Objective To investigate the genetic variation of Eurytrema pancreaticum isolated from goats in Huaihua City, Hunan Province. Methods The partial sequence of mitochondrial cytochrome I (pcox1) and ribosomal 18S rRNA genes were amplified using a PCR assay in E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and the PCR amplification products were sequenced. Then, the gene sequences were subjected to genetic variation and phylogenetic analyses. Results The sequences of the pcox1 and 18S rRNA genes were 430 bp and 1 857 bp in length in 18 E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and there were 14 and 35 variation sites in pcox1 and 18S rRNA gene sequences, with intra-species genetic variations of 0 to 1.4% and 0 to 0.8%, respectively. The sequences of pcox1 and 18S rRNA genes had 99.0% to 99.8% and 99.5% to 99.8% homologies with those from E. pancreaticum Chinese strain recorded in the GenBank database. Consistent phylogenetic analysis results were found based on pcox1 and 18S rRNA genes. The 18 E. pancreaticum isolates from goats in Huaihua City were clustered into a clade with the known E. pancreaticum isolates registered in GenBank, and the clade with these 18 E. pancreaticum isolates was close to the clades with Eurytrema species and far from the clades with other trematodes. Conclusions The E. pancreaticum isolates from goats have a low genetic variation in Huaihua City, Hunan Province. Mitochondrial pcox1 and ribosomal 18S rRNA genes may serve as molecular markers for the studies on the genetic variation in goat-derived E. pancreaticum.

Objective To explore the influence of melatonin receptor agonist Neu-P11 on the expression of IRS-1 and GLUT-4 in insulin-resistant 3T3-L1 adipocytes. Methods Insulin resistant 3T3-L1 adipocytes were induced with high glucose/ high insulin for 24 hours, and then they were divided into 4 treatment groups; melatonin, Neu-P11, melatonin+luzindole and Neu-P11 + luzindole. And non-treated insulin-resistant 3T3-L1 adipocytes were taken as control. Glucose consumption was detected by enzymatic method. IRS-1 and GLUT-4 protein expressions were detected by Western blotting analysis. Results After the insulin-resistant adipocytes were treated with melatonin and Neu-P11, the glucose consumption andthe expressions of IRS-1, GLUT-4 proteinswere significantly increased compared with the non-treated control group (P<0. 05). The expressions of IRS-1, GLUT-4 proteins in melatonin + luzindole (melatonin receptor antagonist) and Neu-P11 + luzindole groups were significantly decreased compared with melatonin alone or Neu-P11 alone groups (P<0. 05), and were similar to those in thenon-treated control group. Conclusion Melatonin receptor agonist Neu-P11 can increase glucose consumption, insulin sensitivity in insulin resistant-adipocytes, which might be associated with the up-regulation of IRS-1 and GLUT-4 protein expression.

Objective To analyze the sequence characteristics of Rhipicephalus microplus Enolase gene, and to predict the secondary and tertiary structure and antigenic epitopes of the Enolase protein. Methods Sixty-two engorged female R. microplus were sampled from a yellow cattle breeding farm in Zhijiang County, Huaihua City, Hunan Province in June 25, 2022. Genomic DNA was isolated from R. microplus, and the Enolase gene was amplified using PCR assay, followed by cloning, sequencing and expression of the amplification product. The sequence characteristics of the Enolase gene were analyzed using the software Clustal X, and the gene sequence was translated into amino acid sequences. The secondary and tertiary structures of the Enolase protein were deduced using the software PRABI, and the physicochemical properties of the Enolase protein were analyzed using the software PRABI. In addition, the B- and T-cell epitopes of the Enolase protein were predicted using the software ABCpred Prediction, Scratch, IEDB and NetCTL. Results The R. microplus Enolase gene sequence was 1 323 bp in size, and the contents of A, T, G and C bases were 24.5%, 22.5%, 27.0% and 26.0%,with 47.0% of A + T content and 53.0% of G + C content. The R. microplus Enolase gene encoded 434 amino acids, and the Enolase protein had a molecular weight of 47.12 kDa. The secondary structure of the Enolase protein contained 186 α-helixes (42.86%), 32 β-turns (7.37%), 144 random coils (33.18%) and 72 extended strands (16.59%). The Enolase protein was most probably present in cytoplasm (76.7%), followed by in mitochondrion (39.1%) and nucleus (21.7%), and the Enolase protein had no signal peptide or transmembrane domain. In addition, the Enolase protein had 14 B-cell dominant epitopes and 8 T-cell dominant epitopes. Conclusions The R. microplus Enolase gene sequence exhibits a GC preference, and its encoding Enolase protein is an acidic and hydrophilic protein, with α-helixes and random coils as its primary structure, and presenting B- and T-cell dominant epitopes, which is a potential target for development of vaccines against R. microplus.

OBJECTIVETo investigate the relationship between four cytokines in serum and prostatic fluid and chronic abacterial prostatitis (CAP).

METHODSThe levels of interleukin-2 (IL-2), IL-4, IL-8, tumor necrosis factor-alpha (TNF-alpha) were detected in 86 patients with CAP [CAP I (n = 60), WBC > or = 10/HP; CAP II (n = 26, WBC < 10/HP)], and 20 healthy males as control by ELISA.

RESULTSIL-2, IL-8 and TNF-alpha in serum and prostatic fluid, and IL-4 in serum in patients with CAP I were markedly higher than those in the control, respectively (P < 0.05 or P < 0.01). No significant difference was observed in IL-4 level in prostatic fluid among CAP I, CAP II and control groups respectively (P > 0.05), and in IL-2, IL4, TNF-alpha in serum and prostatic fluid between CAP II group and the control (P > 0.05). IL-8 was correlated with WBC number in prostatic fluid (r = 0.62, P < 0.05).

CONCLUSIONIL-2, IL-8, TNF-alpha may play roles in the process of pathogenesis of CAP, and they have applicable value in the diagnosis and typing of CAP.

Adult ; Bodily Secretions ; chemistry ; Chronic Disease ; Cytokines ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-8 ; metabolism ; Male ; Middle Aged ; Prostate ; secretion ; Prostatitis ; blood ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the clinical symptoms and potential mutation in FGFR3 gene for a family featuring hereditary dwarfism in order to attain diagnosis and provide prenatal diagnosis.

METHODSFive patients and two unaffected relatives from the family, in addition with 100 healthy controls, were recruited. Genome DNA was extracted. Exons 10 and 13 of the FGFR3 gene were amplified using polymerase chain reaction (PCR). PCR products were sequenced in both directions.

RESULTSAll patients had similar features including short stature, short limbs, lumbar hyperlordosis but normal craniofacial features. A heterozygous mutation G1620T (N540K) was identified in the cDNA from all patients but not in the unaffected relatives and 100 control subjects. A heterozygous G380R mutation was excluded.

CONCLUSIONThe hereditary dwarfism featured by this family has been caused by hypochondroplasia (HCH) due to a N540K mutation in the FGFR3 gene.

Base Sequence ; DNA Mutational Analysis ; Dwarfism ; genetics ; Exons ; Female ; Heterozygote ; Humans ; Male ; Mutation ; Receptor, Fibroblast Growth Factor, Type 3 ; genetics

OBJECTIVETo analyze clinical symptoms and disease-causing mutations of corneodesmosin (CDSN) gene in a Chinese family affected with hypotrichosis simplex of the scalp and to establish a method for prenatal diagnosis.

METHODSFamily survey and clinical examinations were carried out to determine the inheritance pattern. Three patients and 7 unaffected relatives from the family, in addition with 100 unrelated healthy controls were recruited. Genomic DNA from peripheral blood leukocytes was extracted. Five pairs of primers were designed based on the CDSN gene sequence. Exons and flanking regions of the CDSN gene were amplified using polymerase chain reaction (PCR). Potential mutations were analyzed through direct sequencing and comparison by BLAST.

RESULTSThe type of alopecia of the family was diagnosed as hypotrichosis simplex of the scalp with an autosomal dominant inheritance pattern. A nonsense mutation (C717G) in cDNA sequence of the CDSN gene was identified in all three patients of the family, which resulted in a premature stop codon (Y239X). The same mutation was not found among healthy members of the family and 100 healthy controls.

CONCLUSIONA Chinese family was diagnosed with hypotrichosis simplex of the scalp, which was caused by a novel nonsense mutation (Y239X) in the CDSN gene.

Alopecia ; genetics ; China ; Codon, Nonsense ; Female ; Glycoproteins ; genetics ; Humans ; Hypotrichosis ; genetics ; Male ; Middle Aged ; Pedigree ; Scalp

OBJECTIVETo identify the possible mutation at possible sites in different mitochondrial genes that leads to hearing loss in a large Chinese pedigree.

METHODSBlood samples from a Hunan pedigree were obtained with informed consent. Genomic DNA was extracted from peripheral blood leukocytes using kit. The target fragments were amplified and detected by polymerase chain reaction (PCR) and directly sequencing respectively.

RESULTSThe result of direct sequencing revealed the A1555G mutation in 12S rRNA gene was inherited in this pedigree and no one has A7445G mutation or other mutations in its neighborhood region.

CONCLUSIONSequence analysis confirmed that the pedigree carries the A1555G mutation. With some members ever exposure of aminoglycoside antibiotics, mutation of A1555G may play a pivotal role in the pathogenesis of hearing loss in the large pedigree.

Base Sequence ; China ; DNA Mutational Analysis ; DNA, Mitochondrial ; chemistry ; genetics ; Family Health ; Female ; Hearing Loss ; genetics ; Humans ; Male ; Marriage ; Pedigree ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics ; RNA, Transfer, Ser ; genetics

Differences of some points, levels and angles of the healthy and affected sides of patients with peripheral facial paralysis were picked out according to photographs. Through analysis of the index between the healthy and affected side of the patients and the difference between healthy people and patients, it is approved that those special points, levels and angles, which are called as deviation index of eye and mouth, can evaluate peripheral facial paralysis objectively and judge the degree of deviation. Therefore, it provides references for the diagnosis of facial paralysis and its degree judgement.
Adolescent ; Adult ; Aged ; Child ; Eye ; anatomy & histology ; Facial Paralysis ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Mouth ; anatomy & histology

Objective To investigate the level of deltamethrin resistance and mutation sites in the sodium iron channel gene in Rhipicephalus microplus in Huaihua City, Hunan Province, and to examine the correlation between deltamethrin resistance and mutation sites in the sodium iron channel gene in Rh. microplus. Methods Rh. microplus was sampled from multiple yellow cattle farms in Huaihua City, Hunan Province from June to September 2022, and the level of resistance to deltamethrin was determined in ticks using the adult immersion test. The sodium iron channel domain III gene was amplified in deltamethrin-resistant and wild-type Rh. microplus using PCR assay. Following sequencing and sequence alignment, mutation sites were detected in bases. The sodium iron channel domain III gene in Rh. microplus was translated, and the signal peptide, transmembrane domain, and phosphorylation and glycosylation sites were detected in amino acid sequences. The tertiary structures of the sodium iron channel domain III protein of deltamethrin-resistant and wild-type Rh. microplus were deduced and compared, and the association be tween mutation sites in bases and resistance to deltamethrin was examined in Rh. microplus according the level of deltamethrin resistance, sequence alignment and protein tertiary structure. Results The median (LC₅₀) and 95% lethal concentrations (LC₉₅) of deltamethrin were 121.39 mg/L and 952.61 mg/L against Rh. microplus, with a resistance factor of 9.24 and level II resistance. The sequence of the sodium ion channel domain III gene was 1 010 bp in size, and mutation sites were detected in two neighboring bases in the sequence of the sodium ion channel domain III gene in deltamethrin-resistant Rh. microplus. Although no signal peptides were found in the sodium iron channel domain III protein of deltamethrin-resistant or wild-type Rh. microplus, 6 trans-membrane domains, 42 phosphorylation sites and 8 glycosylation sites were identified, with a significant difference in the tertiary structure of the sodium iron channel domain III protein between deltamethrin-resistant and wild-type Rh. microplus. Conclusions Level II resistance to deltamethrin is detected in Rh. microplus in Huaihua City, Hunan Province, and two mutation sites that correlate with the emergence of deltamethrin resistance are identified in the sequence of the sodium iron channel domain III gene in deltamethrin-resistant Rh. microplus.