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.

Objective To assess polymerase chain reaction(PCR)combined with restriction fragment length polymorphism(RFLP)and gene sequencing technologies in the detection and typing of HPV DNA.Methods Tissue specimens were collected from skin diseases and venereal disease in perianal or genitals.PCR was performed with HPV DNA general primers(MY09/11)in tissue samples. Positive fragments of HPV DNA were purified and digested by restriction enzymes.The digested fragments were typed by po]yacrylamide gel electrophoresis(PAGE).The Resultswere verified by direct sequencing.Results In 50 clinical samples there were 35 HPV DNA positive,including 26 from patients with condyloma acuminatum,8 from patients with bowenoid papulosis,and 1 from patients with squamous cell carcinoma.In HPV DNA positive samples,19 were HPV6,3 were HPV11,8 were HPV16,4 were HPV6 and HPV 11,and I was HPV62.Sequencing Resultswere in accordance with the PCR-RFLP Results .Conclusion PCRRFLP method is effective in the detection and typing of HPV DNA.

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.

BACKGROUND:Recently, there are many documents al over the world reporting hypotoxicity infection after fracture internal fixation surgery, but reports are different on whether it is necessary for chronic hypotoxicity infection internal fixation removal post surgery. There are no fixed judgment criteria of curative effects, which leads to inexact conclusion of treatment method. ＜br> OBJECTIVE:To observe the curative effects of taking rifampicin and ciprofloxacin with transfer of skin flap in the treatment of chronic hypotoxicity infection with sinus formation after fracture surgery on tibial plateau and ankle. METHODS:A total of 56 cases of chronic hypotoxicity infection after fracture surgery of tibial plateau and ankle were col ected from September 2005 to December 2012. 30 cases in the therapy group were treated with ＜br> levofloxacin and rifampicin with transfer of skin flap. 26 cases in the control group were treated with conventional intravenous antibiotics with local debridement to remove internal fixation. In both groups, the course of disease was 3 to 6 months. The erythrocyte sedimentation rate, kidney function and radiographic indices were reviewed every 1 to 3 months, and curative effects were evaluated. ＜br> RESULTS AND CONCLUSION:During the fol ow-up visit of 6-24 months, no recurrence happened to the cured and improved patients. In the therapy group, 24 cases were cured, 4 cases were improved, and 2 cases were invalid, with a total effective rate of 93%. In the control group, 11 cases were cured, 6 cases were improved and 9 cases were invalid with the total effective rate of 65%.χ2 test showed that therapeutic effects were significantly better in the therapy group than in the control group (P＜0.05). These data indicated that rifampicin and ciprofloxacin with transfer of skin flap for chronic hypotoxicity infection with sinus formation after fracture surgery of tibial plateau and ankle showed good curative effects.

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

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 trace changes in the transcript level of the Treponema pallidum(Tp)protein Tp0751 in skin lesions of a rabbit model of early syphilis. Methods Three New Zealand white rabbits were intracutaneously injected with 0.1 ml of Tp (Nichols Seattle strains)suspensions (107 treponemes/ml)at 10 sites on the shaved back to establish a model of early syphilis. All the rabbits received a single injection with the total amount of treponemes being 107. Then, skin changes at injection sites were observed, and the size of skin rashes was recorded on a daily basis. Skin specimens sized 0.4 cm × 0.4 cm were excised from an injection site and a non-injection site(negative control)separately every 3 days for the detection of Tp0751 and Tp0574 mRNAs. The whole experiment lasted 30 days, and a total of 11 skin biopsies were carried out. Fluorescence-based quantitative PCR was performed to measure the mRNA expressions of Tp0751 and Tp0574 continuously and dynamically during the development of chancre. Results After intracutaneous injection of Tp suspensions, red papules occurred on the back of rabbits on day 6, and reached maximum size on day 19 with the formation of ulcer and chancre. On day 25, disseminated secondary syphilides gradually appeared all over the body surface of the rabbits. The mRNA expression levels of Tp0574 and Tp0751 increased at the early stage, peaked onday 15 (compared with the other time points, all P < 0.05), thereafter rapidly declined, but rose slightly on day 27. The standardized expression level of Tp0751 mRNA increased gradually after day 15, and peaked on day 24 (compared with the other time points, all P < 0.05). Conclusion The transcript level of Tp0751 was high in rabbits at the late stage of Tp clearance when generalized disseminated secondary syphilides had not appeared, suggesting that Tp0751 may be involved in the systemic spread of Tp.

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 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.