With reverse transcriptase PCR, the transcripton of copper homeostasis relative gene Afe0329 in Acidithiobacillus ferrooxians standard strain ATCC23270 was investigated. The further analysis of genes in this transcripton was analyzed employed by Vector NTI, Blast, TMHMM Server, PSORTb software and so on. From the DNA of different strains, the transcripton of Afe0329 was amplified using special primer pairs to identify the universality of it in the genome of A.ferrooxidans strains. The results showed that gene Afe0330 and Afe0331 were cotranscribed with Afe0329, and they were in a single transcripton. Gene Afe0329 was supported to express a P1b3-type ATPase which is a heavy metal ion pumping transmembrane protein, protein AFE0330 which expressed by gene Afe0330 was a cytoplasmic protein, no significant ho- mologous sequences of Afe0330 or Afe0331 had been obtained by Blast analysis. And the transcripton of Afe0329 was universal in genome of A. ferrooxidans strains.

BACKGROUND: There is growing evidence that 5-fluorouracil (5-FU) combined with therapeutic trauma can effectively induce skin repigmentation in vitiligo patients who are unresponsive to conventional treatments. Previous studies have mainly focused on identifying the antimitotic activity of 5-FU for the treatment of skin cancer, but few studies have investigated its extra-genotoxic actions favoring melanocyte recruitment. METHODS: We utilized the full thickness excisional skin wound model in Dct-LacZ transgenic mice to dynamically assess the migration of melanocytes in the margins of wounds treated with or without 5-FU. The in-situ expression of CXCL12 was examined in the wound beds using immunofluorescence staining. Quantitative real-time polymerase chain reaction and Western blotting analyses were performed to detect the expression levels of CXCL12 mRNA and protein in primary mouse dermal fibroblasts treated with or without 5-FU. Transwell assays and fluorescein isothiocyanate (FITC)-phalloidin staining were used to observe cell migration and filamentous actin (F-actin) changes of melan-a murine melanocytes. RESULTS: Whole mount and cryosection X-gal staining showed that the cell numbers of LacZ-positive melanocytes were much higher in the margins of dorsal and tail skin wounds treated with 5-FU compared with the controls. Meanwhile, CXCL12 immunostaining was significantly increased in the dermal compartment of wounds treated with 5-FU (control vs. 5-FU, 22.47 ± 8.85 vs. 44.69 ± 5.97, P < 0.05). Moreover, 5-FU significantly upregulated the expression levels of CXCL12 mRNA (control vs. 5-FU, 1.00 ± 0.08 vs. 1.54 ± 0.06, P < 0.05) and protein (control vs. 5-FU, 1.00 ± 0.06 vs. 2.93 ± 0.10, P < 0.05) in cultured fibroblasts. Inhibition of the CXCL12/CXCR4 axis suppressed melanocyte migration in vitro using a CXCL12 small interfering RNA (siRNA) or a CXCR4 antagonist (AMD3100). CONCLUSION 5-FU possesses a pro-pigmentary activity through activation of the CXCL12/CXCR4 axis to drive the chemotactic migration of melanocytes.
Animals ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12/genetics* ; Fibroblasts ; Fluorouracil/therapeutic use* ; Humans ; Mice ; RNA, Messenger ; Receptors, CXCR4

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

OBJECTIVETo explore the treatment of hypospadias with hydrocele.

METHODSNine patients with hypospadias complicated by hydrocele underwent urethroplasty with tunica vaginalis flap.

RESULTSSatisfactory results were achieved in 7 cases, all with a normal anatomic position of urethral meatus, perfect urination and no infection. Local infection and meatal stricture occurred in 1 case, which was restored to normal by urethral sounding. Scrotal urinary fistula developed in another.

CONCLUSIONWith minimum surgical trauma, conveniently available material and fast tissue repair, urethroplasty with tunica vaginalis flap is a desirable method for the treatment of hypospadias with hydrocele.

Child ; Child, Preschool ; Follow-Up Studies ; Humans ; Hypospadias ; complications ; surgery ; Male ; Surgical Flaps ; Testicular Hydrocele ; complications ; surgery ; Testis ; surgery ; Treatment Outcome ; Urologic Surgical Procedures, Male ; methods

Objective To investigate the effect of silment information regulator factor related enzymes 1 (SIRT1) on the apoptosis of retinal ganglion cells (RGCs) in rats with diabetic retinopathy and its downstream molecular mechanisms.Methods Together 60 healthy male Sprague-Dawley rats were collected and randomly divided into normal group,diabetic group,SIRTI activator-resveratrol treatment group (treatment group),and diabetic rat model was induced by intraperitoneal injection of streptozotocin at 60 mg · kg-1 in the latter two group rats,while the normal group was injected with sodium citrate buffer at 60 mg · kg-1.Then,after 72 h,rats with blood glucose ＞ 16.7 mmol · L-1 were designated as diabetic rats by blood glucose test.Then each rat in the treatment group was treated with SIRT1 activator-resveratrol at 20 g · kg-1 once a day at the 2nd day after the success of the model,and the normal group and diabetic group were given methylene chloride.Finally,after immunohistochemical staining for retina,TUNEL assay was used to evaluate the apoptosis of RGCs,while the expression of SIRTI,p38 MAPK and Caspase-3 protein was detected by Western blot.Results The apoptotic index of RGCs in the normal group,diabetic group and treatment group was (0.848+0.131)％,(19.038 + 1.327)％,(10.461 + 1.089)％ respectively at 8 weeks,and the difference among the three groups was statistically significant (F =670.497,P =0.000),while the differences between each two groups were also statistically significant (all P =0.000).Furthermore,when compared with the normal group (0.132 ± 0.043),the expression of SIRT1 protein in the diabetic group (0.060 ± 0.028) and the treatment group (0.073 ± 0.026) was significantly decreased,and the overall difference among the three groups was statistically significant (F =1 310.663,P =0.000),while the differences between each two groups were also statistically significant (all P =0.000).The expression levels of p38 MAPK and Caspase-3 were increased in diabetic group (1.121 ± 0.082,0.266 ± 0.005) and treatment group (0.574 ± 0.012,0.190 ±0.060) respectively,and the overall difference and pairwise comparison in the three groups approached statistically significance (all P =0.000,0.000).Conelusion Up-regulation of SIRT1,can inhibit the apoptosis of RGCs,and protect RGCs against apoptosis in rat model of diabetic retinopathy,which may be correlated with the downregulation of p38 MAPK signal pathway.

OBJECTIVETo assess the clinic efficacy and safety in the treatment of chronic abacterial prostatitis/chronic pelvic pain syndrome (CABP/CPPS) after intraprostatic injection of Chuanshentong.

METHODSFive milliliter of solution blending Chuanshentong and lidocaine was transperineally injected into one lobe of prostate, once a day for 6 days, for a total of 98 cases of patients who had been diagnosed as chronic abacterial prostatitis/chronic pelvic pain syndrome. The efficacy was evaluated by the NIH Chronic Prostatitis Symptom Index (CPSI) after a 12 week follow-up.

RESULTSAll cases had completed the follow-up. Fifty-three cases (54.08%) were completely cured, accompanied by remarkable effective in 17 cases (17.35%) and improved in 23 cases (23.47%). The total remarkable efficacy was 71.43% and improved 94. 90%. Scores of CPSI decreased significantly compared with pre-treatment (95% confidence interval 12.85 approximately 17.91).

CONCLUSIONIt is suggested that transperineally intraprostatic injection of Chuanshentong may be a useful method for the treatment of CABP/CPPS.

Adult ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Injections, Intralesional ; Male ; Middle Aged ; Pelvic Pain ; drug therapy ; Phytotherapy ; Prostatitis ; drug therapy

Patients with late-stage cancer commonly have distant lymph node metastasis; however, poor health often contraindicates surgical treatment. Although the quality of life and overall survival for these patients are low, there is neither a consensus nor a guide for treatment. Ablation technique and surrounding tissue damage are two possible reasons for limited study of radiofrequency ablation in patients with superficial distant lymph node metastasis. Here, we report two patients treated successfully with ultrasound-guided radiofrequency ablation for superficial distant lymph node metastasis. In these patients, deionized water was injected to the surrounding tissues of the lymph node to decrease heat injury. Results from these patients suggest that radiofrequency ablation may play an important role in the treatment of patients with distant lymph node metastasis.
Catheter Ablation ; methods ; Esophageal Neoplasms ; diagnostic imaging ; pathology ; surgery ; Female ; Humans ; Lymph Nodes ; diagnostic imaging ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnostic imaging ; pathology ; surgery ; Neoplasm Staging ; Ultrasonography, Interventional ; methods

OBJECTIVETo develop a primer-extension in combination with denaturing high-performance liquid chromatography (PE-DHPLC)-based assay for prenatal diagnosis of the five most common beta-thalassemia mutations in Chinese.

METHODSThe human beta-globin gene fragment was amplified by PCR, followed by a multiple PE reaction specific for each five mutations. Then the PE product mixtures were separated for genotyping of beta-globin gene mutations using fully-denaturing DHPLC analysis.

RESULTSIn a blind study, prenatal diagnosis was performed on thirty-six at-risk families for beta-thalassemia major. Reverse dot blot (RDB) analysis was used to validate each result, showing an accuracy rate of 100% for PE-DHPLC in a total of 108 samples tested. Overall, by PE-DHPLC analysis, the authors could identify the genotypes involving the five mutations and normal alleles corresponding to 94.4% (102/108) and actually make final decision for prenatal diagnosis covering 97.2% (35/36).

CONCLUSIONThe PE-DHPLC protocol can be a simple, rapid, and highly accurate assay in the prenatal detection of common beta-thalassemia mutations.

Base Sequence ; Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; methods ; DNA Primers ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genotype ; Globins ; genetics ; Humans ; Molecular Sequence Data ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; beta-Thalassemia ; diagnosis ; genetics

Objective:To explore whether executive function (EF) deficits are specific to attention-deficit/hyperactivity disorder (ADHD) or whether such deficits are also associated with tics disorder (TD).Methods:A total of 112 ADHD comorbid TD (ADHD +TD) children,112 pure ADHD children and 112 normal controls were selected.The diagnosis was according to the criteria of Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition (DSM-Ⅳ).The groups were matched by sex,age (less than 6 months) and IQ.The Rey-Osterrieth complex Figure Test,the Trail Making Test,and Stroop Color Word Test were administered to assess working memory,shifting and inhibitory function respectively.The results of the comparison between groups used multivariate analysis of variance,and the statistically significant indicators were conducted with apost-test comparison.Results:Both pure ADHD group and ADHD + TD group performed worse (P ＜ 0.05) in the aspects of the delay recalling structure score,the immediate memory detail score,delay recalling detail score of Rey complex figure test,time of numberletter part and shifting time of trail making test,the time of Stroops2,4,the errors of Stroop 4 and word interference than normal controls.In the aspects of the score of the structure and detail forgetting,the pure ADHD group performed worse (P ＜0.05) than normal controls,but the ADHD + TD group didn't.The differences were significant (P ＜ 0.05).The other differences all weren't significant (P ＞ 0.05).Conclusion:It suggests that children with ADHD have executive function deficit,particular in the tests assessing working memory,shifting and inhibitory function,whether or not comorbid tics disorder.Comorbid tics disorders may not be the factor of aggravation.

ObjectiveTo analyze the expression of Lactate dehydrogenase A（LDHA） in both renal cell carcinoma （RCC） tissue and RCC cell lines， and to investigate the impact of LDHA expression on the progression of RCC. MethodsFrom June 2018 to June 2022， totally 52 cases of RCC tissue samples and 49 cases of para-cancerous tissue samples were collected through surgical procedures from our hospital. LDHA expression was detected using immunohistochemistry （IHC）. The expression levels of LDHA in vitro were also detected in the normal human proximal tubule epithelial cell line HK-2 and renal cell carcinoma cell lines A498， Caki-2， ACHN， and 786-O by using qRT-PCR and Western blot. A recombinant plasmid carrying LDHA-shRNA was constructed and then transfected into 786-O cells to down-regulate the expression of LDHA. Tumor proliferative capacity was monitored using CCK-8 assay， clonal formation assay and EdU assessments. Additionally， cell glycolytic activity was assessed through glucose uptake assay， lactate secretion assay， and ECAR analysis. ResultsIHC analysis revealed significantly higher expression of LDHA in RCC tissue compared to adjacent tissues（P<0.05）. Furthermore， RCC tissues with higher TNM stage exhibited greater expression of LDHA than those with lower TNM stage （P<0.05）. The results of qRT-PCR and Western blot demonstrated that the expression of LDHA in each RCC cell line was significantly higher than that in HK-2（P<0.05）. After blocking the expression of LDHA in 786-O， there was a significant down-regulation of cell proliferation and glycolysis capacity （P<0.05）. ConclusionsThe expression of LDHA in RCC tissue and RCC cell lines is significantly overexpressed compared with normal one， particularly in those with higher TNM stage. Knockdown of the expression of LDHA significantly suppresses cell proliferation and aerobic glycolysis capacity in 786-O.