Effect of hTERC and FOXP3 gene magnification in condyloma acuminate lesion on the immune response and cell apoptosis

Objective:To study the effect of hTERC and FOXP3 gene amplification in condyloma acuminate lesions on the immune response and cell apoptosis.Methods:The condyloma acuminate lesions diagnosed between May 2014 and October 2016 and the normal skin tissue from circumcision during the same period were collected to extract RNA and then determine hTERC, FOXP3 and apoptosis-related gene mRNA amplification, and after protein extraction, the protein levels of immune response-related cytokines were determined.Results:hTERC, FOXP3, Livin and Survivin gene mRNA amplification as well as IL-4 and IL-10 protein levels in condyloma acuminatum lesions were significantly higher than those in normal skin tissue while TRAIL, Caspase-3 and PDCD4 mRNA amplification as well as IL-2 and TNF-α protein levels were significantly lower than those in normal skin tissue;IL-2 and TNF-α protein levels in condyloma acuminatum lesions with high FOXP3 mRNA expression were significantly lower than those in condyloma acuminatum lesions with low FOXP3 mRNA expression while IL-4 and IL-10 protein levels were significantly higher than those in condyloma acuminatum lesions with low FOXP3 mRNA expression;Livin and Survivin mRNA amplification in condyloma acuminatum lesions with high hTERC mRNA expression were significantly higher than those in condyloma acuminatum lesions with low hTERC mRNA expression while TRAIL, Caspase-3 and PDCD4 mRNA amplification were significantly lower than those in condyloma acuminatum lesions with low hTERC mRNA expression.Conclusion:Highly expressed hTERC and FOXP3 genes in condyloma acuminatum lesions can inhibit apoptosis and inhibit antiviral immune response respectively.

The Condyloma Acuminatum’s Recurrence Can Be Reduced by Lesional Auto-Transplantation

Background: Condyloma acuminatum (CA), also known as a genital wart (GW), is a sexually transmitted disease caused by human papillomavirus (HPV). HPV lesions, recurrence tendency, and risk of malignant formation are primarily dependent on the person’s immunity level. GW recurrence is a major challenge in CA treatment. The aim of this study was to explore how lesional auto-transplantation (LT) can be used to treat CA and decrease its recurrence. Methods: We treated CA through the preparation and implantation of tissue from nine CA patients in our dermatology clinic. Transplantation of small pieces of HPV lesions to the subcutaneous fat of the inguinal region was carried out with the help of a simple surgical method under local anesthesia. Patients were followed up for six to eight months. We searched PubMed and the Web of Science for treatment options for CA to compare our treatment method recurrence rate with existing treatment options. Results: During three months of follow-up, seven patients experienced no recurrence of condyloma lesions, while two patients experienced recurrence of small lesions of condyloma acuminatum. The recurrence rate of CA once treated by auto-transplantation was relatively low compared to other CA treatment options. Conclusions: Transplantation of HPV lesions to the inguinal area reduces the recurrence rate in sexually active individuals. The 2/9 recurrence after implantation could be due to poor hygiene, sexual relationships or immune factors.

DIFFERENTIATION OF PSEUDOCONDYLOMA OF VULVA AND CONDYLOMA ACUMINATA BY DOT BLOT HYBRIDIZATION AND POLYMERASE CHAIN REACTION

This study differentiated pseudocondyloma of vulva from condyloma acuminata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases of pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the study. The genital lesions were examined clinically and were biopsied. Each biopsy was subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82. 6%) out of 23 cases of condyloma acuminata and 2(25% ) out of 8 cases pseudocondyloma of vulvae (P<0. 05). PCR detected HPV DNA in 51 (92. 7%) out of 55 cases of condyloma acuminata , compared with none in 23 cases of pseudocondyloma (P<0.001 ). HPV DNA was present in the majority of condyloma acuminata specimens. HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful technique for HPV DNA detection.

Intracellular Cytokine Expression Profile in Patients with Condyloma Acuminata

Objective: To study the immunological mechanisms ofCondyloma acuminata(CA) through investigating Tlymphocyte subset levels and cytokine profile in theperipheral blood of patients with CondylomaAcuminata. Methods: Tricolor and bicolor immunofluorescentstaining antibody of cell surface antigen and intracel-lular IL-2, IL-4, IL-12, IFN-Y in CD4^+ and CD8^+ T-lymphocytes from 20 patients with CA were performedand followed by flow cytometry. Results: The number of CD3^+T, CD4^+T-lymphocytescells and CD4^+/CD8^+T cells ratio were significantlydecreased(P<0.01) in patients with CA Compared tocontrols, and IL-2, IL-12, IFN-γ production in CD4^+Tcells was decreased(P<0.01), IL-4 and IFN-γ produc-tion in CD4^+T cells was not significantly different (P>0.05), while IL-2 and IL-12 production in CD8^+Tc cellswas decreased (P<0.01), whereas IFN-γ and IL-4 pro-ducing in CD4^+T cells were of no significantlydifference(P>0.05). Conclusions: There was an imbalance of T lympho-cyte subsets, Th1/Th2 cytokines and Tc1/Tc2 in theperipheral blood of CA patients, which may play animportant role in the pathogenesis and progression ofCA.

Observation of Ultrastructure in Lesions of Condyloma Acuminatum

Objective: To explore the effect of morphological changes in the development of condyloma acuminatum (CA) Materials and Methods: Lesions in five patients with CA were observed . Results: Upon electron microscopy, the most characteristic feature of the lesions important for diagnosis, was the presence of distinct perinuclear vacuolizations, or so-called koilocytes, among some epithelial cells. These cells possessed hyperchromatic nuclei, swollen mitochondria, dilated endoplasmic reticulum and dissolved glycogen. There were interchromatin granules and perichromatin granules in some nuclei. Moreover some virus particles were also seen in the nuclei of some infected cells. Conclusions: The ultrastructural findings may be used to histopathologically explain the pathogenesis and mechanism of this disease, and it is helpful for diagnosis of CA .

Fas and Bc1-2 EXpression on T Lymphocyte Subsets in the Peripheral Blood of Relapsing Patients with Condyloma Acuminatum

Objective: To study the expression of Fas and Bcl-2proteins on T lymphocyte subsets in the peripheralblood of relapsing patients with condyloma acuminatum(CA) and healthy controls. Methods: Flow cytometry (permeabization andstaining procedure with conjugated antibodies) wasused. Results: We observed that the expression of Fasprotein on CD4^+ T lymphocyte subset of CA patientswas significantly higher than that of healthy controls(P<0.01). Conclusions: Increased expression of Fas proteinon CD4^+ T lymphocyte subset may be a cause of de-creased percentage of CD4^+ T lymphocyte subset. Thisinduces the increased ratio of CD4^+/CD8^+.

Effect of Activation-induced Cell Death of Peripheral Blood Mononuclear cells in Patients with Condyloma Acuminatum

Objective: To investigate the effect of activation-induced cell death (AICD) on cellular immune function in the condyloma acuminatum(CA). Methods: Peripheral blood mononuclear cells (PBMC) were isolated from normal healthy individuals (control group) and patients with CA. PBMC were cultured with PHA-P for 48h in vitro. Apoptosis of the PBMC was detected by flow cytometry. Supernatant cytokines (IL-2 and IL-10) were assayed by ELISA. Results: The rate of PBMC apoptosis in both CA group and control group in fresh PBMC was very low and similar in both groups(P>0.05). The rate of PBMC apoptosis within the CA group was noticeably increased compared to that of the control (P<0.001)af-ter PBMC were cultured for 48h. The level of IL-2 was significantly lower in the CA group than in the control group (P<0.001), The level of IL-10 was significantly higher in the CA group compared to thecontrol group(P<0.001). Conclusion: Study results indicate that AICD may affect cellular mediated immune function and play an important role in the pathogenesis of CA.

Condyloma acuminatum of the anal canal,treated with endoscopic submucosal dissection

Condyloma acuminatum(CA) is a common sexually transmitted disease caused by human papilloma virus infection. Not all individuals develop persistent, progressive disease, but careful follow up is required with moderate-to-severe dysplasia to prevent progression to malignancy. Standard therapies include surgical treatments(trans-anal resection and transanal endoscopic microsurgery) and immunotherapeutic and topical methods(topical imiquimod); however, local recurrence remains a considerable problem. Here, we report a case with superficial CA of the anal canal, treated with endoscopic submucosal dissection(ESD). A 28-year-old man presented with a chief complaint of hematochezia. Digital exam did not detect a tumor. Screening colonoscopy revealed 10-mm long, whitish condyles extending from the anal canal to the lower rectum. The lesion covered almost the whole circumference, and only a small amount of normal mucosa remained. Magnifying endoscopy with narrow band imaging showed brownish hairpin-shaped, coiled capillaries. Although histopathological diagnosis by biopsy revealed CA, accurate histological differentiation between CA, papilloma, and squamous cell carcinoma can be difficult with a small specimen. Therefore, weperformed diagnostic ESD, which provides a complete specimen for precise histopathological evaluation. The pathological diagnosis was CA, with moderate dysplasia(anal intraepithelial neoplasia 2). There was no recurrence at 16 mo after the initial ESD. Compared to surgical treatment, endoscopic diagnosis and resection could be performed simultaneously and the tumor margin observed clearly with a magnifying chromocolonoscopy, resulting in less recurrence. These findings suggest that endoscopic resection may be an alternative method for CA that prevents recurrence.

喜树(Camptotheca acuminate)幼苗形态和光合作用对不同光环境的适应

选择旷地、林隙、林下3种光环境,研究了喜树幼苗总生物量、生物量分配、株高、基茎、叶面积和光合作用的差异．结果表明:①旷地中幼苗的平均相对生长速率最大,总生物量累积最大,地下部分生物量分配明显增多; 林隙中叶生物量比显著增加;林下茎生物量比显著增加．②在林下和林隙环境中,喜树幼苗增加高度以截获更多的光能．比叶面积在林隙、林下显著高于旷地．③3种光环境中,净光合速率日变化均呈单峰型,约在中午12:00 时达到峰值,旷地中日均净光合速率大于林隙、林下．④林隙和林下光补偿点(LCP)降低适应低光环境,光饱和点 (LSP)在各环境中差异不大．喜树幼苗能通过不同的形态和生理特性的变化适应各光环境．

Correlation between the Expression of Fas, Bcl-2 in Peripheral Blood Lymphocytes and the LeveL of IFN-γ,IL-4 in Serum of Patients with Condyloma Acuminata

In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Bcl 2) in peripheral blood lymphocytes (PBLC) and the level of IFN γ and IL 4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl 2 in PBLC and the level of IFN γ and IL 4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl 2 was significantly lower (both P< 0.01). The level of IFN γ in serum of CA was significantly lower than in the normal control group ( P< 0.01), but IL 4 was significantly lower (both P< 0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN γ in serum of patients with CA, but had a positive correlation with the level of IL 4; The expression of Bcl 2 had a positive correlation with the level of IFN γ, but had a negative correlation with the level of IL 4. All the correlation coefficients had significant differerce by t test ( P< 0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the THl associated cytokines (eg: IFN γ) and the increased secretion of the TH2 asssociated cytokines (eg: IL 4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.

The Lymphoblastoid Cell Lines of Recurrence Condyloma Acuminatum Patients Produce Lower Level of Tumor Necrosis Factor Stimulated with LPS

To study the mechanism of Condyloma acuminatum (CA) recurrence, and the association of CA recurrence with the ability of the host derived lymphoblastoid cell lines (LCL) stimulated by LPS to produce tumor necrosis factor (TNF), EBV-transformed B LCL were used as TNF producing cells The ability of LCL stimulated by LPS to produce TNF was measured by bioassay The results showed that the LCL from CA patients (including recurrent and non-recurrent CA patients) produced similar level of TNF stimulated by LPS to that of normal controls (29 54%±11 28% vs 34 31%±11 46%, P =0 1498) The LCL of CA recurrent patients produced significantly lower amount of TNF than that of non-recurrent CA patients (23 72%±7 41% vs 37 33%±11 10%, P =0 0032) Compared with the normal controls, CA recurrent patients showed a decreased ability to produce TNF (23 72%±7 41 vs 34 31%±11 46, P =0 0054), whereas CA non recurrent patients had the similar ability to the controls (37 33%±11 10 vs 34 31%±11 46, P =0 4914) It was concluded that the onset of CA was not relevant to the individual's ability to produce TNF But the recurrence of CA was associated with the ability to produce TNF It was also indicated that the TNF involved cellular immunity might play an important role in the clearance of the residual HPV by the host after treatment

Telomerase Activity in Condyloma Acuminatum Tissue with Different HPV Types

Summary: The telomerse activity in condyloma acuminatum (CA) tissue with human papillomavirus (HPV) types of 6/11 and 16/18 was detected to investigate the function of telomerase in the occurrence, development and carcinogenesis of genital CA. Forty-two biopsies from patients with gennital CA and 30 control tissue samples were tested for telomerase activity, HPV presence and types. The telomerase activity was determined by modified telomerase repeat amplification protocol (TRAP) assay and HPV typing by polymerase chain reaction (PCR) with typing-specific primers. Results showed that HPV-DNA was negative and the expression rate of telomerase was 16.7 % in all normal skin samples. All CA samples were positive for HPV (6/11 type was found in 32 cases, 16/18 in 3 and mixed type in 7). Telomerase activity was detectable in all CA patients. The telomerase activity in CA of 16/18 type was apparently higher than in CA of 6/11 type. It was concluded that the hyperplasia in CA might be increased as a result of HPV infection, suggesting that the activation of telomerase by HPV, especially by 16/18 type may play a role in the etiology and carcinogenesis of genital CA.

Anal carcinoma in giant anal condyloma, multidisciplinary approach necessary for optimal outcome:Two case reports and review of literature

BACKGROUND Anal cancers are caused by human papilloma virus(HPV). Buschke-Lowenstein tumor also known as giant anal condyloma(GCA) is a variant of giant neglected anal tumors arising from warts caused by HPV infection. HPV are a family of double-stranded DNA viruses and primarily cause sexually transmitted disease of the genitalia and oropharyngeal mucosa. These tumors are slow growing;locally destructive large verrucous masses.CASE SUMMARY We present a series of two cases with large anal tumors harboring invasive cancers and highlight their presentation and management. Tumors with high risk HPV subtypes(HPV 16, 18, 31, 33) may progress into invasive squamous cell carcinoma(SCC). Untreated GCA can attain enormous size and extend into the pelvic organs and bony structures. Some tumors show malignant degeneration into SCC and are often difficult to diagnose given the large size of the tumors.Complete surgical excision with negative margins is the treatment of choice and necessary to prevent recurrence. This is often not feasible and leaves large surgical wounds with tissue defects with delay in healing and increases postoperative morbidity. Pelvic reconstructive techniques including muscle flaps and grafts are often necessary to close the defects. Human immunodeficiency virus and immunocompromised patients generally do poorly with standard treatments.CONCLUSION A multidisciplinary team of colorectal and plastic surgeons, medical and radiation oncologists along with combination treatment modalities are necessary when malignant transformation occurs in GCA, for optimal outcomes.

小果野蕉(Musa acuminate Colla)花粉母细胞减数分裂过程异常的细胞学观察

香蕉是重要的热带水果,由于雄性不育、单性结实等原因导致其杂交育种困难。小果野蕉(Musaacuminate Colla)是现代香蕉栽培种的祖先之一,在理论研究和应用中有非常重要的地位。前期研究发现小果野蕉存在部分可育花粉,但导致花粉活力下降的原因未知。为了探求导致小果野蕉花粉活力降低的细胞学原因,本研究通过亚历山大红染色法对小果野蕉花粉活力进行检测,通过卡宝品红染色对小果野蕉花粉母细胞减数分裂阶段和小孢子发育阶段进行观察。结果表明:小果野蕉花粉活力为84.33%;小孢子发育阶段未见明显异常;正常四分体比例为81.3%;花粉母细胞减数分裂过程终变期、中期Ⅰ、后期Ⅰ、中期Ⅱ、后期Ⅱ均存在异常,比例分别为8.8%、8.5%、7.6%、10.6%、12.1%。因此,可能是部分花粉母细胞减数分裂异常导致小果野蕉花粉活力的降低。

Expression of CD40 and CD40L on Peripheral Blood Mononuclear Cells in Patients with Condyloma Acuminatum

To observe the expression of CD40/CD40L on peripheral blood mononuclear cells （PBMC） in patients with eondyloma aeuminatum （CA）, flow eytometry was employed to examine the expression of CD40 and CD40L on PMBC in 36 patients with CA and 20 healthy controls. Our results showed that mean level of CD40 expression in CA patients was significantly lower than that in the controls （6.58 %±2.74 % vs 14.81 %±6.12 %, t=5. 703, P〈0.05）; the average level of CD40L in CA patients was also significantly lower than that in the controls （0.73 % ±0.54 % vs 2.67 %±2.43 %, t=3. 532, P〈0.05）. Our resutls suggest that the reduced costimulatory interaction of CD40 and CD40L in CA patients may be one of the important factors responsible for the low cellular immunity.

Expression of MDM2 Protein and mRNA in Condyloma Acuminata

In order to investigate the role of murine double minute 2 (MDM2) in cell proliferation and carcinogenesis in condyloma acuminatum (CA), immunohistochemistry and in situ hybridization were used to detect the expression of MDM2 protein and mRNA in normal skin and skin lesions of CA of vulva. PCR was also used to detect HPV types. The results showed that in 32 observed CA specimens, the expression of MDM2 protein and mRNA was detected in 18(56.25 %)and 22(68.75 %)respectively, while the co-expression of MDM2 protein and mRNA was found in 14. PCR results revealed that HPV6/11 and HPV16/18 subtypes were shown in 28(87.5 %)and 4 (12.5 %)respectively out of 32 CA specimens. Out of the 18 positive specimens expressing MDM2 protein, HPV6/11 subtypes were shown in 15 and HPV16/18 subtypes in 3. In 22 positive specimens expressing MDM2 mRNA, HPV6/11 subtypes were shown in 18 and HPV16/18 subtypes in 4. No expression of MDM2 protein and MDM2 mRNA was observed in normal skin. Our study indicated that the overexpression of MDM2 might be involved in malignant proliferation and carcinogenesis of CA.

The Efficacy of Podophyllin Cautery Added to Surgical Excision for Eradication of Vulvar Condylomata Accuminata;Randomized Controlled Trial

Context: Condyloma acuminatum is a common morbidity caused by human papillomavirus infection. Objective: To compare the recurrence rate after surgical excision with or without podophyllin cautery. Design, Setting, Participants: Sixty women were enrolled in a study that was conducted in Cairo from Jan-2017 to Mar-2018. Interventions: All women received the same preparations. After randomization;in the study group (N = 30), podophyllin cautery was added to surgical removal in the follow-up period. In the control group (N = 30), only surgical removal was used. Main Outcome Measure: The main outcome measure was the estimation of the recurrence of the lesion during the follow-up period. The secondary outcome measures were the incidence of adverse events. Results: Both groups were comparable (p-value > 0.05) with regard to the age and BMI. Recurrence was significantly lower (P = 0.001) in the study group than the control group. In the study group, five cases (17.2%) have recurrence whereas, in the control group, 18 (60.0%) had suffered recurrence. The ARR was 42.8% (CI 95%: 18% - 60.9%) with RR of 0.29 (CI 95%: 0.12% - 0.67%) and NNT2 (CI 95%: 6% - 2%). During the follow-up period, 19 cases (65.5%) of the study group experienced pain versus only eight cases (26.7%) in the control group (p = 0.003). However, the development of scars was less in the study group 7 (24.1%) than in the control group 19 (63.3%), (p = 0.002). Conclusion: Podophyllin cautery, when added to surgical removal, is effective in decreasing the incidence recurrence in cases with condylomata acuminata.

Expression of Activated Epidermal Growth Factor Receptor and Transcription Factor E2F in Condyloma Accuminata

Objective: To study the expression of activated epi-dermal growth factor receptor (EGFR) and transcrip-tion factor E2F (E2F) in Condyloma Accuminata(CA)patients. Methods: Immunofluorescent techniques were usedto investigate the expression of activated EGFR andE2F in CA patients. Results: The expression of activated EGFR on themembrane of epithelial cells in CA lesions was sig-nificantly greater compared to expression levers inthe control group (P<0.01). Moreover, the co-expres-sion of activated EGFR and E2F was significantly in-creased compared to the control group (P<0.01). Conclusion: Our observations suggest that the in-crease in activated EGFR expression may stimulatehyperplasia in CA patients through the activation oftranscription factor E2F.

Changes of Circulating Platelet-Derived Growth Factor and Granulocyte Colony-Stimulating Factor in Patients with Condyloma Acuminatum

Objective: To study the expression levels of platelet-derived growth factor (PDGF) and granulocyte colony-stimulatingfactor (G-CSF) in peripheral blood and their role in thepathogenesis of Condyloma acuminatum (CA). Methods: Sera were taken from 70 patients with Condylomaacuminatum and compared with 35 healthy controls. PDGFand G-CSF in serum were quantitated using a dual antibodysandwich enzyme-linked immunoabsorbent assay (ELISA). Results: Serum concentrations of PDGF and G-CSF weresignificantly increased in patients with Condylomaacuminatum (CA) compared to controls (P<0.001 and P<0.005respectively). Serum levels of PDGF and G-CSF correlatedwith clinical severity of CA, but no significant difference wasobserved between different duration of disease groups. Asignificant positive correlation was noticed between neutrophilcount and G-CSF levels (γ=0.38, P<0.001), and the neutrophilcount showed no significant correlation with PDGE Conclusion: The results indicated that increased expressionof PDGF an -CSF in peripheral blood might be involved in pathogenesis of CA.

The Expression of Fas/FasL in Peripheral Blood Lymphocytes and the Level of IL-2 in Serum of Patients with Condyloma Acuminata

In order to investigate the role of the expression of the Fas/FasL in peripheral blood lymphocytes and the level of IL 2 in serum of patients with condyloma acuminata in the immune pathogenesis of CA, flow cytometry, indirect immunofluorescence labeling and ELISA were performed to detect the expression of the apoptotic regulatory proteins Fas/FasL in PBLC and the level of IL 2 in serum of 60 patients with different course of CA. The results showed that the expression of Fas/FasL in PBLC of the group of short course in CA was significantly higher than that of the normal controls ; the expression of Fas, FasL in PBLC of the group of long course CA was significantly higher than that of the group of short course and the normal controls ( P <0.05 and P <0.01, respectively); the level of IL 2 in serum of the group of short and long course CA was significantly lower than that of the group of normal controls ; the negative relation was revealed between the expression of Fas/FasL in PBLC and the level of IL 2 in serum of patients with CA . It was suggested that the abnormal apoptosis in PBLC and decreased level of IL 2 in serum of CA might play an important role in the course of CA.