Inhibition of microRNA-543 alleviates sepsis-induced acute kidney injury via targeting Bcl-2.

OBJECTIVE: Sepsis has a high morbidity and mortality and is prone to cause acute kidney injury (AKI). Here, we aimed to demonstrate the function and molecular mechanism of microRNA-543 (miR-543) in septic AKI. MATERIALS AND METHODS: MiR-543 inhibitor or NC was transfected into LPS-treated HK-2 cells to observe lipopolysaccharide (LPS)-induced inflammation and apoptosis. The detection of inflammation and apoptosis of HK-2 cells relies on Western blot, quantitative Reverse-Transcription Polymerase Chain Reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), Cell Counting Kit-8 (CCK-8) assay, flow cytometry, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining. RESULTS: MiR-543 expression was increased in LPS-treated HK-2 cells. By transfecting miR-543 inhibitor into HK-2 cells, miR-543 expression was dramatically reduced. The downregulation of miR-543 remarkably inhibited the inflammation and apoptosis, which was manifested by the reduction of inflammatory cytokines (TNF-α, IL-6, IL-1ß), the reversal of apoptosis-related proteins expression (Bcl-1, Bax), the increase of cell viability and the decrease of the proportion of apoptotic cells. The result of Luciferase activity assay demonstrated that miR-543 directly targets Bcl-2. CONCLUSIONS: MiR-543 expression was increased in LPS-treated HK-2 cells, and silencing miR-543 could inhibit LPS-induced inflammation and apoptosis in HK-2 cells via targeting Bcl-2.

[Rare thalassemia caused by novel nucleotide variants in the globin gene: four case reports and literature review].

Objective: To analyze the DNA sequences and clinical phenotypes of four cases with rare thalassemia to improve its recognition and accurate diagnosis. Methods: The DNA sequence characteristics of four cases with rare thalassemia diagnosed from May 2014 to December 2019 were retrospectively analyzed, and related literature was reviewed. Results: The results of the routine gene test for thalassemia indicated that the common three type of deletion and three point mutations in hemoglobin alpha 1/2 (HBA1/A2) , and 16 point mutations in hemoglobin beta (HBB) gene were unable to be detected in cases 1-3, and case 4 was--SEA. However, the results of HBA1/A2 and HBB whole-genome sequencing revealed that the four cases had a point mutation of HBB:c.347C>A, HBB:c.1A>G, HBB:c.393T>G, and HBA2: c.301-1G>A (IVS II-142 G>A) , respectively. Meanwhile, the father, aunt, and grandfather of case 2 carried the HBB:c.1 A>G heterozygous point mutation. Conclusion: The novel mutations in HBB and HBA2 genes, resulting in a rare thalassemia, were revealed. Among them, the HBB:c.347C>A, HBB:c.1A>G, and HBA2:c.301-1G>A (IVS II-142 G>A) mutations were first reported in Chinese patients with thalassemia. Contrarily, HBB:c.393T>G mutation has not yet been recorded in the databases of human hemoglobin variants and thalassemia. The discovery of these novel nucleotide variants in this study would enrich the DNA mutation gene database of thalassemia.

Persistence of antibodies acquired by natural hepatitis E virus infection and effects of vaccination.

OBJECTIVE: Naturally acquired anti-hepatitis E virus (HEV) immunity can protect against new HEV infections. The aim of this study was to analyse the persistence of naturally acquired anti-HEV immunoglobulin (Ig) G and anti-HEV IgG concentrations after vaccination. METHODS: We examined the seropositivity rates of participants included in a phase 3 clinical efficacy trial (67 months' follow-up) for a HEV vaccine (Hecolin; Xiamen Innovax Biotech, China) and predicted long-term persistence using mixed-effect models. RESULTS: The analysis focused on 2242 baseline seropositive participants in a control group (placebo recipients) and 2031 baseline seropositive participants in an vaccine group (vaccine recipients) who received 1 to 3 doses of Hecolin. Naturally acquired anti-HEV IgG levels decreased steadily independent of the initial antibody level; 50% of the placebo recipients were expected to have undetectable antibody concentrations after 14.5 years. After immunization with Hecolin, the power-law model and the modified power-law model predicted that 82.1 and 99.4% of the participants, respectively, would remain seropositive for anti-HEV IgG for 30 years after vaccination. CONCLUSIONS: Whereas naturally acquired anti-HEV IgG levels decrease steadily, HEV vaccination induces long-lasting, high-level anti-HEV IgG concentrations.

Protection against hepatitis E virus infection by naturally acquired and vaccine-induced immunity.

Immunity acquired from infection or vaccination protects humans from symptomatic hepatitis E. However, whether the risk of hepatitis E virus (HEV) infection is reduced by the immunity remains unknown. To understand this issue, a cohort with 12 409 participants randomized to receive the hepatitis E vaccine Hecolin(®) or placebo were serologically followed up for 2 years after vaccination. About half (47%) of participants were initially seropositive. A total of 139 infection episodes, evidenced by four-fold or greater rise of anti-HEV level or positive seroconversion, occurred in participants who received three doses of treatment. Risk of infection was highest among the baseline seronegative placebo group participants (2.04%). Pre-existing immunity and vaccine-induced immunity lower the risk significantly, to 0.52% and 0.30%, respectively. In conclusion, both vaccine-induced and naturally acquired immunity can effectively protect against HEV infection.

Effects of Rhynchophylline on relaxation and contraction of the bladder detrusor in rats.

AIM: The aim of this study was to observe the effects of Rhynchophylline (Rhy) on the relaxation and contraction of rat bladder detrusor and urodynamics and determine the changes in the tension of isolated rat bladder muscle strips. MATERIALS AND METHODS: Rats were randomly divided into four groups: sham-operated, overactive bladder (OAB) model, Rhy-treated, and the control group. Sections of urodynamic testing and electrophysiological OAB indicators of detrusor were measured. The effect of tension on the isolated rat bladder detrusor muscle strips was determined; activators and antagonists of calcium-activated potassium ion channels were detected in vitro using the tension method. The contraction of detrusor muscle strips and the antagonism of acetylcholine due to changes in muscle contraction were observed. RESULTS: The Rhy-treated group significantly decreased the maximum bladder capacity, bladder filling pressure, leak point pressure, contraction frequency, motility index (p < 0.05). The affinity index of Rhy was 4.53 ± 0.22. However, 1 µmol/L to 2 µmol/L Rhy shifts CaCl2 cumulative dose-response curves to the right in a non-parallel manner, showing a non-competitive antagonism. Rhy inhibits detrusor contraction by blocking L-type calcium channels and activating big-conductance calcium-activated potassium channels. A low concentration of Rhy can inhibit muscle contraction caused by intracellular calcium. CONCLUSIONS: Rhy plays an important role in OAB treatment and decreases effectively on sections of urodynamic testing and electrophysiological OAB indicators of detrusor.

Inorganic phosphorus fractionation and its translocation dynamics in a low-P soil.

The translocation of different inorganic phosphorus (Pi) forms in a low-P soil (Langfang experimental station, Hebei province, China) over time was investigated using P fractionation extraction and a (32)P tracer technique. The L-value and P availability of the soil was assessed using 5 different maize genotype (Zea mays L.) cultivars. The results showed that the different Pi fractions in the soil increased in the order of H(2)SO(4)-extractable P (Ca(10)-P) > Na(3)C(6)H(5)O(7)-Na(2)S(2)O(4)-extractable P (O-P) > NH(4)Ac-extractable P (Ca(8)-P) > NaHCO(3)-extractable P (Ca(2)-P), NH(4)F-extractable P (Al-P), NaOH-Na(2)CO(3)-extractable P (Fe-P), and the content of plant-unavailable P (Ca(10)-P + O-P) was high, up to 79.1%, which might be an important reason for P deficiency in this low-P soil. The (32)P tracer results showed that after the addition of (32)P-Pi to the soil with no P fertilizer applied for 25 d, 29.0% of (32)P was quickly transformed into Ca(2)-P (rapidly available P), and 66.1% of (32)P was transformed into Al-P, Fe-P and Ca(8)-P (slowly available P). Only 5.0% of (32)P was transformed into O-P and Ca(10)-P (plant-unavailable P). Moreover, in the soil with P fertilizer applied, (32)P transformation into Ca(2)-P increased, and the transformation into Ca(8)-P + Fe-P + AL-P and O-P, Ca(10)-P significantly decreased compared to the soil with no P fertilizer applied (p < 0.05). This result suggested a higher rate for water-soluble P transformation to slowly available and plant-unavailable P in P deficient soil than in soil with sufficient P. The results of maize L-value determination showed that different genotype maize cultivars had different soil P-use efficiency and low-P tolerance mechanisms. Low-P tolerant cultivar DSY-32 regulated soil P-use efficiency and plant P content according to exogenous P fertilizer application. However, another low-P tolerant cultivar, DSY-2, used soil P more efficiently, regardless of the application of exogenous P.

Assessment of soil contamination with Cd, Pb and Zn and source identification in the area around the Huludao Zinc Plant.

The distribution characteristics of heavy metals (cadmium (Cd), lead (Pb) and zinc (Zn)) in the natural soil profiles around the Huludao Zinc Plant (HZP), an old industrial base in Northeast China, were analyzed. The pollutant source was identified using (210)Pb isotope technique to evaluate the geochemical characteristics of Pb and the historical production records of HZP. The results indicated: dust precipitation from HZP was the primary source of the pollutants. The average deposition rates of Cd, Pb and Zn were 0.33, 1.75, and 30.97 g/m(2)year, respectively at 1 km away after HZP, and 0.0048, 0.035, and 0.20 g/m(2) year, respectively at 10 km away after HZP. There is a risk of secondary pollution to the environment as well as the food chain in seriously polluted areas used for cultivation.

Effects of external phosphorus on the cell ultrastructure and the chlorophyll content of maize under cadmium and zinc stress.

Using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analysis, it was found that the ultrastructure of chloroplasts were changed, the shape of the chloroplasts altered and the numbers of grana that were asymmetrical increased; the numbers of grana and thylakoids decreased under the stress of Cd and Zn. The results indicated that the complex pollution involving Cd and Zn resulted in the membrane system of chloroplasts being damaged. When external phosphorus was applied, the numbers of damaged chloroplasts were significantly reduced and the nucleoli were better formed than those that did not receive phosphorus treatment. Moreover, many phosphate deposits were found in the vacuoles and on the surface of the roots, which were formed by phosphorus complexing with Cd (K(sp)=2.53x10(-33)) and Zn (K(sp)=9.00x10(-33)), respectively. Treatment with phosphorus conduced an increased chlorophyll content in plants compared with those that did not receive external phosphorus.

[Listeria monocytogenes induces thymocyte apoptosis in mice].

The murine thymocyte apoptosis induced by Listeria monocytogenes(LM) was detected with morphology, FCM, and DNA electrophoresis. The results were that LM elicited typical morphological changes of thymocyte apoptosis; the typical apoptosis peak was displayed with FCM, and typical "ladder pattern" with agarose gel electrophoresis. The apoptotic cells were found at 8 h after the mice had infected LM and reached climax at 48 h. The thymus weight significantly reduced at 16 h, and reached the lowest at 48 h after the mice had infected LM. The percentage of apoptotic cells was raised with the increasing of LM. These results suggest that LM induces thymocyte apoptosis in dose- and time-dependent manner.

[Experimental study on isolated testes with ischemia/reperfusion injury].

OBJECTIVE: To investigate the injury on isolated testes induced by ischemia/reperfusion(I/R), and the protective effect of Yisheng injection on the injury. METHODS: Twenty-six isolated cadaver testes contributed by 13 persons were preserved with 4 degrees C 250 ml hypertonic citrate alloxuric (HCA) solution and then reperfused with 37 degrees C 500 ml HCA. Solution of experimental group contained 500 micrograms/ml Yisheng injection. In simple cold preservation test, involving in 8 experimental and 8 control testes, a series of time points (6, 12, 18, 24, 36, 48, 60, 72 hours) were set to harvest. 10 testes (1 testis respectively on 6, 12, 18, 24 and 36 hours in experimental and control groups) were reperfused with 37 degrees C HCA for 6 and 12 hours. Histological and histochemical changes were observed. RESULTS: In the experimental testes, 4 degrees C cold preservation in 24 hours could not induce obvious pathologic changes. After 24 hours, changes such as swelling, vacuolar degeneration or detachment of endothelial cells (ECs), separation between basement membrane and seminiferous epithelium, mal-alignment of spermatogenous cell and edema of mesenchyme could be observed. In the testes preserved for 12 hours, the activity of lactic dehydrogenase(LDH) and succinic dehydrogenase (SDH) increased, then fallen after 24 hours. The activity of Nitric oxide synthetase(NOS) decreased after 18 hours. All changes were more obvious after following 37 degrees C reperfusion. In the control testes, swelling and vacuolar degeneration of ECs occurred on 12 hours cold preservation, and injury was worse along with the prolongation of cold preservation time. Pathologic changes of ECs, seminiferous epithelium and mesenchyme were serious after 37 degrees C reperfusion. CONCLUSION: 4 degrees C cold preservation in 24 hours can only cause mild ECs' injury, and obvious abnormal testes' histological profile can be observed beyond 24 hours. 37 degrees C reperfusion will make injury worse. Yisheng injection can keep isolated testes histologic structure well in 24 hours cold preservation, and it has protective effect on I/R injury.

[Intravenous infection of C. albicans induces apoptosis of cortical thymocytes in mice].

In this study, we aimed to understand the location of apoptotic cells in murine thymus and phenotype of murine thymocytes involved in C. albicans induced apoptosis. Results showed that significant increase of apoptotic cells in thymus began at 6 h after intravenous injection of C. albicans and kept increased at 12 h, 18 h and 24 h. Quantitative changes of thymocytes with different phenotype measured by flow cytometry revealed that percentage of CD3+ and CD8+ cells were significantly reduced compared to the controls. Moreover in situ cell death detection of thymic tissue revealed that apoptotic cells induced by C. albicans mainly located in the thymic cortex, while in the thymic medulla a very small number of thymocytes was involved. Taken together our study indicates that C. albicans induces apoptosis of murine thymocytes, moreover the apoptosis possibly involves the major immature CD3+ CD4+ CD8+ cells in the thymic cortex and minor mature CD3+ CD4- CD8+ cells in the thymic medulla.

[Clinical application of inspiring psychological nursing in vaginal delivery].

A new nursing method named Inspiring Psychological Nursing was initiated and applied to 176 primigravida undergoing vaginal delivery from Jan. to Jun. 1994. 202 primigravida who received general medical care during vaginal delivery from Jan. to Jun. 1993 was served as control group. No significant differences were found between the two groups regarding to age, gestational weeks, times of pregnancy, fetal position and newborn's weight. However significant differences were found regarding the duration of the second stage of labor and incidence of newborn asphyxia. Primigravida in the control group usually delivered after 60 min. and carried a higher incidence of newborn asphyxia, being 6.7%. It's concluded that Inspiring Psychological Nursing could significantly improve pregnancy outcome.

[Significance of measurement of serum and cerebrospinal fluid trace elements in the diagnosis of brain tumors].

Cu, Zn, Cd, Mn, Se in serum and Cu, Cd and Mn in cerebrospinal fluid (CSF) of 150 patients were measured by flame, flameless and hydride generation atomic absorption spectrophotometry. The patients were divided into three groups: malignant brain tumor group (MTG), benign brain tumor group (BTG), and non-neoplastic neurological disease group (no evidence of brain tumor group, NENG). The results were as follows: 1. Serum Cu, Zn, Cd, Mn, Se levels before operation were higher than those after operation; 2. Cu Cd, Mn serum levels were positively correlated with the CSF in the three groups. The correlation coefficients of serum Cu-CSFCU were 0.8935 (MTG), 0.6074 (BTG), 0.5349 NETG); serum Cd-CSFCd 0.7963, 0.2671, 0.5398; serum Mn- CSF Mn 0.5855, 0.4474, 0.5163, (P less than 0.01). 3. Serum and CSF Cu levels in MTG were significantly higher than in BTG and NETG (P less than 0.01). 4. A group of discrimination function equations was established with stepwise discrimination analysis. The six factors, i. e. serum Cu, Cd, Mn, Se and CSF Cu, Mn were significant in the equations. The conformation rate of diagnosis and discrimination by the equations in the malignant and benign brain tumors with clinical diagnosis was 81 percent. The equations could describe the relationship of coordination and antagonism among trace elements in serum and CSF. The authors believe that discrimination equations may have potential use both in diagnosing and discriminating patients in the 3 groups mentioned above and in following patients after excision of the brain tumor.

[The clinical significance of multifactor discrimination and analysis of maternal serum Cu, Zn, Cd, Mn contents in the diagnosis of abnormal fetus].

In this paper we present pair-comparison test with serum contents of Cu, Zn, Cd, Mn, Se in pregnant women with neural tube defects fetus (NTD), pregnant women with normal fetus and healthy with no pregnancy using Bayes Stepwise Discrimination Analysis and Fisher Discrimination Analysis. The results showed: NTD of fetus was related to deficiency of maternal serum in Cu, Zn, Mn and to excess of Cd. The maternal serum levels of Cu, Cd and Mn can be used as sensitive indicator to detect intrauterine abnormality at an early stage. The low levels of maternal serum in Zn and Se were not a specific indicator of NTD but a characteristic shared by all pregnant women. It is important to pay attention to supplying pregnant women at an early stage with Zn, Se to meet the needs of normal growth of the fetus and prevent foetal abnormality. With stepwise discrimination we established two equations by Cu, Zn, Cd, Mn used for distinguishing normal from abnormal fetus. The accordant rate of clinical discrimination with classification output for inside examples was 93%. In the equation established by Fisher principle Mn had the greatest rate, namely, 37.6%.

Clinical significance of hair cadmium content in the diagnosis of mental retardation of children.

Cadmium (Cd) is known to be closely related to the development of children's intelligence. To investigate the clinical significance of the hair cadmium content in the diagnosis of mental retardation (MR), 415 normal and 85 MR children's hair cadmium contents were measured by 79-1 anodic stripping voltmeter. The data were processed by an Apple-II microcomputer. Comparing the median value and using the Chi-square test the Cd contents in the hair of both male and female MR children were obviously higher than those of normal children (P less than 0.001). If the Cd content in the hair of male children is higher than 0.239 microgram/g, and higher than 0.180 microgram/g in the female, MR can be diagnosed. The sensitivity and specificity of the test are greater than 90%; false positive and false negative rates are lower than 10%. Hair Cd can be used as an additional criterion in the diagnosis of MR and may be important in the screening for MR.