ObjectiveTo determine the prevalence of avian influenza viruses in the external environment of poultry in Anqing City， Anhui Province， and provide scientific evidence for prevention and control of animal-derived influenza in humans. MethodsA total of 28 farmers’ markets/farms in 10 counties （cities， districts） of Anqing City， Anhui Province， were selected as surveillance sites by simple random sampling strategy. Poultry faeces and other related samples were collected for 6 consecutive weeks. Real-time fluorescence quantitative PCR was used to examine the nucleic acids of influenza A virus. Subtypes H5， H7， and H9 of avian influenza virus were further tested in the positive samples. ResultsA total of 426 specimens were collected， among which 113 tested positive with a positive rate of 26.53%. Among the positive specimens， 104 were determined to be subtype H9， accounting for 92.04%. It did not significantly differ in the positive rate between the main and non-main urban areas （χ²<0.01， P>0.05） or among the specimens collected in different weeks （χ²=7.57， P>0.05）. However， it significantly differed in the positive rate among the specimens collected in the third week and other weeks （χ²=6.89， P<0.05）. Furthermore， among the different sampling sites， farms had the highest positive rate of 46.67%. Among the specimens from different sources， the surface-coated specimens from poultry cages had the highest positive rate of 34.78%. ConclusionAvian influenza viruses are prevalent in the external environment of poultry in Anqing City. It warrants strengthening the surveillance and risk assessment to reduce the virus transmission in the external environment and risk of human infection with animal-derived influenza.

Antibodies, Neutralizing ; Antibodies, Viral/isolation & purification* ; B-Lymphocytes/virology* ; COVID-19/virology* ; Humans ; Immunity/genetics* ; RNA, Viral/immunology* ; SARS-CoV-2/pathogenicity* ; Single-Cell Analysis

Objective To observe the influence of IL-12,IL-15 on CIK cell in the normal culture;to observe the anti-tumor effect in the circumstance of different combination of cytokines,and to provide a new insight for preparing high effective and qualified CIK cell in vitro.Methods The optimal concentrations of IL-2,IL-12 and IL-15 were determined,respectively.After the peripheral blood from healthy blood donors was collected,monocytes were selected and co-cultured with different cytokines into different groups,as group A(IL-2 normal culture group),group B(IL-2 and IL-12 group),group C(IL-2 and IL-15 group),group D(IL-2,IL-12 and IL-15 group),and group E(cytokine control group).The monocytes in different groups were calculated by globulimeter,the activity of cells was detected by Trypan blue staining,positive ratio of CD3,CD8,CD56 on the celluar membrane was detected by flow cytometry,and the anti-tumor effect of CIK to SMMC-7721 was detected by MTSmethod,inthedayof0,5,10,15,20 after the culture.Results Statistical analysis indicated that,the proliferation multiplication of CIK cells was significantly higher in group B,group C and group D after 10,15 and 20 days of culture than those in group A(P＜0.05);and group D had higher proliferation multi-plication than that of group C(P＜0.05).The percentage of CD3 + CD8 +,CD3 + CD56+ in CIK cell membrane in group B,C,D was significantly higher than that in group A after 15 and 20 days of culture (P＜0.05).The percentage of CD3+ CD8+,CD3+ CD56+ in CIK cell membrane in group D was significantly higher than that in group B after 15 and 20 days of culture (P＜0.05).The killing rate of CIK cells for liver cancer in each group at 10,15,20 days of culture was significantly higher than that of group A when the target target ratio was 5 ∶ 1 (P＜0.05).The killing rate of CIK cells for liver cancer in group D,C at 10,15,20 days of culture was significantly higher than that of group B(P＜0.05).Conclusion IL-12and IL-15 could improve the proliferation of CIK cells,and IL-15 also has the effect of enhancing CIK cells the tumor-killing to SMMC-7721 activity.

Objective To analyze the clinical and imaging characteristics of pediatric neuromyelitis optica spectrum disorders(NMOSD)in children. Methods The clinical data,imaging manifestations and follow - up data of 16 NMOSD patients at Department of Pediatric Neurology,Shandong Provincial Hospital Affiliated to Shandong Univer-sity between July 2013 and September 2017 were respectively analyzed. Results In 16 patients,initial presentations included optica neuritis(ON)in 5 cases,longitudinally extensive transverse myelitis(LETM)in 6 cases,and among them there were 2 cases with acute disseminated encephalomyelitis and 3 cases with both ON and LETM. Eleven cases received aquaporin - 4(AQP4)antibody examination and 4 cases were found seropositive. One case out of 7 detected cases was found AQP4 antibody positive in cerebrospinal fluid. Eleven cases received optica magnetic resonance imaging (MRI),and 8 cases were found abnormal signals in optic nerve and optica chiasma. The spinal cord MRI showed 13 ca-ses with LETM manifestations,and abnormal signals were found in vertebral segments(5 - 13),and among them 1 case had cervical cord,3 cases were thoracic cord and 9 cases were both of the above. Lesions in the cervical cord in 2 cases were extended upward to the medulla. Fifteen cases received brain MRI and all of them had brain lesions,which were mainly involved in the central and subcortical white matter,thalamus,corpus callosum,brainstem,the junction of spinal cord and medulla,cerebellum,and so on. All patients received treatment for acute attacks with high - dose Methylpred-nisolone and/ or gamma globulin and got obvious relief. Two cases with recurrent ON received treatment of Rituximab and their vision became improved. Fifteen patients were followed up,and 2 cases had limb disorders and 4 cases had visual impairment,other patients had no clinical symptoms. Conclusions Pediatric NMOSD has a diverse clinical pre-sentation at the onset disease. Those who are initial diagnosed acute myelitis,ON and acute disseminated encephalomye-litis should be considered the possibility of NMOSD. Antibody to AQP4 testing can assist the diagnosis. The typical ima-ging characters of NMOSD children are abnormal signals in the high expression area of AQP4. Intracranial lesions are more common in children. The acute treatment includes the high - dose Methylprednisolone and gamma globulin. Rituximab can be used for the recurrent patients.

Objective To measure the expression of indoleamine 2, 3-dioxygenase(IDO)in condy-loma acuminatum (CA) lesions, and to evaluate its ability to locally metabolize tryptophan. Methods Immunohistochemical study was performed to observe the protein expression of IDO in skin lesions of patients with CA, and count the number of IDO-positive cells. Immunofluorescence assay was conducted to estimate the relationship between IDO-positive cells and dendritic cells. Epidermal cells and keratinocytes were isolated from warts of 30 patients with CA and prepuces of 11 healthy controls respectively, and both in vitro incubated with tryptophan solution for 4 hours. Then, high-performance liquid chromatography (HPLC)was performed to detect the level of tryptophan metabolite, kynurenine, in the culture supernatant of the above cells, which could reflect the ability of epidermal cells to metabolize tryptophan. Results Rare IDO-positive cells were found in the normal skin, but a lot of IDO-positive cells gathered in the epidermis of the wart tissues. The IDO-positive cell/total cell ratio was significantly higher in the wart tissues than in the normal skin(48.3%± 15.4%vs. 5.2%± 2.4%, P<0.05). The fluorescence signals of IDO-positive cells and CD1a-positive Langerhans cells were not overlapped with each other, suggesting that IDO-positive cells were derived from epidermal cells of the wart tissues. Compared with the keratinocytes from the healthy skin, the epidermal cells from warts had a stronger ability to metabolize tryptophan in vitro. Conclusion A large number of IDO-positive cells exist in CA warts, and may be involved in occurrence of CA.

Objective To investigate the clinical efficacy of rectal resection for rectal cancer.Methods 35 patients with rectal cancer were diagnosed by pathological examination.They were treated with laparoscopic rectal cancer resection and conventional chemotherapy.The patients were followed up for 3 years.The operative time,blood loss,intraoperative and postoperative complications,postoperative recovery,death and so on were observed.Results The patients were operated successfully,and no tumor cells were found in the bowel edge.The average operative time was (171.74 ± 58.24) min,average blood loss was (85.74 ± 68.32) ml,there were no infection,bleeding,anastomotic complications.After 2 ～ 3 years of follow-up,there was 1 patient with liver metastases,and no local recurrence,no fecal incontinence and no deaths.Conclusion Rectal resection for rectal cancer had good effect and could improve patients' quality of life.

To report a case of cutaneous and subcutaneous coinfection caused by Lichtheimia corymbifera and Candida parapsilosis.A 67-year-old female peasant consulted about proliferative granuloma developing on her left forearm after topical application of a Chinese herbal drug and splint fixation for the treatment of suspected fracture of the wrist.Direct microscopic examination showed gram positive budding yeast cells in lesion secretions.Pathological study with periodic acid-Schiff (PAS) and gormori methenamine silver (GMS) staining revealed broad non-separate hyphae in the corneum and dermis.Fungal culture of lesional tissue at 35℃ grew both mould and yeast.The mould was identified as Lichtheimia corymbifera based on morphological findings and sequences of the internal transcribed space (ITS) 1-4 regions.Thermal tolerance study revealed that the isolate grew fast at 37℃ but slowly at 40℃.Under a scanning electron microscope,the acrogenous sporangia were pear-shaped with conical sporangiophores originating from the top of stolon,which were among but not opposite to the rhizoids.The yeast was identified as Candida parapsilosis by Chromagar test and D1/D2 region sequencing.As antimicrobial susceptibility test indicated,the Lichtheimia corymbifera isolate was most sensitive to terbinafine and itraconazole.The proteolytic activity of Lichtheimia corymbifera was higher than that of Candida parapsilosis.The granuloma completely subsided after surgical resection and 6-week treatment with oral itraconazole 200 mg twice a day.No recurrence was observed during a 4-year follow-up.

Objective To observe the effect of computer anorectal therapeutic apparatus in treatmenf of hemorrhoids. Methods 100 cases of hemorrhoids were selected and randomly divided into A group and B group,each group 50 cases. The patients of A group were treated by computer anorectal therapeutic equipment. The patients of B group were administrated with traditional surgical treatment. Then the pain, urine retention, wound edema, the occurrence of postoperative bleeding, wound healing time and the average following - up outcome were observed within 3 days after operation. Results A group had occur pain, urine retention, wound bleeding and wound edema in cases of 3,1,1,8 respectively, and B group had them in cases of 7,3,5,13 respectively,there was no significant difference between the two groups(all P ＜0. 05). The healing time of A group and B group was (13. 5 ±4.6) days and (14.3 ± 4.9) days respectively, there was no significant difference between the two groups (P ＞ 0. 05). 46 cases of A group were cured , 42 cases of B group were cured, there was no significant difference between the two groups (P ＞ 0.05). Conclusion Computer anorectal therapeutic apparatus in treatmenf of hemorrhoids could reduce hemorrhoids bleeding, pain, urine retention ,edema and wound complications, and had good effect.

Objective To partially purify the toxic factor secreted by A. corymbifera and to analyze the mechanism of A. corymbifera-induced human umbilical vein endothelial cell (HUVEC) apoptosis. Methods Glycoprotein secreted by A. corymbifera was purified by Con A Lectin chromatography. The influence of different protein fractions on HUVEC apoptosis was determined by flow eytometer. Both denaturing and nondenaturing deglycosylation of purified glycoprotein was performed and the ability of the protein moiety and carbohydrate moiety to induce HUVEC apoptosis was evaluated respectively. Activation of related caspases during A. corymbifera-induced apoptosis was analyzed by Western blot. The role of caspase-8 and -9 in HUVEC apoptosis was investigated using caspase inhibitors. Caspase inhibitors were used to stop the suppression of HUVEC viability by XTT assay. Results Flow cytometric analysis shows the total protein as well as the glycoprotein fraction of A. corymbifera may induce HUVEC apoptosis in a dose dependent manner. In contrast, similar activity was not observed in the non-glycoprotein fraction. Neither deglycosylated protein nor carbohydrate moiety is able to induce HUVEC apoptosis alone. In the apoptotic signaling pathway, caspase9, caspase-3 and cytochrome C were activated significantly, except caspase-8. Moreover, caspase-9 inhibitor, instead of caspase-8 inhibitor, completely abrogates A. corymbifera-induced HUVEC apoptosis. Caspase9 and caspase-3 inhibitors completely waived the suppression of HUVEC viability by A. corymbifera. Conclusion Glycoprotein secreted by A. corymbifera is associated with HUVEC apoptosis. Intact glycoprotein is essential for the apoptotic progress. Intrinsic apoptotic signaling pathway mediates A. corymbifera-induced HUVEC apoptosis.

Objective To analyze the influence of Absidia corymbifera on cell activity of human umbilical vein endothelial cells (HUVEC) as well as the related mechanism. Methods Time course analy sis of the influence of A. corymbifera on cell viability of HUVEC was determined by cell counting after Trypan blue staining. Apoptosis of HUVEC induced by A. corymbifera was observed under fluorescence microscope after treatment with apoptosis detection kit. Time course analysis of HUVEC apoptosis induced by A. corymbifera was detected by flow cytometry quantitatively. Effect of caspase-3 inhibitor on A. corymbifera associated apoptosis was also evaluated at the same time. Activation of caspase-3 inside HUVEC was detected by Western blot. Results A. corymbifera inhibited cell viability of HUVEC in a time-dependent manner by Trypan blue staining. After 12 hours' co-culture, A. corymbifera began to show suppression on cell viability (P =0. 001 ). Fluorescence microscope observation revealed A. corymbifera induced apoptosis of HUVEC instead of necrosis. Flow cytometry analysis showed A. corymbifera induced apoptosis of HUVEC in a time-dependent manner. A. corymbifera began to show obvious effect on apoptosis after 12 h co-culture (P =0.0036). Moreover, A. corymbifera-associated apoptosis was almost abrogated completely by caspase-3 inhibitor. Western blot analysis demonstrated that A. corymbifera triggered the activation of caspase-3 inside HUVEC in a timedependent fashion. Conclusion A. corymbifera induces apoptosis of HUVEC in vitro. Such apoptotic signal is transmitted through caspase cascade reaction.