ObjectiveTo determine the genotypes of human papillomavirus(HPV) in patients with giant condyloma acuminatum.MethodsSixty-seven outpatients with giant condyloma acuminatum collected from January 2007 to January 2010 were included in this study.Lesional specimens were obtained from these patients.The genotypes of HPV were determined by flow-through hybridization and gene chip assay.ResultsOf the 67 cases of giant condyloma acuminatum,63 (94.02％) were positive for HPV DNA.Among the HPV DNA-positive specimens,84 (60.87％) harbored low risk types of HPV,54 (39.13％) high risk types of HPV.Type 6 and 11were the predominant low risk HPV types,while type 16 and 18 were the major high risk HPV types.

We constructed plasmid pMTac to overexpress 3-ketosteroid-Δ1-dehydrogenase (KSDD) in Mycobacterium neoaurum JC-12 for improving androst-1,4-diene-3,17-dione (ADD) production. To construct pMTac, pACE promoter on pMF41 was replaced by tac promoter, and then four recombinants were constructed, which were M. neoaurum JC-12/pMF41-gfp, M. neoaurum JC-12/pMTac-gfp, M. neoaurum JC-12/pMF41-ksdd and M. neoaurum JC-12/pMTac-ksdd. Fluorescence detection results show that much more green fluorescent protein (GFP) was expressed in M. neoaurum JC-12/pMTac-ksdd than M. neoaurum JC-12/pMF41-ksdd. The activity of KSDD was 2.41 U/mg in M. neoaurum JC-12/pMTac-ksdd, 6.53-fold as that of M. neoaurum JC-12 and 4.36-fold as that of M. neoaurum JC-12/pMF41-ksdd. In shake flask fermentation, ADD production of M. neoaurum JC-12/pMTac-ksdd was 5.94 g/L, increased about 22.2% compared to the original strain M. neoaurum JC-12 and 12.7% to M. neoaurum JC-12/pMF41-ksdd. AD (4-androstene-3,17-dione) production of JC-12/pMTac-ksdd was 0.17 g/L, decreased 81.5% compared to M. neoaurum JC-12 and 71.2% to M neoaurum JC-12/pMF41-ksdd. In the 5 L fermenter, 20 g/L phytosterols was used as substrate, ADD production of M. neoaurum JC-12/pMTac-ksdd was improved to 10.28 g/L. pMTac is favorable for expressing KSDD in M. neoaurum JC-12, and overexpression of KSDD has beneficial effect on ADD producing, and it is the highest level ever reported using fermentation method in M. neoaurum.
Androstadienes ; metabolism ; Fermentation ; Industrial Microbiology ; Mycobacterium ; Oxidoreductases ; genetics ; metabolism ; Phytosterols ; metabolism ; Plasmids

Objective To report a Chinese pedigree with benign familial chronic pemphigus (BFCP),and to screen mutations of ATP2C 1 gene in this family.Methods A 39-year-old male patient with BFCP andhis family members underwent a clinical investigation.Blood samples were collected from all the members in this family and from 50 unrelated healthy controls.Genomic DNA was extracted from the blood samples,and PCR was performed to amplify all the 28 exons and flanking sequences of the ATP2C1 gene followed by DNA direct sequencing.The resulted DNA sequences were compared with the reported sequences of APT2C1 gene in Genbank (Number:NM_014382.2 and NC_000003.9).Results There were 24 family members in the four-generation pedigree,with 8 members affected by BFCP.A single-nucleotide substitution,c(1696C→T),in exon 17 of the ATP2C1 gene was identified in all of the members with BFCP,but not in unaffected third-or second-generation members or unrelated healthy controls.This substitution was also found in 1 out of 4 family members of fourth-generation.Conclusions The nonsense mutation c(1696C→T) in the ATP2C1 gene,is likely to be responsible for BFCP in this Chinese four-generation pedigree.The underage family member of fourth-generation who carried the mutation c(1696C→T) but had no clinical symptoms of BFCP,should be closely followed.

Objective To investigate the regulation and the mechanism of Melatonin in calcium overload of pancreatic acinar cell in acute necrotizing pancreatitis (ANP). Methods Fifty-four Sprague-Dawley (SD) rats were equally divided into three groups:sham-operation group (SO group),ANP group (created with retrograde cholangiopancreatography injection of sodium taurocholate) and MT group (ANP model made after intra-peritoneal injection MT for 30 mins).Rats were sacrificed at 1,4 and 8hours after operation and pancreas tissues were underwent pathological examination.The free calcium concentration of pancreas tissues was determined by fluorescence minitoring method; and the expression of CaMK Ⅱ in pancreas tissues at mRNA and protein level was tested by real-time PCR and Western Blot.Results Pancreatic pathological injury in ANP groups was progressively severe as time extended,which was obviously ameliorated in MT group compared with ANP group (the t value of compared pathological score at 1,4 and 8 hour was:-7.95,-9.72 and -7.69,all P=0.00).Compared at same time point,the free Ca2+ concentration of pancreas tissues in ANP group was significantly higher than that of SO group (the t value of 1,4 and 8 hour was 13.09,18.58 and 16.56,all P=0.00).It was a little bit higher in MT group compared with that of SO group,however was significantly lower than that of ANP group (the t value of 1,4 and 8 hour was -10.03,-11.75 and -11.02,all P =0.00).Compared with SO group,the expression of CaMK Ⅱ at mRNA and protein level significantly increased in ANP group; MT significantly inhibited its expression.Conclusions The expression of CaMK Ⅱ may be inhibited by MT interfere,and then lower the calcium overload in pancreatic acinar cell,which play a role in pancreas tissue protection.

Objective To predict the CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains, which were prevailing in China. Methods Total of 236 plasma samples were from the 3rd National HIV Molecular Epidemic Survey (NMES3). All the subjects were infected with HIV-1 CRF07_BC viruses. The tat exon 1 region was amplified by reverse transcription reaction and nested polymerase chain reaction (nested-PCR), then the PCR products were sequenced. The distribution of CTL epitopes of this region were predicted by on-line software BIMAS HLA Peptide Binding Predictions and statistics software. Results To-tal of 236 CRF07_BC strains were from 16 provinces, mainly in intravenous drug asers(58.9%)and then sex(25.0%). It was showed that there were 12 CTL epitopes of 236 Tat exon 1 region of CRF07_BC strains mainly located in proline-rich region, cysteine-rich region and core-region. Those epitopes were banded by 5 HLA presenting molecules in genotype(A * 2501 ,A * 2902, B * 15,B * 5301 and Cw * 1203) and 6 HLA presenting molecules in serotype (B53, B58 ,B57 ,A3 ,A68 and Cw12). The frequency of single amino acid substitution was more than 50% in 7 CTL epitopes. Conclusion The CTL epitopes in Tat exon 1 of CRF07 _BC strains were located in different functional regions, and there were some amino acid variations in them.

Objective To investigate the safety and efficacy of three column osteotomy (3-CO) procedures through previous spinal fusion site for the revision surgical treatment in severe spinal deformity patients.Methods From Oct.2010 to May 2014 in our hospital,a total of 12 severe spinal deformity patients underwent 3-CO for the revision surgical treatment.There were 7 males and 5 females with the average age of (21.8±3.8) years,ranging from 18 to 30 years.The mean time from the initial operation to the revision surgery was (10.2±4.8) years (ranging from 3 to 17 years).The reasons for revision were:curve progression in 7 patients,neurologic deficit in 2 cases,implant failure in 1 patient and pseudarthrosis in 2 patients.The coronal parameters including major Cobb angle and distance between C7 plumb line and center sacral vertical line (C7PL-CSVL),and the sagittal parameters including global kyphosis curve and sagittal vertical axis (SVA) were measured pre-operatively,post-operatively and at last followup,respectively.The operation time,intraoperative blood loss and complications were recorded.The paired t test was used to evaluate the difference among pre-revision,post-revision and last follow-up.Results The average operation time was (451.7±83.1) min (range,320-600 min) and the average blood loss was (4 016.7± 1 080.0) ml (range,2 700-6 000 ml).The average follow-up time after revision operation was (35.4±9.8) months (range,24-49 months).The coronal Cobb angles of pre-revision and post-revision were 83.8°±23.3°and 34.6°± 13.7°.The average correction rate was 60.1％ ±8.8％.At last follow-up,the average coronal Cobb angle was 34.9°±13.8°,there was no significant loss of correction.The pre-revision and post-revision values of global kyphosis were 99.1°±13.3°and 38.7°±7.8° with a mean correction rate of 60.8％ ±6.7％.At the last follow-up,the average global kyphosis was 39.3°±7.5°and no loss of correction was found.For the C7PL-CSVL and SVA,pre-revision (30.3± 17.1) mm and (40.1±31.1) mm were corrected to (14.3 ±7.6) mm and (19.1± 12.3) mm immediately after revision operation,respectively.At final follow-up,the average C7PL-CSVL and SVA were(14.1 ± 7.6) mm and (19.6± 12.1) mm,the correction was well maintained.Obviously,two patient's neurological status improved from Frankel C before revision surgery to Frankel E.Complications were encountered in five patients (41.7％),including SEP signal changed in 1 patient (8.3％),transient neurologic deficit after revision surgery in 1 patient (8.3％),cerebrospinal fluid leak in 1 patient (8.3％),and pleural effusion in 2 patients (16.7％).During the follow-up time,there was no patient experienced pseudarthrosis,implant failure,infection or significant loss of correction.Conclusion Based on results of this study,it was concluded that 3-CO procedures through previous spinal fusion sites could obtain satisfactory and safety results in severe spinal deformity revision surgery.However,it is a technique-demanding procedure with more blood loss,longer operative time and higher risk of perioperative complications.

ObjectiveTo investigate the role of platelet-derived growth factor(PDGF) and its receptor in the development of hypertrophic scar. MethodsThe expression of PDGF and its receptor were detected in biopsy specimens of 9 pieces of normal skin, 7 pieces of granulation tissue of burn wound and 34 pieces of hypertrophic scar by immunohistochemical staining using specific polyclonal antibodies.ResultsPDGF and its receptor markedly increased in granulation tissue and hypertrophic scars, reaching the peak in the hypertrophic scars within 6 months and then decreased after the peak, whereas PDGF and its receptor expressed weakly in only a few normal skin specimens, and the differences were significant(P<0.05).ConclusionsThe increasing expression of PDGF and its receptor may be related to the development of hypertrophic scar.

OBJECTIVETo study the mechanism of the cellular proteins involved in the process of replication of hepatitis C virus (HCV) negative-strand RNA.

METHODSUltraviolet (UV) cross-linking was used to identify the cellular proteins that would bind to the 3'-end of HCV negative-strand RNA. Competition experiment was used to confirm the specificity of this binding, in which excess nonhomologous protein and RNA transcripts were used as competitors. The required binding sequence was determined by mapping, then the binding site was predicted through secondary structure analysis.

RESULTSA cellular protein of 45 kD (p45) was found to bind specifically to the 3'-end of HCV negative-strand RNA by UV cross-linking. Nonhomologous proteins and RNA transcripts could not compete out this binding, whereas the unlabeled 3'-end of HCV negative-strand RNA could. Mapping of the protein-binding site suggested that the 3'-end 131-278nt of HCV negative-strand RNA was the possible protein-binding region. Analysis of RNA secondary structure presumed that the potential binding site was located at 194-GAAAGAAC-201.

CONCLUSIONThe cellular protein p45 could specifically bind to the secondary structure of the 3'-end of HCV intermediate negative-strand RNA, and may play an important role in HCV RNA replication.

Binding Sites ; Hepacivirus ; genetics ; Nucleic Acid Conformation ; RNA, Viral ; chemistry ; metabolism ; RNA-Binding Proteins ; analysis ; metabolism ; Virus Replication

Objective:To investigate the diagnostic values of interleukin-22 (IL-22), interferon-γ(IFN-γ)and macrophage migration inhibition factor (MIF) in pleural effusion for tuberculosis pleurisy.Methods:From April 2018 to May 2019, a total of 77 patients including 45 cases of tuberculous pleurisy, 19 cases of malignant pleurisy, 13 cases of parapneumonia and 13 cases of healthy control in Wuxi Fifth People′s Hospital were enrolled. The levels of IL-22, IFN-γ and MIF in plasma and pleural effusion were detected by enzyme linked immunosorbent assay (ELISA). Mann-Whitney U test was used for statistical analysis.The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic values of IL-22, IFN-γ and MIF for tuberculous pleurisy. Results:The median levels of IL-22, IFN-γ, MIF and adenosine deaminase in 45 cases with pleural effusion in tuberculosis pleurisy group were 396.8 ng/L, 2 200.0 ng/L, 241.3 μg/L and 70.8 U/L, respectively, which were all significantly higher than 32 cases with non-tuberculosis pleurisy group, including 19 cases with malignant pleurisy and 13 cases with parapneumonia (52.8 ng/L, 232.3 ng/L, 179.6 μg/L and 17.0 U/L, respectively). The differences were all statistically significant ( U=179.000, 118.500, 287.000, 162.000, respectively, all P<0.05). The median levels of IL-22 and IFN-γ in plasma of tuberculosis pleurisy group were 20.0 ng/L and 45.9 ng/L, respectively, which were both higher than healthy control group (14.3 ng/L and 33.4 ng/L, respectively). The level of MIF was 96.2 μg/L, which was lower than healthy control (159.5 μg/L). The differences were all statistically significant ( U=74.000, 13.000 and 73.000, respectively, all P<0.05). The areas under ROC curve (AUC) of IL-22, IFN-γ and MIF in pleural effusion for the diagnosis of tuberculosis pleurisy were 0.876, 0.917 and 0.682, respectively.The sensitivities were 93.75%, 100.00% and 63.64%, respectively; the specificities were 82.22%, 91.11% and 65.85%, respectively. The median levels of IL-22 and IFN-γ in plasma in tuberculosis pleurisy group at two months of follow-up after anti-tuberculosis therapy were 16.0 ng/L and 33.9 ng/L, respectively, which were both lower than baseline (20.0 ng/L and 44.7 ng/L, respectively). The differences were both statistically significant ( U=2.156 and 2.221, respectively, both P<0.05). Conclusion:IFN-γ and IL-22 in pleural effusion could be used as effective indicators to identify tuberculous pleurisy, and the dynamic monitoring of IL-22 in patients′plasma could be an important biomarker in evaluating the efficacy of anti-tuberculosis treatment.

Pancreatic duct stone is a rare pancreatic disease in clinic, which is often associated with chronic pancreatitis, and could seriously damage the quality of life of patients, and even induce pancreatic cancer. The diagnosis is mainly based on imaging examination, and the treatment methods are diverse. It is necessary to follow the principle of individualized treatment and treat it as soon as possible. This article reviewed the etiology, mechanism, diagnosis, classification and treatment of the disease.