Willingness to pay integrated medical and elderly care services among the elderly in Hohhot City / 预防医学

Objective@#To investigate the willingness to pay integrated medical and elderly care services and identify the influencing factors among the elderly in Hohhot City, Inner Mongolia, so as to provide insights into promoting integrated medical and elderly care services.@*Methods@#The elderly at ages of 60 years and older were sampled using a convenient sampling method from public venues in four districts of Huimin, Saihan, Yuquan and Xincheng in Hohhot City from June to December 2021. Basic characteristics, health and disease burdens, social support and willingness to pay integrated medical and elderly care services were collected, and factors affecting the willingness to pay integrated medical and elderly care services were identified among the elderly using a multiple linear regression model. @*Results@#A total of 1 008 valid questionnaires were recovered, with an effective recovery rate of 96.74%. The respondents included 519 men (51.49%) and 489 women (48.51%), and had a mean age of (69.47±6.42) years. The monthly average fees of willingness to pay integrated medical and elderly care services were (2 076.49±36.79) Yuan, and there were 636 participants with 2 000 Yuan and less monthly average fees of willingness to pay integrated medical and elderly care services (63.10%). Multiple linear regression analysis showed that place of residence (β=180.832), satisfaction with housing (satisfied, β=-140.760), physical self-care ability (completely self-care: β=-238.244; mostly self-care: β=-254.557), burdens of disease diagnosis and treatment (able to afford: β=452.488; partly afford: β=228.626), monthly income (β=347.144), expenses of medications (β=0.019) and total score of social support (β=17.116) were factors affecting the willingness to pay integrated medical and elderly care services among the elderly. @*Conclusions@#The willingness to pay integrated medical and elderly care services among the elderly in Hohhot City is associated with place of residence, monthly income, satisfaction with housing, physical self-care ability, burden of disease diagnosis and treatment, expenses of medications and social support.

.Expression and purification of Mycobacterium tuberculosis MPT83 protein and its application in tuberculosis immunological diagnosis / 中国热带医学

@#Abstract: Objective To express and purify MPT83 protein of Mycobacterium tuberculosis and evaluate its application value in immunological diagnosis of tuberculosis (TB) using clinical samples. Methods Using Mycobacterium tuberculosis (Mtb) H37Rv genome as the template, Mtb mpt83 gene was amplified by PCR and connected to PET-21a (+) to construct prokaryotic expression vector, and then transferred into E.coli DH5α. The positive colonies were picked out and retained. The recombinant plasmid pET-mpt83 of the strain with positive colony PCR was extracted, identified by double digestion, and the samples of the positive colonies were sent for sequencing. The correctly sequenced plasmids then were transferred into BL21 competent cells for induction, expression and purification with nickel column affinity chromatography. The purified products were identified by 12 alkyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Mouse polyclonal antiserum was prepared by immunizing mice with purified protein. 8 patients clinically diagnosed as tuberculosis pleural effusion (TB group) and 8 adenocarcinomas patients (CA group) were enrolled and their pleural effusion and plasma were collected. 8 healthy people (HC group) were enrolled as the control group and their plasma were collected. An indirect ELISA was used to detect the level of specific antibodies recognizing MPT83 protein in the samples. Results Mtb MPT83 protein was successfully expressed and purified. The serum titer of MPT83 mouse polyclonal antibody was as high as 1∶1 280 000. The plasma levels of MPT83 antigen specific antibodies in TB group were significantly higher than those in HC group (P<0.05), while the plasma levels of MPT83 antigen specific antibodies in CA group were not significantly different from those in HC group (P>0.05). Compared with the HC group, there was no significant difference in pleural fluid in both the TB and CA groups (P>0.05). The ROC curve was used to analyze the OD values of plasma in TB group and HC group, and the area under the curve was greater than 0.7, showing high diagnostic efficacy. Conclusion　MPT83 protein has high antigen specificity and immunogenicity, which has great application value in the immunological diagnosis of tuberculosis.

Anatomy of the original short head of biceps-muscular structure / 解剖学报

Objective To analyze and describe the morphological characteristics of the proximal origin of the biceps brachii, and measure the anatomical data of its tendons, so as to provide references for related clinical applications of the biceps brachii. Methods Six adult shoulder specimens (4 on the left and 2 on the right) were dissected to observe the origin, morphology, and course of the biceps brachii tendon and its adjacent relationship with the coracobrachialis muscle, musculocutaneous nerve and other structures. And then the length, width, and thickness of the biceps tendon were measured. Results The origin of the short head of biceps consisted of muscle fibers attaching directly to the tip of the coracoid process, with a thin, tendinous aponeurosis covering its anterior surface, rather than a true tendon as previously described. The mean of length,widths and thickness of the short head of biceps were (107.7±9.6) mm,(20.0±1.5)mm and (5.8±2.0)mm. Conclusion The short head of the biceps brachii is directly from the coracoid process with muscle fibers.

Progress for targeting adenosine A2A receptors in cancer immunotherapy / 药学学报

Immunotherapy has completely changed the paradigm of clinical tumor treatment, but immune checkpoint inhibitors still have low objective response rates and are prone to drug resistance for most solid tumors. The immune suppression tumor microenvironment and complicated tumor immune escape mechanisms are key factors that affect the clinical outcome and response rates. Therefore, it is critical to reverse the obstacle of the tumor microenvironment to improve immunotherapy efficacy. The immune suppression caused by the increased level of adenosine in the tumor microenvironment raises the attention of people. Targeting adenosine receptors, especially A2AR, will be an effective strategy to improve immunotherapy efficacy. Targeting the adenosine-A2A pathway can increase immune infiltration, enhance immune cell function, and partially reverse immunotherapy-insensitive "cold tumors" to "hot tumors" to enhance treatment response rates and improve the efficacy of current immunotherapy. At present, many adenosine receptor inhibitors have shown good results in clinical trials, especially in combination with immune checkpoint inhibitors, chemotherapy, and adoptive cell transfer therapeutic drugs, which are expected to be used for tumor immunotherapy to bring new breakthroughs. This article reviews the accumulation mode of adenosine in the tumor microenvironment, the role of A2AR and their regulatory mechanism in immune response, the progress of A2AR inhibitors in clinical trials, potential risks to target A2AR, and the prospects for therapeutic targeting A2AR.

Genetic Mutation Characteristics of Glucose-6-Phosphate Dehydrogenase Deficiency Patients in Wuhan / 中国实验血液学杂志

OBJECTIVE@#To explore the genotype mutation characteristics of patients with glucose-6-phosphate dehydrogenase(G6PD) deficiency in Wuhan.@*METHODS@#A total of 1 321 neonates with positive screening and outpatients were received G6PD mutation detection, 12 kinds of common G6PD mutation in Chinese people was detected by using multicolor melting curve analysis (MMCA) method, for those with negative results, the enzyme activity and clinical information were analyzed, sequencing was recommended after informed consent when it is necessary.@*RESULTS@#Among 1321 patients, a total of 768 mutations were detected out, with a detection rate of 58.1%. A total of 18 types of G6PD genotypes were identified, including c.1388G>A, c.1376G>T, c.95G>A, c.1024C>T, c.871G>A, c.392G>T, c.487G>A, c.1360C>T, c.1004C>A, c.517T>C, c.592C>T, c.94C>G, c.152C>T, c.320A>G, c.1028A>G, c.1316G>A, c.1327G>C and c.1376G>C, including 683 male hemizygotes, 3 female homozygotes, 80 female heterozygotes and 2 female compound heterozygous.@*CONCLUSION@#A total of 18 types of G6PD mutations are identified in the reaserch, and c.94C>G, c.1028A>G and c.1327G>C are first reported in Chinese population. The most common G6PD mutation types in Wuhan are c.1388G>A, c.1376G>T, c.95G>A.

Relationship between chronic constipation and brain-gut microflora axis / 中华胃肠外科杂志

The role of the brain-gut microbiota axis in functional gastrointestinal diseases has been gradually recognized. According to the ROME IV diagnostic criteria, functional gastrointestinal diseases are classified as diseases caused by abnormal brain-gut interaction. This concept is of great significance to the change of diagnosis and treatment paradigm of functional gastrointestinal diseases. Chronic constipation is the most common functional gastrointestinal disease. The pathogenesis of chronic constipation is closely related to the imbalance of intestinal flora, the abnormality of enteric nervous system and neurotransmitter in brain. Therefore, in the diagnosis and treatment of chronic constipation, enough attention should be paid to the concept of integration of brain-gut microflora axis, but the clinical application of brain-gut microflora axis is still limited. This may be one of the factors for high incidence but poor treatment efficacy of chronic constipation. Based on the global research progress and our clinical experience, this article expounds the clinical significance of the brain-gut microbiota axis in chronic constipation.

Flourishing Epitranscriptomics Research / 中国生物化学与分子生物学报

So far, researchers have found more than one hundred different kinds of chemical modifications on RNA. Most of these modifications are distributed on high abundant non-coding RNAs and are important for maintaining the functions of these non-coding RNAs. In recent years, thanks to the application of high-resolution mass spectrometry and the development of whole-transcriptome sequencing technologies, more and more modifications on mRNA have been discovered, and accurately mapped and localized, including N

Competency Model of Physical Therapists in China: An Exploratory Research / 中国康复理论与实践

Objective:To construct the competency model of physical therapists in China. Methods:A set of physical therapists' competency elements was established based on literature review and expert interviews. A questionnaire was designed based on the set, and 641 rehabilitation practitioners were investigated (521 valid questionnaires returned) with the questionnaire. Exploration factor analysis was used to construct the competency model of physical therapists, and confirmatory factor analysis was used to confirmed. Results:The set of physical therapists' competency elements included 74 elements in four dimensions. There were 44 competency elements in the competency model of physical therapists, including five dimensions that were Professionalism and Moral Conduct, Knowledge and Skills of Evaluation and Intervention, Abilities of Educational-Research and Advocate, Abilities of Cooperation and Management, and Basic Knowledge of Discipline, covering 74.41% of the total variance. The Cronbach's α coefficient of the whole questionnaire and each dimension was more than 0.8. Model fitness indexes of confirmatory factor analysis were as follow: χ2/df = 2.340, Root Mean Square Error of Approximation = 0.060, Goodness of Fit Index = 0.746, Normed Fit Index = 0.811, Incremental Fit Index = 0.850, Comparative Fit Index = 0.850. Conclusion:The competency model can provide a basis for physical therapists to become independent occupation, as well as for the education, selection, evaluation of physical therapists in China.

Seed oil of Brucea javanica induces apoptosis through the PI3K/Akt signaling pathway in acute lymphocytic leukemia Jurkat cells / 中国天然药物

Brucea javanica oil emulsion (BJOE) has been used to treat tumor in China for more than 40 years. However, its components and effectiveness in the treatment of acute lymphocytic leukemia (ALL) and its mechanism of anti-cancer activity remain unknown. In the current study, high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used to analyze the components of BJOE. Then, the anti-leukemia effects of BJOE were examined both in vitro and in vivo using ALL Jurkat cells and the p388 mouse leukemia transplant model, respectively. The primary ALL leukemia cells were also used to confirm the anti-leukemia effects of BJOE. The apoptotic-related results indicated that BJOE induced apoptosis in Jurkat cells and were suggestive of intrinsic apoptotic induction. Moreover, BJOE inhibited Akt (protein kinase B) activation and upregulated its downstream targets p53 and FoxO1 (forkhead box gene, group O-1) to initiate apoptosis. The activation of GSK3β was also involved. Our findings demonstrate that BJOE has anti-leukemia effects on ALL cells and can induce apoptosis in Jurkat cells through the phosphoinositide3-kinase (PI3K) /Akt signaling pathway.

Diagnosis and treatment of chronic intestinal pseudo-obstruction / 中国实用外科杂志

Chronic intestinal pseudo-obstruction(CIPO)is a rare but serious intestinal dyskinesia characterized by impaired bowel motor function in the absence of mechanical intestinal obstruction. CIPO may be caused by primary and secondary factors that damage the enteric nervous system(neuropathy),smooth muscle(myopathy),and/or Cajal interstitial cells(interstitial disease). CIPO is extremely easy to missed diagnosis and misdiagnosis.The treatment of CIPO is very difficult.It is based on nutritional support,medicine and surgical treatment.At present,the occurrence,development and treatment of intestinal micro-ecology in CIPO has gradually attracted attention and may become a new treatment method for CIPO.

Effect analysis of 71 cases of refractory diarrhea with severe malnutrition treated by fecal microbiota transplantation(FMT) combined with enteral nutrition / 中国实用外科杂志

OBJECTIVE: To investigate the therapeutic effect of fecal microbiota transplantation(FMT) combined with enteral nutrition on patients with refractory diarrhea and severe malnutrition.METHODS: Seventy-one patients with refractory diarrhea complicated with severe malnutrition in Department of Colorectal Disease,the Tenth People ' s Hospital of Tongji University and Department of General Surgery,General Hospital of Eastern Theater Command from June 2014 to October 2018 were prospectively collected for basic information,symptoms and signs,number of bowel movements,Bristol stool scores,and nutritional indicators,which were followed up after bacterial transplantation combined with enteral nutrition.RESULTS: After following up for 6 month,FMT combined with enteral nutrition in the treatment of severe diarrhea with severe malnutrition,the effective rate of treatment of diarrhea was 95.8%,of which the cure rate was 74.7%,and the remission rate was 21.1%.3 cases(4.2%) were ineffective,6 cases(8.5%) recurred.The frequency of bowel movements decreased from(8.3±2.2) before treatment to(5.3±1.3) after 1 month,(3.2±0.8)after 3 months,and(1.9±0.6) after 6 months.The Bristol score was reduced from(6.6±0.3) points before treatment to(4.8±0.7) points 1 month after treatment,(4.6±0.8)points after 3 months,and(4.3±0.8) points after 6 months.The body mass index(BMI) of the patients before treatment was 15.3±1.3,the BMI increased to 17.4±2.4 after 1 month of treatment,increased to 19.0±3.2 after 3 months,and 19.3±3.8 after 6 months.The total protein was increased from(48.6±4.7)g/L before treatment to(55.4±5.7)g/L after1 month of treatment,increased to(60.3±5.4) g/L after 3 months,and(68.4±6.9)g after 6 months.Albumin increased from(22.4±1.7)g/L before treatment to(31.7±2.4)g/L after 1 month of treatment,increased to(35.8±2.7)g/L after 3 months,and(37.5±3.7)g/L after 6 months.Fibrinogen increased from(1.6±0.4)g/L before treatment to(2.0±0.7)g/L after 1 month of treatment,increased to(2.4±0.8) g/L after 3 months,and(2.4±0.7) g/L after 6 months.Prealbumin increased from(82.0±18.6)mg/L before treatment to(178.3±25.3)mg/L after 1 month of treatment,increased to(235.7±28.2) mg/L after 3 months,and(259.6±33.5) mg/L after 6 months.The were significant differences before and after treatment(All P<0.05) CONCLUSION: After 6 months of follow-up,FMT combined with enteral nutrition can effectively reduce the number of diarrhea,improve stool characteristics,and improve nutritional indicators such as BMI,total protein,albumin,fibrinogen and prealbumin.

Effects of hydrogen sulfide on renal fibrosis in diabetic rats and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of hydrogen sulfide (HS) on renal fibrosis in diabetic rats and explore its mechanism.@*METHODS@#Male Sprague-Dawley rats were randomly divided into normal control (NC) group, a diabetic control (DC) group, diabetes mellitus (DM)+sodium hydrosulfide (NaHS) group and DM+DL-propargylglycine (PAG) group, with 8 rats in each group.Type 1 diabetes was induced in the respective groups by a single intraperitoneal (i.p.) injection of streptozotocin.From the fifth week, rats in the DM+NaHS and DM+PAG groups were injected (i.p.) with 56 μmol/kg NaHS and 40 mg/kg PAG once a day, respectively.After treatment for 4 weeks, the levels of fasting blood glucose (FBG), blood urea nitrogen (BUN) and serum creatinine (SCr) were detected.The deposition of renal collagen fibers was observed by Masson staining, and collagen volume fraction (CVF) was calculated.The ultrastructural change of renal tissue was observed by transmission electron microscopy.The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and hydroxyproline (Hyp) in renal tissues were detected using the kits.The expression levels of TGF-β1, Smad3, phosphorylated (p)-Smad3 and collagen-IV (col-IV) in renal tissues were detected using Western blot.@*RESULTS@#Compared with the NC group, the levels of FBG, BUN, SCr, CVF, IL-1β, IL-6, TNF-α and Hyp were increased; the deposition of renal collagen fibers and the ultrastructural damage were aggravated; the levels of TGF-β1, Smad3, p-Smad3, p-Smad3/Smad3 and col-IV were increased in the DC group.Compared with the DC group, excluding FBG, the aforementioned indices were improved in the DM+NaHS group; the aforementioned indices were further aggravated in the DM+PAG group.@*CONCLUSIONS@#HS attenuated renal fibrosis in diabetic rats, and the mechanism might be associated with the reduction of the release of proinflammatory cytokines, downregulation of the TGF-β1/Smad3 pathway, and inhibition of excessive accumulation of col-IV.

Lumbar Scoliosis Induction in Juvenile Dogs by Three-dimensional Modulation of Spinal Growth Using Nickel-Titanium Coil Springs / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Current treatments for scoliosis have some defects and complications. To study spinal deformities and test novel scoliosis treatments, many animal models of scoliosis have been developed. These models applied a single load to the spine and could not precisely modulate the spinal growth in different dimensions. In this study, we applied posterior tethering in various directions with the application of nickel-titanium (NT) coil springs in dog's spine to modulate spinal growth in the coronal, sagittal, and transverse planes and create a scoliosis model possess curves that mimic adolescent idiopathic scoliosis (AIS) three dimensionally.</p><p><b>METHODS</b>Scoliosis was surgically induced in eight 8-week-old female dogs (weight: 1.95-2.30 kg) using bone screws and NT coil springs. The deformity was induced through the placement of posterior NT coil springs that tethered the spine by bone screw fixation. All dogs were monitored with serial radiographs to document changes in deformities.</p><p><b>RESULTS</b>All experimental animals developed scoliotic curves convex to the left in the lumbar segment. The mean coronal Cobb angle was 18.0° immediately postoperatively and 54.5° at 22 weeks. The mean lordosis increased from 6.2° postoperatively to 35.0° at final follow-up. Apical axial rotation increased from 4.5° postoperatively to 31.2° at 22 weeks.</p><p><b>CONCLUSIONS</b>With the application of NT springs in dogs that allowed posterior tethering in various directions, lumbar spinal deformity was achieved in three planes: coronal, sagittal, and transverse planes. Notably, the lumbar spine in surgically treated dogs developed lordoscoliosis with obvious rotation and the curves mimic AIS three dimensionally well. This method allows lumbar scoliosis to develop without deep dissection of muscle and maintains the essential anatomical elements along the spinal curve. Moreover, the spinal growth modulation technique could yield information that would provide a basis for developing novel early-stage treatments for children with scoliosis.</p>

Recent Advances in Technique and Clinical Outcomes of Minimally Invasive Spine Surgery in Adult Scoliosis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>Conventional open spinal surgery of adult scoliosis can be performed from anterior, posterior, or combined approach. Minimally invasive spine surgery (MISS) was developed for the purpose of reducing the undesirable effects and complications. This review aimed to make a brief summary of recent studies of the approach and clinical outcomes of MISS in adult scoliosis.</p><p><b>DATA SOURCES</b>We conducted a systematic search from PubMed, Medline, EMBASE, and other literature databases to collect reports of surgical methods and clinical outcomes of MISS in treatment of adult scoliosis. Those reports were published up to March 2017 with the following key terms: "minimally invasive," "spine," "surgery," and "scoliosis."</p><p><b>STUDY SELECTION</b>The inclusion criteria of the articles were as followings: diagnosed with adult degenerative scoliosis (DS) or adult idiopathic scoliosis; underwent MISS or open surgery; with follow-up data. The articles involving patients with congenital scoliosis or unknown type were excluded and those without any follow-up data were also excluded from the study. The initial search yielded 233 articles. After title and abstract extraction, 29 English articles were selected for full-text review. Of those, 20 studies with 831 patients diagnosed with adult DS or adult idiopathic scoliosis were reviewed. Seventeen were retrospective studies, and three were prospective studies.</p><p><b>RESULTS</b>The surgical technique reported in these articles was direct or extreme lateral interbody fusion, axial lumbar interbody fusion, and transforaminal lumbar interbody fusion. Among the clinical outcomes of these studies, the operated levels was 3-7, operative time was 2.3-8.5 h. Both the Cobb angle of coronal major curve and evaluation of Oswestry Disability Index and Visual Analog Scale decreased after surgery. There were 323 complications reported in the 831 (38.9%) patients, including 150 (18.1%) motor or sensory deficits, and 111 (13.4%) implant-related complications.</p><p><b>CONCLUSIONS</b>MISS can provide good radiological and self-evaluation improvement in treatment of adult scoliosis. More prospective studies will be needed before it is widely used.</p>

Expression characteristics of the USP24 gene in the mouse testis during spermatogenesis / 中华男科学杂志

Objective@#To investigate the expression characteristics of the USP24 gene in the mouse testis and its role in spermatogenesis.@*METHODS@#We examined the expression characteristics of USP24 in the testis tissues of wild-type mice at different postnatal weeks (PNW) and androgen receptor (AR)-knockout (ARKO) adult mice using real-time quantitative PCR and immunofluorescence, and detected the transcriptional activity of the USP24 promoter by dual-luciferase reporter gene assay.@*RESULTS@#The expression of the USP24 gene was low in the testis tissue of the wild-type mice at PNW 1, increased dramatically at PNW 3 and stayed at a similar level till PNW 8. The USP24 protein was located mainly in the cytoplasm of Sertoli and spermatogenic cells. Compared with the wild-type, the adult ARKO mice showed a decreased expression of USP24 localized in the posterior head and mid-piece of the mature sperm in the testis. Dual-luciferase reporter gene assay showed that the transcriptional activity of the USP24 promoter was increased after testosterone stimulation.@*CONCLUSIONS@#The increased expression of the USP24 gene was associated with the initiation of sexual development, and the USP24 protein was expressed in the mature sperm of the mice. USP24 is an AR-target gene, which may be involved in the regulation of spermatogenesis in mice.

Implementation of a Pediatric Emergency Triage System in Xiamen, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pediatric emergency rooms (PERs) in Chinese hospitals are perpetually full of sick and injured children because of the lack of sufficiently developed community hospitals and low access to family physicians. The aim of this study was to evaluate the clinical value of a new five-level Chinese pediatric emergency triage system (CPETS), modeled after the Canadian Triage System and Acuity Scale.</p><p><b>METHODS</b>In this study, we compared CPETS outcomes in our PER relative to those of the prior two-level system. Patients who visited our PER before (January 2013-June 2013) and after (January 2014-June 2014) the CPETS was implemented served as the control and experimental group, respectively. Patient flow, triage rates, triage accuracy, wait times (overall and for severe patients), and patient/family satisfaction were compared between the two groups.</p><p><b>RESULTS</b>Relative to the performance of the former system experienced by the control group, the CPETS experienced by the experimental group was associated with a reduced patient flow through the PER (Cox-Stuart test, t = 0, P < 0.05), a higher triage rate (93.40% vs. 90.75%; χ2 = 801.546, P < 0.001), better triage accuracy (96.32% vs. 85.09%; χ2 = 710.904, P < 0.001), shorter overall wait times (37.30 ± 13.80 min vs. 41.60 ± 15.40 min; t = 11.27, P < 0.001), markedly shorter wait times for severe patients (2.07 [0.65, 4.11] min vs. 3.23 [1.90,4.36] min; z = -2.057, P = 0.040), and higher family satisfaction rates (94.23% vs. 92.21%; χ2 = 321.528, P < 0.001).</p><p><b>CONCLUSIONS</b>Implementing the CPETS improved nurses' abilities to triage severe patients and, thus, to deliver the urgent treatments more quickly. The system shunted nonurgent patients to outpatient care effectively, resulting in improved efficiency of PER health-care delivery.</p>

Correlation between uncoupling protein 2 expression and myocardial mitochondrial injury in rats with sepsis induced by lipopolysaccharide / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the correlation between uncoupling protein 2 (UCP2) expression and myocardial mitochondria injury in rats with sepsis induced by lipopolysaccharide (LPS).</p><p><b>METHODS</b>The rat model of sepsis was established through an intraperitoneal injection of LPS. Forty male Sprague-Dawley rats were randomly and equally divided into control group (an intraperitoneal injection of normal saline), sepsis 6 h group (LPS-6 h group), sepsis 12 h group (LPS-12 h group), sepsis 24 h group (LPS-24 h group), and sepsis 48 h group (LPS-48 h group). The serum and heart tissues were harvested at corresponding time points and myocardial mitochondria was extracted. The microplate reader was applied to measure creatine kinase (CK), creatine kinase-MB (CK-MB), and reactive oxygen species (ROS). Flow cytometry was applied to measure the degree of mitochondrial swelling and mitochondrial membrane potential (MMP). Western blot was used to measure the expression level of UCP2. Electron microscopy was applied to observe the morphological changes in heart tissues and myocardial mitochondria.</p><p><b>RESULTS</b>Compared with the control group, the LPS groups had significantly increased serum levels of CK, CK-MB, and myocardial ROS, as well as a significantly increased degree of mitochondrial swelling (P<0.05), and these values reached their peaks at 24 hours after LPS injection. The LPS groups had a significant decrease in MMP (P<0.05), which reached the lowest level at 24 hours after LPS injection. Western blot showed that the LPS groups had a significant increase in the expression level of myocardial UCP2 compared with the control group (P<0.05), which reached its peak at 24 hours after LPS injection. The results of electron microscopy showed mitochondrial swelling, partial rupture of the mitochondrial membrane, and cavity formation in rats in the LPS groups. The most severe lesions occurred in the LPS-24 h group. In rats with LPS, the ROS level in the myocardial mitochondria and the degree of mitochondrial swelling were positively correlated with the expression level of UCP2 (r=0.796 and 0.893, respectively; P<0.05), while MMP was negatively correlated with the expression level of UCP2 (r=-0.903, P<0.05).</p><p><b>CONCLUSIONS</b>In the rat model of sepsis, the myocardium and myocardial mitochondria have obvious injuries, and the expression level of UCP2 is closely correlated with mitochondrial injury. Therefore, UCP2 might play an important role in myocardial mitochondrial injury in sepsis.</p>

Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. Methods IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice. Results Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg). Conclusions IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.

Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

Background Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. Methods IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice. Results Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg). Conclusions IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.(AU)

A preliminary study of long-term mitochondrial dysfunction in rat brain caused by lipopolysaccharide-induced sepsis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To preliminarily investigate the long-term structural and functional injuries of mitochondria in rat brain caused by sepsis.</p><p><b>METHODS</b>Wistar rats were randomly assigned into sepsis and control groups. A rat model of sepsis was prepared by an intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) of gram-negative bacteria, and the survival assay was performed. Eight rats in the sepsis group were sacrificed at 12, 24, 48, or 72 hours after LPS injection, while rats in the control group were sacrificed after an intraperitoneal injection of an equal volume of normal saline. Mitochondria were extracted from rat brain tissue. Mitochondrial membrane potential (MMP) and mitochondrial swelling level were determined by flow cytometry, and the activities of electron transport chain complexes (I-V) were measured using enzyme assay kits. Hematoxylin-eosin (HE) staining and electron microscopy were used to observe morphological changes in brain tissue and mitochondria.</p><p><b>RESULTS</b>The sepsis group had a significantly lower survival rate than the control group (P<0.01). The MMP and activities of electron transport chain complexes (I-V) in the sepsis group, which were significantly lower than those in the control group (P<0.05), were reduced to the lowest levels at 48 hours and partially recovered at 72 hours. The mitochondrial swelling level in the sepsis group, which was significantly higher than that in the control group (P<0.05), increased to the peak level at 48 hours and partially recovered at 72 hours. Hematoxylin and Eosin staining revealed substantial damages in the structure of brain tissue, and electron microscopy showed mitochondrial swelling, and vacuolization in a few mitochondria.</p><p><b>CONCLUSIONS</b>In the rat model of LPS-induced sepsis, both structural and functional injuries are found in cerebral mitochondria, and achieve the peak levels probably at around 48 hours.</p>