mother-to-child transmission of hepatitis B virus / 中华围产医学杂志

Objective To investigate whether amniocentesis may increase the risk for mother-tochild transmission of hepatitis B virus (HBV).Methods Totally 40 children born to HBV-infected mothers who had amniocentesis performed in Nanjing Drum Tower Hospital, Nanjing University Medical School from January 2010 to December 2013, were followed up and screened for HBV markers.Amniotic fluid samples were collected and stored at-80 ℃ were tested for HBV markers.Among the 40 carrier mothers, three (7.5％) were hepatitis B e antigen (HBeAg)-positive.Relevant data such as antiviral history, administration of hepatitis B vaccine and hepatitis B immunoglobulin (HBIG) in infants were collected.Chi-square test or Fisher's exact test was used for statistical analysis.Results The mothers were 21-41 years old, with a mean age of (31.5±5.5) years at the time of amniocentesis and mean gestational age of (21.2± 1.6) weeks (18.4-24.9 weeks).Indications for amniocentesis were mainly abnormal maternal serum alpha-fetoprotein levels (65.0％, 26/40)and maternal age over 35 years (10.0％, 4/40).None of the mothers received antiviral therapy and 14 (35.0％)underwent transplacental amniocentesis.Among 28 cases who had a store of amniotic fluid sample and were followed-up, one (7.1％) was positive for both hepatitis B surface antigen (HBsAg) and HBV DNA, and another was positive for HBsAg only.The average age of 40 children at follow-up was (2.0± 1.0) years (seven months to four years old), among which 23 were boys and 17 were girls.All of them received hepatitis B vaccine and HBIG.Positive rate of HBsAg and HBV DNA in HBeAg(+) mothers are higher than those in HBeAg(-) mothers [4.7％(2/43) vs 3/5, x2=14.705;0/43 vs 2/5, x2=17.948;both P ＜ 0.05].Thirty-seven children born to HBsAg(+)/HBeAg(-) mothers were negative for both HBsAg and hepatitis B core antibody (anti-HBc), and the other three born to HBsAg(+)/HBeAg(+) mothers were also negative for HBsAg and anti-HBc.Additionally, the positive rate of hepatitis B surface antibody in children was 90.0％ (36/40).Conclusions For those HBsAg(+)/HBeAg(-) mothers, amniocentesis does not elevate the risk for mother-to-child transmission of HBV, even performed transplacentally.However, further studies are needed for HBeAg-positive mothers.HBsAg or HBV DNA positive in amniotic fluid should be diagnosed as intrauterine infection of HBV.

Role of phosphorylation of p53 at Ser392 in cancer therapy / 解剖学报

Phosphorylation is the most common way of p 53 post-translational modifications .However , gaps still exist in our knowledge regarding the role and mechanism of phosphorylation of p 53 at Ser392 in carcinogenesis and cancer prevention.In the present study, we summarized the effect of phos-p53-Ser392 to wild-type and mutant p53, the regulation by DNA damage agents and protein kinase , and the significance of phosphorylation of p 53-Ser392 in cancer treatment .

Clinical applications of blood biomarkers in ischemic stroke / 中华检验医学杂志

Early diagnosis and timely treatment are two critical factors in reducing the cerebral damage and improving prognosis for ischemic stroke patients.The only proved effective way is to intravenously administer recombinant tissue plasminogen activator for thrombolytic therapy.However,the narrow time window restricts its application.One rapid and effective blood test,or auxiliary examinations such as computer tomography which may aid early diagnosis of ischemic stroke would contribute to treatment and intervention promptly,then take a pivotal role for the disease prognosis.Herein,the reported proteins and RNA markers are summarized and analysed in clinical application of acute ischemic stroke likewise.

Role of miR-184 in oxygen-glucose-deprivation induced SK-N-SH cell ischemic injury and its regulation on AKT2 level / 实用医学杂志

Objective To explore the role of miR-184 in Oxygen-Glucose-Deprivation (OGD) induced SK-N-SH cell ischemic injury and its regulation on AKT2 level. Method We used a combination of oxygen and glucose deprivation to imitate ischemic conditions in vivo. MiR-184 mimic and inhibitor were transfected into SK-N-SH cell to alter miR-184 levels. The expression of miR-184 and AKT2 were determined by using Real-time PCR. The extent of SK-N-SH cell survival rate was assessed by thiazolyl blue tetrazolium bromide (MTT) assay. Result Here, we observed that miR-184 was significantly inhibited in SK-N-SH cell after OGD (P<0.05). The changes of miR-184 level altered the expression of AKT2 mRNA. In addition, alteration of miR-184expressionsignificantly affected cell survival rate after OGD. Conclusion miR-184 plays an important role in ischemic injury through negatively regulating AKT2 level, which may provide a potential therapeutic target for ischemic stroke in miRNA levels.

DNA barcoding of rodent in Ningxia Hui Autonomous Region,China / 中国人兽共患病学报

ABSTRACT:To improve the ability of correct classification and identification of rodents in different kinds of foci of zoonosis in Ningxia Hui Autonomous Region in China ,rodents samples from different habitats of Ningxia were collected .Sequences of COI gene were amplified and sequenced from 154 samples .Based on these sequences ,the pairwise genetic distance were calcu‐lated ,and a Neighbor Joining tree were constructed .According to the NJ tree ,20 clusters with high bootstrap support were found from 19 morphological species .The striped dwarf hamsters were divided into two clusters ,which suggested that there were two cryptic species with stripe on the back .The pika from Helan Mountain showed close relationship with Ochotona pal‐lasi ,and the genetic distance was as low as 3 .6% .Results show that DNA barcodes could be used to accurately identify speci‐mens of rodents and correct morphological identification errors .It could discover appearance indistinguishable implied species and could better study the classification and evolution of rodents .

Determination of Atractylodin in Essential Oil of Jingfukang Preparations and Stability in Different Solutions / 中国中医药信息杂志

Objective To establish a method for determination of atractylodin in essential oil of Jingfukang preparations by GC and analyze its stability in different solutions. Methods Using HP-5 quartz capillary column (30 m×0.32 mm), programmed temperature was conducted with the FID as detector. The content of atractylodin was determined both at room temperature and 4 ℃ in 20 days. Results The linear range of atractylodin was 4.172-41.72 μg/mL (r=0.999 7). The stability of atractylodin in essential oil of atractylodes was better than in anhydrous alcohol solution or in essential oil of Jingfukang preparation. Conclusion This method is suitable for the quality control of atractylodin in essential oil of Jingfukang preparations. Low temperature is helpful to the stability of atractylodin. The storage period of essential oil of Jingfukang preparations should not be too long.