With the improvement of living standards,at least a quarter of the global population has non-alcoholic fatty liver disease(NAFLD),which is considered to be an important cause of the increased incidence of hepatocellular carcinoma(HCC)in recent years.Finding effective means for disease prevention and/or treatment largely relies on deep understanding of the mechanisms of NAFLD to HCC,which needs to con-struct the stable experimental models to simulate the entire process of disease progression in human NAFLD-HCC.This paper summarizes the animal models which are currently used to study NAFLD-HCC and their ad-vantages and disadvantages,in order to provide a basis for the selection of animal models and accelerate the transition from basic study to clinical study.

OBJECTIVE: To explore the genetic basis for 4 patients with globozoospermia. METHODS: Semen and blood samples were collected from the patients for the determination of sperm concentration, viability, survival rate, morphology and acrosome antigen CD46. Meanwhile, DNA was extracted for whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing. RESULTS: All of the four patients were found to harbor variants of the DPY19L2 gene. Patients 1 ~ 3 had homozygous deletions of the DPY19L2 gene. Sanger sequencing confirmed that the DPY19L2 gene in patient 3 was disrupted at a recombination breakpoint area BP2, resulting in nonallelic homologous recombination and complete deletion of the DPY19L2 gene. Patients 2 and 3 respectively harbored novel homozygous deletions of exons 2 ~ 22 and exons 14 ~ 15. Patient 4 harbored heterozygous deletion of the DPY19L2 gene, in addition with a rare homozygous deletion of the 3' UTR region. CONCLUSION DPY19L2 gene variants probably underlay the globozoospermia in the four patients, which has fit an autosomal recessive pattern of inheritance and the characteristics of genomic diseases.
Male ; Humans ; Teratozoospermia/genetics* ; Homozygote ; Semen ; Sequence Deletion ; 3' Untranslated Regions ; Membrane Proteins

Objective To investigate whether the next-generation probiotics Akkermansia muciniphila can prevent non-alcoholic steatohepatitis (NASH)-related hepatocellular carcinoma (HCC). Methods We constructed a NASH-HCC model called STAM. STAM mice received oral saline or A. muciniphila starting at 4 weeks of age. Liver tissues were evaluated by HE staining and oil red O staining for NASH activity, and intrahepatic expression levels of inflammatory cytokines and ileal tight junction proteins were measured by RT-PCR. Results At 8 weeks of age, the steatosis, ballooning degeneration and NAS scores, TNF-α, MCP-1, IL-1β, and IL-6 mRNA expression were significantly decreased in the STAM+A. muciniphila group (both P < 0.05) compared with those in the STAM+saline group (all P < 0.05). At 20 weeks of age, the number of liver surface tumors formed, tumor size and IL-6 level were decreased in the STAM + A. muciniphila group (all P < 0.05). A. muciniphila restored the thickness of the colon mucosal layer and the number of goblet cells in STAM mice as well as increased the mRNA expression of the tight junction proteins ZO-1, Claudin-3, and Occludin in ileal epithelial cells. Conclusion Akkermansia muciniphila can inhibit the progression of NASH to HCC by improving the intestinal barrier function and may serve as a candidate drug to prevent NASH-HCC.

Objective:To establish a new molecular typing method for Treponema pallidum (TP) based on TP0136 protein sequence heterogeneity. Methods:The amino acid sequences of TP0136 open reading frame (ORF) of 9 strains of Treponema pallidum ssp. Pallidum (TPA) , 3 strains of Treponema pallidum ssp. Pertenue (TPE) , 1 unclassified simian strain of Treponema Fribourg-Blanc (FB) and 1 strain of Treponema pallidum ssp. Endemicum (TEN) were searched from Genbank, and multiple sequence comparisons were performed to obtain the molecular typing results of TP0136 protein. The TP0136 protein-based molecular typing method was used to classify 23 TPA clinical isolates, which were collected from Dermatology Hospital of Southern Medical University from January 2015 to December 2018, and the typing results were compared with those by the traditional typing method based on the tp0548/Arp/Tpr genes. Results:TP0136 protein was highly heterogeneous in different TP strains. According to the amino acid sequence of TP0136, TPE, FB and TEN strains were divided into 4 subtypes of Ⅰ- Ⅳ, TPA strains were divided into 6 subtypes of Ⅴ-Ⅹ, and TPA clinical strains were classified into 4 subtypes of Ⅶ, Ⅸ, Ⅹ, Ⅺ. Through the traditional typing method described above, 23 TPA clinical strains could be divided into 5 types (13D/d, 14D/f, 14D/g, 15D/f, 16A/e) . By using the TP0136 protein-based typing method combined with traditional typing method, the above clinical strains could be further subdivided into 10 types, and the 14D/f type could be further divided into 3 subtypes by using the TP0136 protein-based typing method.Conclusion:The TP0136 protein-based molecular typing method can be used to distinguish TP species, which is helpful for further improvement of traditional TPA molecular typing.

Objective To investigate the clinical value of platelet parameters in patients with acute myocardial infarction(AMI) in plateau.Methods A total of 72 patients diagnosed as acute myocardial infarction in our department from January 2016 to June 2017 were enrolled into this study.Clinical data and outcomes were analyzed.Platelet parameters were measured within 24 h after AMI occurrence.The relationship between platelet distribution width (PDW),mean platelet volume (MPV),and the severity of disease,infarct size as well as short-term prognosis were further investigated.Results Compared with control group,PDW and MPV were positively correlated with the severity of disease (PPDW=0.039,PMPV=0.038) and infarct size (rPDW=0.305,P=0.009;rMPV=0.263,P=0.025).The AUC of PDW was 0.827,optimal operating point (OOP) was 16.3％,the AUC of MPV was 0.813,OOP was 13.1 fl,the AUC of GRACE was 0.865,OOP was 145.Conclusions PDW and MPV could be regarded as laboratory index to evaluate the severity of disease,infarct size,pathological changes of coronary artery and short-term prognosis of acute myocardial infarction in plateau.

Objective To investigate the efficacy of double cannula with an early oral diet as nutritional support treatment on non-high output enterocutaneous fistulas. Methods Clinical data from patients with non-high output enterocutaneous fistula, who treated with double cannula with an early oral diet (n = 39) and double cannula with enteral nutrition therapy (n = 43), were retrospectively analyzed. The fistulas status and its closure time were recorded. In order to investigate the effect of oral diet on fistula, the inflammation index and nutritional status indicators before and after therapy were recorded. Results Totally 33 cases of the 39 patients with non-high output fistula were cured after therapy on the average (16.4土7.2) days. The distal non-high output fistula patients had better results(93.1％ closed, 15.8 days)than that of the proximal fistula patients(60.0％, 19.8 days). Laboratory inflammation index(TNF-alpha, IL-6, plasma endotoxin and C-reactive protein) were improved significantly (P＜ 0.05) after therapy. The nutrition status of these patients, such as body weight, hemoglobin, serum total protein, albumin, transferrin, and prealbumin increased at the end of fistula compared to that at the beginning of treatment (P＜ 0.05). Prealbumin and retinol binding protein were significantly higher than that in the control group (P＜0.05). Conclusion With double cannula treatment, early normal eating for enteral nutrition is safe and effective, especially in distal enterocutaneous fistulas. Normal oral diet can achieve the results of fistula closure, reduction of the inflammatory reaction, improvement of the nutrition status, avoiding the infliction of the operation and lowering the cost.

Objective To isolate and culture a clinical strain (GDI) of Treponema pallidum (Tp) in Guangdong province,and to investigate the difference in nonsynonymous single nucleotide polymorphisms (nsSNPs) between the GD1 strain and Tp Nichols strain.Methods The GD1 strain was isolated from the hard chancre in a patient with primary syphilis in Guangdong province,and continuously subcultured in the testes of New Zealand white rabbit.The serial subcultivation of GD1 was multi-verified by dark-field microscopy,polymerase chain reaction (PCR) for TP0548 gene,DNA sequencing and genotyping.Meglumine diatrizoate density gradient centrifugation was performed to isolate rabbit tissues and concentrate GD1,and DNA sequencing was used to verify the nsSNPs in the TP0443 and TP0584 genes.Results The GD1 strain was successfully isolated from the lesions of the patient with syphilis,and classified as a subtype f of TP0548.Compared with the American Tp (Nichols strain),there were nsSNP mutations in the GD1 strain.One mutation was located in the TP0443 gene,leading to the the substitution of threonine by alanine at amino acid position 120,and another one was located in the TP0584 gene,which caused a change from alanine to threonine at amino acid position 314.Conclusion The GD1 strain was successfully isolated firstly from the lesions in a patient with syphilis in Guangdong province,and nsSNP mutations were confirmed in the GD 1 strain on the etiology.

Objective To investigate the expression of TP 0155 mRNA in rabbits with early infection of Treponema pallidum ( T.pallidum ). Methods Three New Zealand white rabbits were subcutaneously injected with T.pallidum Nichols Seattle strains.Each rabbit was inoculated at ten sites with 106 T.pallidum/site.Skin lesions at the primary stage of syphilis were observed at different time points. Biopsy from one of the lesions was obtained from each rabbit every three days for detection of TP 0155 mRNA and house keeping gene TP 0574 mRNA.TP0155 plasmid standard was constructed by molecular cloning technique , and the quantitative PCR was used to continuously detect the expression of TP 0155 mRNA and TP0574 mRNA from lesion at different time points.Kruskal Wallis test and Bonferroni method were used to analyze the data.Results On d6, red papules appeared on the dorsal skins of rabbits ,there were ulcers in the center of the lesions on d19,presenting typical appearance of syphilis chancre.On d24 the scab of ulcer became smaller; on d25 the rabbits showed disseminated secondary syphilis , which became smaller and disappeared on d30.The copy numbers of TP0155 plasmid standards were 7.48 ×109 copies/μL.There were significant differences in expression of both TP 0155 mRNA and TP0574 mRNA at different time points (χ2 =32.756 and 52.344,both P<0.01).The expression levels of TP0155 mRNA and TP0574 mRNA increased in the early stage, and both reached the peak at d15 (both P<0.05), and then rapidly declined. There were significant differences in normalized TP 0155 mRNA ( TP0155 ×1000/TP0574 mRNA ) at different time points(χ2 =19.758,P<0.05),which reached the peak on d24 and d30,respectively (all P<0.05).Conclusion The level of TP0155 mRNA increases with the disappearance of chancre and secondary syphilis lesions , suggesting that TP0155 might be involved in immune escape of T.pallidum.

Objective To investigate the application of venereal disease research laboratory(VDRL) test and toluidine red unheated serum test(TRUST) in syphilis laboratory diagnosis.Methods Serum,plasma and cerebrospinal fluid (CSF) in syphilis patients and healthy controls were measured by VDRL and TRUST.Results The VDRL detection results in serum and CSF sepcimens were generally higher than the TRUST detection results by 1-2 titers,while in plasma specimen,the VDRL detection results were generally lower than the TRUST detection results by 1-2 titers than TRUST when using plasma specimen.In addition,the VDRL detection in normal control serum and plasma specimens all appeared different degrees of false positive,but in the detection of normal control CSF,the results of TRUST and VDRL were consistent.Conclusion TRUST is more suitable for serum and plasma specimens,and CSF is suitable for the CSF specimen,but not suitable for serum and plasma detection.

OBJECTIVETo investigate the effect of anti-miR-145 on human airway smooth muscle cell (HASMC) proliferation and osteopontin systhesis in vitro and explore the mechanisms.

METHODSHASMCs were treated with 10-100 nmol/L anti-miR-145, and the cell proliferation and apoptosis were investigated using a CCK-8 assay and flow cytometry, respectively. The changes in osteopontin synthesis after the treatment was quantified with Western blotting.

RESULTSTreatment with 10 and 50 nmol/L anti-miR-145 significantly promoted the proliferation and osteopontin synthesis in HASMCs (P<0.05 or <0.01), and 50 nmol/L anti-miR-145 obviously inhibited the cell apoptosis (P<0.01).

CONCLUSIONAnti-miR-145 promotes HASMC proliferation and osteopontin synthesis and inhibits HASMC apoptosis in vitro, indicating the important role of anti-miR-145 in the pathogenesis of airway remodeling.

Airway Remodeling ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Humans ; MicroRNAs ; antagonists & inhibitors ; Myocytes, Smooth Muscle ; drug effects ; Osteopontin ; biosynthesis ; Respiratory System ; cytology