Rehmannia Glutinosa Polysaccharide Regulates Bone Marrow Microenvironment via HIF-1α/NF-κB Signaling Pathway in Aplastic Anemia Mice.

Aplastic anemia (AA), a rare disorder, is associated with bone marrow microenvironment (BMM). Presently, AA treatment is of great difficulty. This study aimed to explore the mechanism of action of Rehmannia glutinosa polysaccharide (RGP) in AA. Busulfan was used to induce AA in BALB/c mice; blood cell count and Ray's Giemsa staining were used to assess the severity of hematopoietic failure; HE was performed to assess the pathological state of the marrow cavity; ELISA was performed to assess IL-4, IL-10, IL-6, IL-12, IL-1ß, TNF-α, MCP-1, VEGF, and EPO; and WB was performed to evaluate the effects of RGP on the HIF-1α/NF-κB signaling. Significant downregulation of hemocyte levels in the blood and nucleated cells in the bone marrow was reversed by RGP and Cyclosporine A (CA). Compared with the AA group, dilating blood sinusoids, inflammation, hematopoiesis, decreased bone marrow cells and megakaryocytes were alleviated by RGP and CA, and the HIF-1α/NF-κB signaling was inhibited too. Notably, RGP was more effective when used in combination with CA. In this study, we established a relationship between BMM and the HIF-1α/NF-κB signaling pathway and found that RGP regulates BMM by suppressing the activation of the HIF-1α/NF-κB signaling. Thus, RGP exerts a pharmacological effect on AA.

Effect of Sunflower Kernel Peptides Produced by Dual-Degradation on the Growth Performance, Nutrient Digestibility, and Health Status of Broilers

The purpose of this study was to investigate the effect of sunflower kernel peptides produced by enzymatic digestion, fermentation, or both on the growth performance, nutrient digestibility, and health status of broilers. Four diets contained 20% of sunflower kernel meal as its raw form (CON) or degraded by protease (ESM), Bacillus pumilus (FSM), or both (DSM). A total of 480 yellow broilers at one day old were randomly distributed to 4 groups with 6 replicates of 20 chicks each. The feeding trial lasted for 63 d. Results showed that peptide content was increased (p<0.001) from 3.97% (CON) to 32.5% (ESM), 24.2% (FSM), and 39.1% (DSM). The three sunflower peptide groups improved (p≤0.001) feed intake and body weight gain. The peptide groups increased (p≤0.015) ileal apparent digestibility of dry matter, energy, crude protein, and amino acids (methionine, lysine, tryptophan, and threonine). Furthermore, the peptide groups improved (p≤0.029) the health status by increasing serum immunoglobulins (IgA, IgG) and glutathione peroxidase. Additionally, among the peptide groups, DSM showed more pronounced effects (p<0.05) on these parameters than ESM or FSM. It is concluded that dual-degradation by enzymolysis and fermentation has a better improvement in the nutrition and application of sunflower kernel meal in broilers.(AU)

Effect of Dietary Tetramethylpyrazine on Egg Production, Nutrient Retention and Cecal Bacterial Diversity in Aged Laying Hens

This study aimed to investigate the effect of tetramethylpyrazine (TMP) supplementation on egg production, nutrient retention and cecal microbiota diversity using 288 commercial Hy-Line brown hens as of wk 75 to 86. Four treatments consisted of TMP addition at 0 (control, basal diet), 100, 150 and 200 mg/kg of diet. The results showed that diets supplemented with TMP addition improved egg-laying rate as of wk 77 compared to the control, which led to an increase (p<0.001) of egg mass by 97-225 g/hen throughout the whole trial, and a linear increase (p=0.003) of egg mass to the incremental TMP doses was found. At wk 86, the apparent digestibilities of dry matter and crude protein were enhanced (p<0.05), exhibiting consistent linear increases (p≤0.033) with the TMP doses. However, TMP did not cause alpha and beta diversity of cecal microbiota. The results suggest that TMP can be an additive to improve egg production and nutrient digestibility of aged laying hens.(AU)

Location and Vascular Classification of 188 parathyroid glands in New Zealand White Rabbits / [Localização e Classificação Vascular de 188 glândulas paratireoidianas em coelhos brancos da Nova Zelândia]

The function and protection of the parathyroid glands are increasingly popular research topics. New Zealand white rabbits are the most commonly used animal model of parathyroid ischemia. However, information on the vasculature of their parathyroid glands is limited. We used 94 healthy New Zealand white rabbits, 3-4 months of age and 2-3kg in weight, for exploration of the parathyroid glands, which were stained using hematoxylin and eosin (HE) after removal. The following types were classified according to the relationship between the position of the inferior parathyroid gland and the thyroid: Type A, Close Type, Type B, and Distant Type. There were 188 cases, 4 where the inferior parathyroid glands were located near the dorsal side of thyroid (2.13%), 8 where the inferior parathyroid glands were located superior to the upper pole of the thyroid (4.26%), 20 where the inferior parathyroid glands were located parallel to the thyroid (10.64%), and 155 cases where the inferior parathyroid glands were located inferior to the lower pole of thyroid (82.45%). Identifying the location and classifying the vasculature of the parathyroid glands in New Zealand white rabbits will provide an anatomical model to assist in future research.(AU)

A função e proteção das glândulas paratireoidianas é um tópico de pesquisa cada vez mais popular. Coelhos brancos da Nova Zelândia são o modelo animal mais comumente usada para isquemia da paratireóide. Porém, informação sobre a vasculatura de suas glândulas paratireóides é limitada. Foram usados 94 coelhos brancos da Nova Zelândia saudáveis, com 3-4 meses de idade, 2-3kg de peso, para exploração das glândulas paratireóides, que foram coradas com hematoxilina e eosina (HE) após a remoção. Os seguintes tipos foram classificados de acordo com a relação entre a posição da glândula paratireoidiana inferior e a tireoide: Tipo A, Tipo Próximo, Tipo B e Tipo Distante. Houve 188 casos, 4 em que as glândulas paratireoidianas inferiores estavam localizadas próximas ao lado dorsal da tireoide (2.13%), 8 onde as glândulas paratireoidianas inferiores estavam localizadas superiores ao polo superior da tireoide (4.26%), 20 onde as glândulas paratireoidianas inferiores estavam localizadas paralelo à tireoide (10.64%) e 155 casos em que as glândulas paratireoidianas inferiores estavam localizadas inferiores ao polo inferior da tireoide (82.45%). A identificação da localização e a classificação da vasculatura das glândulas paratireóides em coelhos brancos da Nova Zelândia fornecerão um modelo anatômico para auxiliar em pesquisas futuras.(AU)

Probiotic Lactobacilli Improved Growth Performance and Attenuated Salmonella Typhimurium Infection Via Jak/Stat Signaling in Broilers

This study aimed to investigate the effect of Lactobacillus plantarum DPP8 and Lactobacillus acidophilus C7282 in feed supplementation on growth performance, Salmonella invasion, inflammation, and mediating signaling in broilers infected with Salmonella Typhimurium (S. Typhimurium). A total of 240 broilers at day old were randomly allocated into four groups, orally infected with S. Typhimurium and supplemented with individual or combined Lactobacilli DPP8 and C7282 at doses of 0 (control), 1010 (individual), or 2.0 × 1010 (combination) cfu/kg of diet for 21 d. The results showed that supplementing Lactobacilli improved (p 0.05) feed intake and body weight gain and decreased (p 0.05) S. Typhimurium load in the caecum, harder gland, spleen and bursa of Fabricius. Also, the supplements decreased (p 0.05) interleukin (1/2/4), tumor necrosis factor and interferon in the serum, enhanced (p 0.05) interleukin 10, and downregulated gene expressions of inflammatory mediators including Janus kinase (Jak2/3), signal transducer and activator of transcription protein (STAT3/4/5/6) in the intestinal mucosa. In contrast, diets containing DPP8 exhibited greater effects on the inhibition of the pathogen and inflammatory response than C7282. The obtained data suggest that Lactobacilli C7282 and DPP8 can be used as feed additives to inhibit colonization and translocation of S. Typhimurium and inflammatory responses via downregulating Jak/STAT signaling in broilers.(AU)

Probiotic Lactobacilli Improved Growth Performance and Attenuated Salmonella Typhimurium Infection Via Jak/Stat Signaling in Broilers

This study aimed to investigate the effect of Lactobacillus plantarum DPP8 and Lactobacillus acidophilus C7282 in feed supplementation on growth performance, Salmonella invasion, inflammation, and mediating signaling in broilers infected with Salmonella Typhimurium (S. Typhimurium). A total of 240 broilers at day old were randomly allocated into four groups, orally infected with S. Typhimurium and supplemented with individual or combined Lactobacilli DPP8 and C7282 at doses of 0 (control), 1010 (individual), or 2.0 × 1010 (combination) cfu/kg of diet for 21 d. The results showed that supplementing Lactobacilli improved (p 0.05) feed intake and body weight gain and decreased (p 0.05) S. Typhimurium load in the caecum, harder gland, spleen and bursa of Fabricius. Also, the supplements decreased (p 0.05) interleukin (1/2/4), tumor necrosis factor and interferon in the serum, enhanced (p 0.05) interleukin 10, and downregulated gene expressions of inflammatory mediators including Janus kinase (Jak2/3), signal transducer and activator of transcription protein (STAT3/4/5/6) in the intestinal mucosa. In contrast, diets containing DPP8 exhibited greater effects on the inhibition of the pathogen and inflammatory response than C7282. The obtained data suggest that Lactobacilli C7282 and DPP8 can be used as feed additives to inhibit colonization and translocation of S. Typhimurium and inflammatory responses via downregulating Jak/STAT signaling in broilers.

Dietary Lactic Acid Bacteria Modulate Yolk Components and Cholesterol Metabolism by Hmgr Pathway in Laying Hens

This study aimed to investigate the effect of dietary lactic acid bacteria (LAB) on egg production, yolk components, cholesterol metabolism, and enterohepatic circulation of bile acids in hens. Four treatment diets included a control and LAB added at 3 × 105 (low), 3 × 107 (medium), or 3 × 109 (high) cfu/kg. The treatment LAB contained equal amounts of Lactobacillus acidophilus, Lactobacillus plantarum, and Enterococcus faecium. Results showed that high LAB increased (p 0.05) laying rate, egg mass, and yolk phospholipid, but decreased (p 0.05) yolk triglyceride and phosvitin. Diets with LAB decreased (p 0.05) yolk and serum cholesterol content, and serum bile acid by 9.3 to 39.9%. In liver, high LAB downregulated (p 0.05) mRNA expression of serine/threonine kinase 11 (STK11), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), AMP-activated protein kinase catalytic subunit (PRKAA1, 2), and protein phosphatase catalytic subunits (PPP2CA, PPP2CB and PPP3CA) by 49.5 to 175.4%. In mucosa, high LAB downregulated (p 0.05) PRKAA1 and HMGR by 68.2 and 69.6%, respectively; but upregulated (p 0.05) PPP2CA and PPP2CB by 51.2 and 45%, respectively. Linear decreasing (p0.035) responses to LAB doses were found on cholesterol, phosvitin, bile acid, and hepatic gene expressions, and quadratic (p0.006) effects on yolk cholesterol and hepatic STK11. It is concluded that probiotic LAB can improve yolk components and decrease hepatic cholesterol synthesis by regulating HMGR pathway in hens.(AU)

Dietary Lactic Acid Bacteria Modulate Yolk Components and Cholesterol Metabolism by Hmgr Pathway in Laying Hens

This study aimed to investigate the effect of dietary lactic acid bacteria (LAB) on egg production, yolk components, cholesterol metabolism, and enterohepatic circulation of bile acids in hens. Four treatment diets included a control and LAB added at 3 × 105 (low), 3 × 107 (medium), or 3 × 109 (high) cfu/kg. The treatment LAB contained equal amounts of Lactobacillus acidophilus, Lactobacillus plantarum, and Enterococcus faecium. Results showed that high LAB increased (p 0.05) laying rate, egg mass, and yolk phospholipid, but decreased (p 0.05) yolk triglyceride and phosvitin. Diets with LAB decreased (p 0.05) yolk and serum cholesterol content, and serum bile acid by 9.3 to 39.9%. In liver, high LAB downregulated (p 0.05) mRNA expression of serine/threonine kinase 11 (STK11), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), AMP-activated protein kinase catalytic subunit (PRKAA1, 2), and protein phosphatase catalytic subunits (PPP2CA, PPP2CB and PPP3CA) by 49.5 to 175.4%. In mucosa, high LAB downregulated (p 0.05) PRKAA1 and HMGR by 68.2 and 69.6%, respectively; but upregulated (p 0.05) PPP2CA and PPP2CB by 51.2 and 45%, respectively. Linear decreasing (p0.035) responses to LAB doses were found on cholesterol, phosvitin, bile acid, and hepatic gene expressions, and quadratic (p0.006) effects on yolk cholesterol and hepatic STK11. It is concluded that probiotic LAB can improve yolk components and decrease hepatic cholesterol synthesis by regulating HMGR pathway in hens.

Effect of broccoli residues fermented with probiotics on the growth performance and health status of broilers challenged with Clostridium perfringens

This study aimed at investigating the effects of broccoli residues fermented with probiotics (BF) on the growth performance, immunity, and gut health in broilers challenged with Clostridium perfringens (C. perfringens). A total of 600 broilers (one day old) were randomly allotted into five treatments with six replicates of 20 birds each and were reared until 42 days of age. The treatments included a positive control (PC, fed a basal diet and reared on uncontaminated litter), a negative control (NC, birds reared on litter contaminated with C. perfringens and fed a basal diet), and NC plus BF at 25, 50 or 75 g/kg of diet. The BF contained yeast 3.1 × 10 (7) cfu/g, lactic acid bacteria 9.5 × 10 (6) cfu/g and Bacillus subtilis 3.5 × 10 (6) cfu/g. Birds in the NC group showed lower (p<0.05) feed intake and body weight gain, whereas BF supplementation recovered (p<0.05) the growth performance to the levels of PC group. Dietary BF at 50and 75 g/kg reduced (p<0.05) broiler mortality. Similarly, compared to the NC group, BF increased (p<0.05) immune organ weights and serum immunoglobulins A, G, and M to the levels of PC group. The ileal populations of Escherichia coli and Gram-negative bacteria were decreased (p<0.05) by BF to the levels of PC, and C. perfringens was also decreased (p<0.05) by BF. The serum profiles of mono- and di-amine oxidase were decreased (p<0.05) by BF. BF at 75 g/kg reduced (p<0.05) monoamine oxidase compared with the other BF doses. The results suggest that broccoli residues fermented with probiotics can be a novel biological feed additive to protect the performance and health of broilers against C. perfringens infection.(AU)

Effect of broccoli residues fermented with probiotics on the growth performance and health status of broilers challenged with Clostridium perfringens

This study aimed at investigating the effects of broccoli residues fermented with probiotics (BF) on the growth performance, immunity, and gut health in broilers challenged with Clostridium perfringens (C. perfringens). A total of 600 broilers (one day old) were randomly allotted into five treatments with six replicates of 20 birds each and were reared until 42 days of age. The treatments included a positive control (PC, fed a basal diet and reared on uncontaminated litter), a negative control (NC, birds reared on litter contaminated with C. perfringens and fed a basal diet), and NC plus BF at 25, 50 or 75 g/kg of diet. The BF contained yeast 3.1 × 10 (7) cfu/g, lactic acid bacteria 9.5 × 10 (6) cfu/g and Bacillus subtilis 3.5 × 10 (6) cfu/g. Birds in the NC group showed lower (p<0.05) feed intake and body weight gain, whereas BF supplementation recovered (p<0.05) the growth performance to the levels of PC group. Dietary BF at 50and 75 g/kg reduced (p<0.05) broiler mortality. Similarly, compared to the NC group, BF increased (p<0.05) immune organ weights and serum immunoglobulins A, G, and M to the levels of PC group. The ileal populations of Escherichia coli and Gram-negative bacteria were decreased (p<0.05) by BF to the levels of PC, and C. perfringens was also decreased (p<0.05) by BF. The serum profiles of mono- and di-amine oxidase were decreased (p<0.05) by BF. BF at 75 g/kg reduced (p<0.05) monoamine oxidase compared with the other BF doses. The results suggest that broccoli residues fermented with probiotics can be a novel biological feed additive to protect the performance and health of broilers against C. perfringens infection.

Two novel Mg(II)-based and Zn(II)-based complexes: inhibiting growth of human liver cancer cells

Two new Mg(II)-based and Zn(II)-based coordination polymers, {[Mg3(BTB)(DMA)4](DMA)2}n (1, H3BTB=1,3,5-benzenetrisbenzoic acid, DMA=N,N-dimethylacetamide) and {(H2NMe2)2[Zn3(BTB)2(OH)(Im)](DMF)9(MeOH)7}n (2, Im=imidazole, DMF=N,N-dimethylformamide), have been successfully synthesized and structurally characterized under solvothermal conditions. 1 contains a linear [Mg3(COO)6] cluster that connected by the fully deprotonated BTB3- ligands to give a kgd-type 2D bilayer structure; 2 represents a microporous 3D pillar-layered system based on the binuclear Zn units and pillared Im ligands, which shows a (3,5)-connected hms topological net. In addition, in vitro anticancer activities of compounds 1 and 2 on 4 human liver cancer cells (HB611, HHCC, BEL-7405 and SMMC-7721) were determined.

Two novel Mg(II)-based and Zn(II)-based complexes: inhibiting growth of human liver cancer cells.

Two new Mg(II)-based and Zn(II)-based coordination polymers, {[Mg3(BTB)(DMA)4](DMA)2}n (1, H3BTB=1,3,5-benzenetrisbenzoic acid, DMA=N,N-dimethylacetamide) and {(H2NMe2)2[Zn3(BTB)2(OH)(Im)](DMF)9(MeOH)7}n (2, Im=imidazole, DMF=N,N-dimethylformamide), have been successfully synthesized and structurally characterized under solvothermal conditions. 1 contains a linear [Mg3(COO)6] cluster that connected by the fully deprotonated BTB3- ligands to give a kgd-type 2D bilayer structure; 2 represents a microporous 3D pillar-layered system based on the binuclear Zn units and pillared Im ligands, which shows a (3,5)-connected hms topological net. In addition, in vitro anticancer activities of compounds 1 and 2 on 4 human liver cancer cells (HB611, HHCC, BEL-7405 and SMMC-7721) were determined.

Analysis of genetic diversity and population structure in a tomato (Solanum lycopersicum L.) germplasm collection based on single nucleotide polymorphism markers.

Knowledge of genetic diversity is important to assist breeders in the selection of parental materials and in the design of breeding programs. In this study, we genotyped 348 inbred tomato lines, representing vintage and contemporary fresh-market varieties, by using 52 single nucleotide polymorphisms (SNPs); 45 of these were found to be polymorphic. The average minor allele frequency and unbiased expected heterozygosity were 0.315 and 0.356, respectively. Population structure analysis revealed that contemporary germplasm could be distinctly divided into six subpopulations representing three market classes and breeding programs (pink, green, and red). Vintage germplasm could be separated into at least two subpopulations, and more admixtures were found in vintage lines than in contemporary lines. These findings indicate that contemporary inbred lines are more diversified than vintage inbred lines. AMOVA of vintage and contemporary lines was performed. A significant difference was found (P < 0.01), which explained 17.4% of the total genetic variance. Subsequently, we constructed a core collection using 45 polymorphic SNP markers. The data showed that all alleles were captured by only 2% of lines, indicating that more alleles, as well as rare alleles, could enable more variation to be captured in the core collection. These data allow us to discard redundant inbred tomato lines and to select elite inbred lines, which will accelerate the breeding process.

Endometrial expression of telomerase, progesterone, and estrogen receptors during the implantation window in patients with recurrent implantation failure.

Telomerase plays a critical role in cell proliferation and senescence, but the exact involvement of endometrial telomerase in recurrent implantation failure (RIF) is unknown. We collected endometrial biopsies from RIF patients (N = 30) and fertile women (N = 30). Real-time PCR was performed for detecting changes in telomerase reverse transcriptase (Tert), ER alpha, and PR expression at the transcript level, and the correlation between the variable expressions of these genes was tested using regression analysis. Then, western blot and immunohistochemistry were used to analyze the expression profiles of TERT and ER alpha at the protein level. Compared to the control, Tert expression was substantially increased, whereas ER alpha expression significantly decreased in the endometrium with RIF. No change was observed in PR expression. Tert expression was inversely associated with ER alpha expression. TERT protein expression in RIF patients was also clearly elevated, and was localized to both the endometrial epithelium and stromal cells. However, the signals for ER alpha in the stromal cells were weaker than those in the control. Expression of endometrial telomerase was substantially enhanced as ER alpha decreased in RIF patients during the implantation window.

Alpaca fiber growth is mediated by microRNA let-7b via down-regulation of target gene FGF5.

MicroRNAs are very small endogenous RNA molecules that play a crucial role in an array of biological processes, including regulation of skin morphogenesis. The microRNA let-7b is thought to modulate animal hair growth, by binding target genes that encode growth factors. Fibroblast growth factor 5 (FGF5) has been previously reported to be involved in the initiation of the catagen phase of hair growth. In this study, we combined previous reports with bioinformatic analysis techniques to identify and validate FGF5 and, using lucerifase assay, confirmed targeted binding of let-7b to FGF5. To investigate the interaction between let-7b and FGF5, alpaca skin fibroblasts were transfected with let-7b over-expression vectors, and then mRNA and protein expression levels of FGF5 and the gene encoding its receptor, FGFR1, were evaluated. Levels of FGF5 mRNA and protein were remarkably lower in transfected groups, as compared to controls. In summary, this study confirmed that let-7b acts as a regulator of skin morphogenesis, by directly targeting FGF5 and down-regulating its expression. It provides the evidence of hair growth regulated by miRNAs in animals and may have important applications in wool production.

Dynamic comparison of genetic diversity in a Small Tail Han sheep population using meta-analysis.

The aim of this research was to identify the dynamic diversity of Small Tail Han sheep in its main producing areas between different years, and provide a basis for a breeding and genetic resources conservation strategy. For this purpose, 15 microsatellites were genotyped for Small Tail Han Sheep sampled in 2014 from Heze, China, and a comparative analysis of these data with those from a previous study was undertaken using meta-analysis. The results reveal that inbreeding has caused a reduction in diversity of Small Tail Han Sheep from 2008 to 2014. Overall, our results are helpful in understanding the dynamic change in diversity, as well as providing information for a conservation strategy for this population.

Prenatal diagnosis of Chinese families with phenylketonuria.

The aim of this study is to investigate the ability to prenatally diagnose phenylketonuria (PKU) by using phenylalanine hydroxylase (PAH) gene mutation analysis combined with short tandem repeat (STR) linkage analysis in 118 fetuses from 112 Chinese families. Genomic DNA was extracted from the peripheral blood from members of 112 families and the exons and exon-intron boundaries of the PAH gene were amplified by PCR. PCR products were analyzed by bi-directional Sanger sequencing and multiplex ligation-dependent probe amplification (MLPA). The three variable number of tandem repeat (VNTR) markers PAH-1, PAH-26, PAH-32 were used in the prenatal diagnosis for the PKU families. We identified a spectrum of 63 different mutations, including 61 point mutations and indels, two large exon deletion mutations, and five novel mutations. A substantial proportion of mutant alleles were accounted for by p.R243Q (15.62%), EX6-96AG (9.82%), p.V399V (7.59%), p.Y356X (6.70%), and p.R413P (5.36%). The same mutations were identified in 31 prenatally genotyped fetuses. We identified 58 fetuses that carried only one mutant allele and 29 fetuses that carried no mutations of PAH and were presumed normal. PAH gene mutation analysis combined with STR linkage analysis can provide rapid and accurate prenatal diagnosis for PKU families.

Effects of acupotomylysis on basic fibroblast growth factor and CD34 levels in rabbits with third lumbar vertebral transverse foramen syndrome.

This study observed the local tissue homogenates in rabbits with third lumbar vertebral transverse foramen syndrome and explored the mechanism of acupotomylysis in local tissue revascularization. Thirty Japanese white rabbits were randomly divided into the following 5 groups of 6 rabbits each: normal, model, acupotomy, electroacupuncture (EA), and acupotomy-EA groups. All except the normal group were comprised of animal models of third lumbar vertebral transverse foramen syndrome prepared by embedding sponge in the left third lumbar transverse process. The rabbits in the acupotomy and EA groups underwent bilateral acupotomylysis intervention; those in the acupotomy-EA group underwent acupotomylysis and EA interventions. On the 28th day after modeling, the double-antibody ELISA was used to detect b-FGF and CD34 levels in the serum and homogenates of a muscle tissue sample from the left side of the third lumbar transverse process. The b-FGF levels in local muscle homogenates were significantly higher in the modeled rabbits than in the normal rabbits (P < 0.01), and the CD34 levels in the modeled group were significantly lower than in the normal group (P < 0.01). The b-FGF and CD34 levels in the EA, acutopomy, and acutopomy-EA groups were significantly lower than those in the modeled group (P < 0.01); the CD34 levels were significantly higher in the acupotomy-EA group than in the model group (P < 0.05); and the differences among the EA, acupotomy, and acupotomy-EA groups were not significant (P > 0.05). In conclusion, acupotomylysis regulates the levels of b-FGF and CD34 levels in serum and muscle tissue as well as local tissue revascularization.

Mutation analyses and prenatal diagnosis in families of X-linked severe combined immunodeficiency caused by IL2Rγ gene novel mutation.

We investigated the feasibility of interleukin-2 receptor gamma (IL2Rγ) gene based on gene mutation analysis and pre-natal diagnosis of X-linked severe combined immunodeficiency (X-SCID). Blood samples of patients and their parents of X-SCID (family 1) and X-SCID (family 2) were collected. IL2Rγ gene sequences of the 2 families were analyzed using bi-directional direct sequencing by polymerase chain reaction. DNA sequence changes in the IL2Rγ gene exon region and shear zone were also analyzed. We also sequenced the IL2Rγ gene in 100 healthy individuals. Prenatal genetic diagnoses for a high-risk fetus in family 1 were performed by chorionic villus sampling after determining each family's genotypes. The suspect fe-male in family 1 underwent carrier detection. Two novel mutations of IL2Rγ gene were identified, including c.361-363delGAG (p.E121del) in the patient and his mother in family 1, and c.510-511insGAACT (p.W173X) heterozygous mutation in the proband's mother in family 2. These mutations were absent in the 100 controls. Prenatal diagnosis of early pregnancy in the female fetus of family 1 was performed; the fetus was heterozygous, which was confirmed at postnatal follow-up. The suspect female in family 1 showed no mutation in carrier detection. The novel p.E121del and p.W173X mutations in IL2Rγ may have been the primary causes of disease in 2 families with X-SCID. In couples with an X-SCID reproductive history, prenatal gene mutation analysis of IL2Rγ can effectively prevent the birth of children with X-SCID and carrier detection for suspected females.

Decreased TIM-3 mRNA expression in peripheral blood mononuclear cells from nephropathy patients.

Increasing evidence shows that TIM-1 and TIM-3 in-fluence chronic autoimmune diseases, and their expression levels in immune cells from nephritic patients are still unknown. Real-time transcription-polymerase chain reaction analysis was used to deter-mine expression levels of TIM-1 and TIM-3 mRNA in peripheral blood mononuclear cells (PBMCs) from 36 patients with minimal change glo-merulopathy (MCG), 65 patients with lupus nephritis (LN), 78 patients with IgA nephropathy (IgAN), 55 patients with membranous nephropa-thy (MN), 22 patients with crescentic glomerulonephritis (CGN), 26 patients with anaphylactoid purpura nephritis (APN), and 63 healthy controls. TIM-3 mRNA expression significantly decreased in PBMCs from nephritic patients (LN, P < 0.0001; MCG, P < 0.0001; MN, P = 0.0031; CGN, P = 0.0464; IgAN, P = 0.0002; APN, P = 0.0392) com-pared with healthy controls. In contrast, there was no significant differ-ence in TIM-1 mRNA expression between the patients and the healthy controls. Our results suggest that insufficient expression of TIM-3 mRNA may be involved in the pathogenesis of nephropathy.