Skimmed Milk Diluent Promotes the Sperm Motility and Conception Rate of Dorper Sheep Compared to Vitamin B12 Diluent.

BACKGROUND: Dorper sheep is an ideal breed for improvement, with higher meat production and increased adaptability. Artificial insemination is an efficient technique for Dorper genetic improvement and reproduction management. However, there is no uniform diluent for Dorper semen dilution. OBJECTIVE: To compare the effects of vitamin B12 (VB12) and skimmed milk diluents on sperm motility at different ratios and time points, and the effects on conception rate. MATERIALS AND METHODS: We detected the effect of diluents on sperm density, deformity, motility and conception rate of Dorper sheep. RESULTS: We found the optimal dilution ratio of skimmed milk is 1:3. Compared to VB12, skimmed milk at 1:3 ratio prolonged semen storage time (48 h vs. 18 h, storage at a low temperature of 4°C) and increased the survival index of sperm (44.7 ± 2.8 vs. 18.5 ± 0.6, P<0.01). CONCLUSION: Skimmed milk is more effective, nutritious and convenient than vitamin B12, representing a more advantageous diluent.

Secondary acute angle closure attack as an initial presentation of a novel type of systemic lupus erythematosus.

BACKGROUND: We report a rare case of secondary acute angle closure attack because of lupus choroidopathy and accompanying polyserositis, as an initial presentation of a novel type of systemic lupus erythematosus in a 44-year-old woman. CASE PRESENTATION: The patient complained of eyelid oedema and chemosis with bilateral severe loss of visual acuity. Systemic lupus erythematosus was diagnosed based on malar rash, polyserositis, proteinuria and positive antibody titers for antinuclear antibodies, anti-DNA, antinucleosome antibodies and ribosomal RNP. Subsequently, secondary bilateral acute angle closure caused by choroidal effusions with lupus choroidopathy was diagnosed. A month after steroid and immunosuppressive drug therapy, the patient's intraocular pressure and visual acuity returned to normal. During the subsequent year, the secondary acute angle closure did not recur and polyserositis remained under control. CONCLUSIONS: Bilateral, secondary acute angle closure attack due to SLE choroidopathy can be an initial presentation of SLE, which is often accompanied by polyserositis. Prompt and aggressive high doses of steroids and immunosuppressive therapy are strongly recommended.

[Abnormal functional connectivity between sublaterodorsal nucleus and the motor cortex in Parkinson's disease with rapid-eye-movement sleep behavior disorder].

Objective: To investigate the abnormal functional connectivity between sublaterodorsal nucleus (SLD) and the brain motor areas in Parkinson's disease with or without rapid-eye-movement (REM) sleep behavior disorder by resting state functional magnetic resonance imaging (rfMRI). Methods: A total of 64 subjects recruited in the Department of Neurology, the People's Hospital of Zhengzhou University were enrolled in this study from August 2014 to December 2017 according to international diagnosis criteria, 20 cases with PD-RBD (PD-RBD group), 23 cases without PD-nRBD (PD-nRBD group) and 21 age, gender-matched healthy controls (HC group). All subjects were examined by Hoehn-Yahr, UPDRS-Ⅲ and rfMRI. Resluts: UPDRS-Ⅲ scores and Hoehn-Yahr staging were showed significantly different from that in the HC group(KW=-27.35, P<0.001) but showed no significantly difference from that in the PD-nRBD group(KW=6.01, P=0.807). Compared with the control group, the FC analysis showed reduced correlations from the left SLD to the right orbital middle frontal gyrus(T=-4.567 8, P<0.001), the left cingulate gyrus(T=-3.196 0,P<0.001), the left cerebellum(T=-4.267 0,P<0.001) and the right midbrain(T=-4.773 7, P<0.001), from the right SLD to the bilateral cerebellum (T(left)=-5.040 1, T(right)=-4.210 5, P<0.001), the left precuneus(T=-4.468 2,P<0.001) and the left precentral gyrus (T=-4.473 9,P<0.001) in the PD-RBD. The correlations between the left SLD and left cerebellum, right midbrain and between the right SLD and left precuneus, left precentral gyrus were negative correlated with the motor functional tests. Conclusions: There are abnormal functional connectivity from the SLD to the motor areas in PD-RBD patients, leading to clinical PD-RBD motor symptoms and movement disorders.

An immortalized microglial cell line (Mocha) derived from rat cochlea.

Microglia are glial-immune cells that are essential for the function and survival of the central nervous system. Microglia not only protect neural tissues from immunological insults, but also play a critical role in neural development and repair. However, little is known about the biology of microglia in the cochlea, the auditory portion of the inner ear. In this study, we detected TMEM119+, CD11b+, CD45+ and Iba1+ populations of cells in the rat cochlea, particularly in Rosenthal's canal, inner sulcus and stria vascularis. Next, we isolated and enriched the population of CD11b+ cells from the cochlea and immortalized these cells with the 12S E1A gene of adenovirus in a replication-incompetent retroviral vector to derive a novel microglial cell line, designated Mocha (microglia of the cochlea). The resulting Mocha cells express a number of markers consistent with microglia and respond to lipopolysaccharide (LPS) stimulation by upregulation of genes (Cox2, ICAM-1, Il6r, Ccl2, Il13Ra and Il15Ra) as well as releasing cytokines (IL-1beta, IL-12, IL-13 and RANTES). As evidence of microglial function, Mocha cells phagocytose fluorescent beads at 37°C, but not at 4°C. The expression pattern of microglial markers in Mocha cells suggests that immortalization leads to a more primitive phenotype, a common phenomenon in immortalized cell lines. In summary, Mocha cells display key characteristics of microglia and are now available as a useful model system for the study of cochlear microglial behavior, both in vitro and in vivo.

Effects of beta 2 adrenergic agonists on axonal injury and mitochondrial metabolism in experimental autoimmune encephalomyelitis rats.

The primary aims of this study were to investigate mitochondrial metabolism during experimental allergic encephalomyelitis (EAE) animal model axonal injury and to determine the correlation among neurological function scores, pathological changes, and the activities of the BB isoenzyme of creatine kinase (CK-BB), catalase (CAT), and calpain in the brain tissues of EAE rats. Another goal was to preliminarily define the mechanism of mitochondrial metabolism resulting from the effect of beta 2 adrenergic agonists in the process of EAE animal model axonal damage. EAE was induced in specific pathogen free Wistar rats by guinea pig spinal cord homogenate, complete Freund's adjuvant, and pertussis vaccine. We recorded the behavioral change in EAE rats, detected pathological changes in central nervous tissue, and observed the changes of the CK-BB, CAT, and calpain in the EAE rat brain and spinal cord. The results indicated that the average neurologic function score increased in the EAE group compared to that of the controls (P < 0.01). In addition, CAT and CK-BB activities significantly decreased and the calpain activity significantly increased compared with those of the control group (P < 0.05). The decrease of the activity of central nervous CK-BB and CAT content, as well as the increase of calpain activity at the highest time point were considered to be the consequences of EAE. Furthermore, the results revealed that use of salbutamol could alleviate disease symptoms and reduce the recurrence of the EAE disease.

Combining the triglyceride-glucose index and glycated hemoglobin A1c to assess the risk of preeclampsia in women with normal glucose tolerance: a cross-sectional study.

OBJECTIVE: This study aimed to explore the relationship between the triglyceride-glucose (TyG) index, glycated hemoglobin A1c (HbA1c), and preeclampsia in pregnant women without gestational diabetes mellitus (GDM). PATIENTS AND METHODS: This retrospective study included pregnancies with normal oral glucose tolerance tests (OGTTs) from March 2018 to February 2019. During the second trimester, serum lipids, fasting plasma glucose (FPG), and HbA1c were measured, and OGTTs were performed. Participants were classified into four groups based on their TyG index and HbA1c levels. Logistic regression analysis was done to determine the odds ratios (ORs), and receiver operating characteristic (ROC) curve analysis was used to evaluate the ability of the TyG index and HbA1c to predict the risks of preeclampsia. RESULTS: Patients with preeclampsia exhibited higher TyG index and HbA1c levels (all p < 0.001). The incidence of preeclampsia increased with elevated TyG index and HbA1c levels individually. Furthermore, the highest incidence of preeclampsia was observed when both the TyG index and HbA1c levels were elevated. ROC curve analysis revealed that the combined TyG index and HbA1c displayed an area under the curve (AUC) of 0.689 in predicting the risk of preeclampsia. Even after adjusting for potential confounding factors, the risk of developing preeclampsia remained significantly higher. These associations were especially prominent in women aged ≥ 35 years or those with a normal BMI. CONCLUSIONS: The findings of this study indicate that increased TyG index and HbA1c levels are associated with a higher incidence and risk of preeclampsia in women with normal glucose tolerance during pregnancy. The TyG index and HbA1c levels may serve as potential markers for preeclampsia in individuals with normal OGTT results.

Reference intervals of systemic immune-inflammation index, neutrophil-to-lymphocyte ratio, lymphocyte-to-monocyte ratio, and platelet-to-lymphocyte ratio during normal pregnancy in China.

OBJECTIVE: To observe the changes in systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR) during normal pregnancy and establish appropriate reference intervals (RIs) for healthy pregnant women. PATIENTS AND METHODS: This retrospective study was conducted from March 2018 to February 2019. Blood samples were collected from healthy pregnant and nonpregnant women. The complete blood count (CBC) parameters were measured, and SII, NLR, LMR, and PLR were calculated. RIs were established using the 2.5th and 97.5th percentile of the distribution. Besides, the differences in CBC parameters between three pregnant trimesters and maternal ages were also compared to assess their influences on each indicator. RESULTS: SII and NLR in three pregnant trimesters increased in pregnant women, and the upper limit of SII and NLR in trimester 2 showed the highest value. On the contrary, LMR decreased in all three pregnant trimesters compared with nonpregnant women, and the values of LMR and PLR showed a gradual downward trend along with the trimesters. Besides, RIs of SII, NLR, LMR, and PLR during different trimesters in different age partitions showed that the values of SII, NLR, and PLR increased with age in a general trend, while LMR showed the opposite trend (p < 0.05). CONCLUSIONS: The SII, NLR, LMR, and PLR showed dynamic changes during pregnant trimesters. RIs of SII, NLR, LMR, and PLR for healthy pregnant women according to pregnant trimesters and maternal age were established and validated in this study, which will promote the standardization of clinical application.

Lactobacillus coryniformis subsp. torquens inhibits bone loss in obese mice via modification of the gut microbiota.

High-fat diet (HFD)-induced obesity results in bone loss associated with an imbalanced gut microbiota and altered immune status. Probiotics are live microorganisms that are beneficial to the host and are important in maintaining bone health and gut homeostasis. In this study, the probiotic Lactobacillus coryniformis subsp. torquens (T3L) was isolated from traditional yak milk cheese produced in Lhasa and showed distinct acid and bile salt resistance as potential probiotics. Our data indicated that T3L not only reversed HFD-induced gut dysbiosis, as indicated by decreased Firmicutes-to-Bacteroidetes ratios but also reduced bone loss. The anti-obesity, microbiome-modulating, and bone-protective effects were transmissible via horizontal faeces transfer from T3L-treated mice to HFD-fed mice. The protective effects of T3L on bone mass were associated with regulatory T (Treg) cell-mediated inhibition of osteoclast differentiation. Our data indicate that T3L is a regulator of the gut microbiota and bone homeostasis in an animal model.

Long non-coding RNA FAM201A promotes lung squamous cell carcinoma progression through interaction with miR-101.

OBJECTIVE: This study aimed to investigate the mechanism of LncRNA FAM201A mediating lung squamous cell carcinoma progression through interaction with miR-101. PATIENTS AND METHODS: NCI-H520 cells and SK-MES-1 cells were transfected with miRNA-101-mimics and miRNA-101-inhibitor, the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect FAM201A and miR-101 expression. CCK-8, Wound healing assay and transwell assay were utilized to detect the influence of FAM201A on the malignancy of NCI-H520NCI-H520 and SK-MES-1SK-MES-1 cells. Cell apoptosis was determined by flow cytometry. The underlying pathways of FAM201A were measured using Western blot. Xenograft tumor experiments were conducted to detect tumor growth and metastasis in vivo.NCI-H520SK-MES-1 Kaplan-Meier method calculated patient survival. RESULTS: (1) Silencing of FAM201A inhibited the proliferation, migration and invasion of NCI-H520 and SK-MES-1cells and stimulated cell apoptosis significantly. Furthermore, FAM201A elimination hindered tumor growth and metastasis in vivo. (2) Compared with the si-control group, the protein expression of Ki67, Vimentin, Cleaved-caspase-3 and N-cadherin were decreased in the si-FAM201A group. (3) After transfection of miR-101-mimics, the expression level of Vimentin protein was significantly increased, while the expression level of Vimentin protein was significantly decreased after miR-101-inhibitor transfection. (4) MiR-101 mimics could alleviate FAM201A silencing-induced inhibitive effects on cell proliferation, migration, invasion and promotive effects on cell apoptosis. CONCLUSIONS: FAM201A could target miR-101 and upregulate Vimentin to inhibit lung cancer progression. FAM201A was expected to be a potential biomarker and therapeutic target for lung cancer.

Injection of Anti-proBDNF in Anterior Cingulate Cortex (ACC) Reverses Chronic Stress-Induced Adverse Mood Behaviors in Mice.

Foraging behavior is a species-specific behavior which is considered to involve the decision making and higher cognitive functions. We previously established a novel method to detect the foraging behavior in chronic unpredictable mild stress (CUMS)-induced depression mice, in which the food foraging activity of mice was significantly reduced. Furthermore, it is generally assumed that the bilateral anterior cingulate cortex (ACC) is related to foraging activity in rat. Brain-derived neurotrophic factor (BDNF) is widely expressed in many regions of the brain and is down-regulated in depressive patients. However, the relationship between the precursor of brain-derived neurotrophic factor (proBDNF) and depression has not been fully elucidated. The results showed that CUMS in mice induced anxiety- and depression-like behaviors and significant reduction in BDNF messenger RNA (mRNA) in the brain. In this study, we evaluated the effect of anti-BDNF and anti-proBDNF in the ACC on the CUMS-induced depression mice. In contrast to the normal IgG group (normal IgG microinjection into the ACC), bilateral ACC treatment with anti-proBDNF microinjection not only reversed depressive activity but also significantly increased the amount of foraged food and BDNF mRNA in the brain. There was no significant alteration in the group of anti-BDNF microinjection into the ACC. Our data indicate that the proBDNF signaling pathway might down-regulate the foraging activity in CUMS rodents and be involved in the depression.

The predictive value of baseline LDL-TG level on major adverse cardiovascular events in a followed up cohort population.

OBJECTIVE: We aimed at identifying the predictive roles of Low-Density Lipoprotein Triglycerides (LDL-TG) for major adverse cardiovascular events (MACEs). PATIENTS AND METHODS: A longitudinal study in a routine health check-up population was performed with an average follow-up of 4.8 years. The participants involved in this study were 1680, from 2007 to 2009, and all had followed-up for all-cause mortality, cardiovascular disease mortality, and the development of MACEs. The demographic information and anthropometric parameters at baseline were recorded. The baseline and follow-up conventional lipid parameters were measured. We also examined the level LDL-TG, as well as the relationship between its level and MACEs. RESULTS: MACEs individuals were characterized by statistically higher baseline LDL-TG (17.22 ± 8.05 vs. 16.39 ±7.35 nmol/l, p = 0.017). The univariate regression for MACEs group indicated that the LDL-TG (b = 0.813, HR = 2.254, 95% CI: 1.454-3.494, p < 0.001), older age, sex and other factors were a significant risk for MACEs. Furthermore, in the adjusted Cox model showed that only higher baseline LDL-TG (b =0.512, HR = 1.669, 95% CI: 1.013-2.748, p = 0.044) and older age (b = 0.062, HR = 1.064, 95% CI: 1.034-1.094, p < 0.001, Table IV) were still predictors for MACEs. CONCLUSIONS: Higher baseline LDL-TG closely associated with MACEs and it is a moderate and independent predictive factor for MACEs.

Enhanced aggressive behaviour in a mouse model of depression.

Depression is one of the most common chronic mental disorders, which is a leading cause of morbidity and mortality in patients. Depression often leads to offensive and defensive behaviours but the underlying mechanisms are not known. We propose that the aggressive behaviours in depression can be modelled in animal experiments. In this study, we successfully established a mouse model of depression using the chronic unpredictable mild stress (CUMS) paradigm and detected aggressive and social dominance behaviours in rodents by resident/intruder test and social dominance tube test (SDTT), respectively. The CUMS-exposed mice showed increased defensive, offensive and aggressive behaviours in the resident-intruder test. In the SDTT, these mice showed enhanced social dominance. These alterations were associated with reduced MAP-2 expression in the hippocampus while no difference in ß-tubulin expression was detected. In addition, the treatment of anti-depressant fluoxetine reversed the aggressive behaviours without reducing the social dominance behaviour induced by CUMS. However, fluoxetine did effectively reverted the changes in MAP-2 expression in the hippocampus. In addition, the nonspecific tricyclic antipsychotic drug, clozapine, reversed all symptoms of CUMS-exposed mice including aggressive tendencies, impulsive violence, social dominance behaviour and MAP-2 expression in the hippocampus. The results suggests that social maladjustment such as competition and social dominance are likely related to the dopaminergic system rather than the serotonergic system and the hippocampal dendritic structure protein MAP-2. Thus, dominance can be separated from aggression. This study shows that aggression/hostility and social hierarchy/dominance are increased in the CUMS-exposed mice and thus provide an excellent model for further study in the diagnosis and the treatment of depression-associated aggression.

Sequence of an operon containing a novel delta-endotoxin gene from Bacillus thuringiensis.

An insect larval toxin designated CryII is produced by several subspecies of Bacillus thuringiensis and differs from the other major delta-endotoxins in these bacteria in its size, toxicity profile and presence as part of an operon with three open reading frames (ORF). Such an operon from a novel B. thuringiensis isolate has been cloned and differs from one previously characterized in the following ways: (a) the size and number of amino acid repeats in one of the ORFs; (b) the smaller size of the CryII protoxin and the presence of a unique 110-kDa CryII-related antigen; and (c) high larvicidal activity for a particular Lepidopteran but low activity for a Dipteran. Various subclones of this operon were introduced into a plasmid-free B. thuringiensis strain and only the cryII gene was found to be necessary for protoxin accumulation.

[Insect-resistant transgenic plants of Brassica napus and analysis of resistance in the plants].

Cotyledons, each with a 1-2 mm petiole at its base, were cut from axenic seedlings and infected with Agrobacterium tumefaciens. After 2-3 days of cocultivation, the cotyledon explants were transferred to MS selection medium containing 15 mg/L kanamycin and 4.5 mg/L 6-BA to induce shoot differentiation. Kanamycin-resistant shoots were subcultured on selection medium with 20-50 mg/L kanamycin for 3-6 months for eliminating escaped non-transformants, and then rooted on MS medium containing 25 mg/L kanamycin and 0.1 mg/L NAA. Whole plants were transplanted into soil and grew in the field. DNA Southern blot hybridization and polymerase chain reaction showed that some of the plants were positive when probed with the insecticidal crystal protein gene. The transgenic plants exhibited tolerant to pest insects such as Laphygma exigua and Pieris rapae in leaf feeding experiments Kanamycin-resistance and insect-resistance were maintained in the progeny. The foreign genes were delivered to the progeny according to Mendelian Law of single gene segregation.

Foraging activity is reduced in a mouse model of depression.

Depression interferes with the human ability to make decisions. Multiple criteria have been adopted for the diagnosis of depression in humans, but no clear indicators are available in animal models to reflect the depressive mood, involving higher cognitive functions. The act of foraging is a species-specific behaviour which is believed to involve the decision-making and higher cognitive functions. We previously established a method to detect the foraging behaviour of rodents, in which our results demonstrated that NMDA and dopamine receptors were involved. Conversely, increased NMDA receptors and reduced dopamine have been reported in depression model rodents. However, we hypothesise that foraging activities may also be impaired in depression. To test the theory, we successfully established a mouse model of depression using the chronic unpredictable mild stress (CUMS) paradigm. Most interestingly, the food foraging activity of mice after CUMS was significantly reduced. In addition, the treatment of anti-depressant fluoxetine reversed most depressive symptoms and reduced glial fibrillary associated protein (GFAP) expression in the hippocampus, but was less effective in the reduction of foraging activities. However, clozapine reversed all symptoms of CUMS-exposed mice including reduction of GFAP expression in the hippocampus and impaired foraging activity. Our findings of GFAP expression as a marker to validate the CUMS protocol provide further validation of our hypothesis, that the reduced food foraging is probably a new behavioural finding of depression in which the serotoninergic system could not be singly involved. Our study suggests that NMDA receptors, serotoninergic and dopaminergic systems are differentially involved in these food foraging behaviours. Our data suggest that the foraging test in rodents can be a useful tool to assess the ability of decision-making in depression.

Effects of transgenic rice expressing Bacillus thuringiensis Cry1Ab protein on ground-dwelling collembolan community in postharvest seasons.

During 2005-2008, field studies were conducted at two locations in Chongqing, China, to assess the potential effects of transgenic rice expressing Bacillus thuringiensis (Bt) Cry1Ab protein on the nontarget ground-dwelling collembolan community in three postharvest seasons. Collembolans in non-Bt and Bt rice fields were sampled with pitfall traps during each of two postharvest seasons of 2005/2006 and 2006/2007 and litterbag traps during each of three postharvest seasons of 2005/2006, 2006/2007, and 2007/2008. Ground-dwelling collembolans in rice fields during the postharvest seasons were abundant, whereas community densities varied considerably between the two locations and among the three seasons. A total of 67,310 collembolans, representing three species, Entomobrya griseoolivata, Hypogastrura matura, and Bourletiella christianseni, were captured during the three postharvest seasons. E. griseoolivata was the predominant species, accounting for 87.7% of the total captures, followed by H. matura (10.7%) and B. christianseni (1.6%). In general, there were no significant differences in species compositions and abundances of each species between Bt and non-Bt paddy fields, suggesting no significantly impact of plant residues of Cry1Ab rice on collembolan communities during postharvest seasons.

Transgenic plants of rutabaga (Brassica napobrassica) tolerant to pest insects.

Cotyledons cut from axenic seedlings were immersed inAgrobacterium tumefaciens suspension which was treated with acetosyringone and nopaline at low pH overnight. The infected cotyledon explants were cultured on MSB medium (MS salts + B5 Vitamins) containing 6-BA 3mg/1 for 2-3 days, and transferred onto selective medium (MSB with kanamycin 50-100 mg/l). Kanamycin-resistant shoots were selected. More than 60 regenerated plants were obtained. About 60% of the plants showed high NPT II activity. Southern blot hybridization showed that some of the plants gave a positive signal with the insecticidal crystal protein gene (cry IA gene) probe, and exhibited tolerant to insects such asPieris rapae (cabbage caterpillar) in leaf feeding experiments. Kanamycin-resistance and insect-resistance were maintained in the progeny.

[Enhanced resistance of transgenic tobacco expressing Shiva A gene against bacterial wilt disease (Pseudomonas solanacearum pv tabaci)].

Shiva A gene was introduced into tobacco mediated by Agrobacterium tumefaciens. Transgenic plants show enhanced resistance against bacterial wilt disease (Pseudomonas solanacearum pv tabaci). Compared with control plants, the disease indexes of transgenic tobacco plants Sc-2 and Sc-6 drop about 42.1% and 60.6%.