In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

Objective:To explore the clinical effects of parallel combined flow-through perforator flaps in the treatment of circular hot crush injuries in limbs with blood supply disorder.Methods:The study was a retrospective observational study. From April 2016 to December 2022, 4 cases with circular hot crush injuries in limbs with blood supply disorder were admitted to the Department of Burns and Plastic Surgery of the 988 th Hospital of Joint Logistics Support Force of PLA, including 3 males and 1 female, aged from 24 to 48 years. Among them, 2 cases were injured in the calf and 2 cases were injured in the forearm. After emergency debridement, the area of skin and soft tissue defects was from 20 cm×20 cm to 44 cm×20 cm. The patients had defects in tibialis anterior and posterior tibial arteries with a length of 13 to 18 cm, and in ulnar and radial arteries with a length of 9 to 12 cm. Flaps were designed and cut, including a flow-through anterolateral thigh perforator flap with area of 20 cm×9 cm to 24 cm×21 cm carrying the descending branch of the lateral circumflex femoral artery and the accompanying veins of 8 to 18 cm in length; and a flow-through posterior tibial artery perforator flap with area of 21 cm×13 cm and 20 cm×14 cm carrying the posterior tibial artery, the accompanying veins with a length of 14 and 17 cm respectively, and the great saphenous vein with a length of 22 and 21 cm. The circular hot crush injury wounds in the calf with blood supply disorder were repaired by a parallel combination of flow-through posterior tibial artery perforator flap and flow-through anterolateral thigh perforator flap, and the circular hot crush injury wounds in the forearm with blood supply disorder were repaired by a parallel combination of bilateral flow-through anterolateral thigh perforator flap, and the injured main vessels were reconstructed. The donor site wounds of flap were closed directly or treated with split-thickness skin grafts from abdomen. After surgery, the blood supply and survival of the flap and distal affected limb, the healing of wounds in the donor and recipient sites, the survival of the skin graft in the flap donor site were observed. During follow-up, the condition of flaps, the appearance, blood supply, and function of affected limbs were observed. At the last follow-up, the foot and ankle functions were evaluated according to the scoring standards of American Orthopedic Foot and Ankle Association, and the wrist and hand function was evaluated according to the trial standard of replantation of amputated upper limb function assessment of the Hand Surgery of Chinese Medical Association. Results:The flaps and distal affected limbs of 4 patients had good blood circulation and successfully survived after surgery. The wounds of 3 cases successfully healed, while one patient with circular hot crush injury in the forearm experienced exudation in the recipient site in the later stage, and the wound healed after re-expansion and suturing. The donor site wounds healed smoothly, and the skin grafts successfully survived. During follow-up of 12 to 24 months after surgery, the flaps were slightly swollen, the limbs had good appearance, normal blood circulation, and fine functional recovery. At the last follow-up, the foot and ankle function of 2 patients with circular hot crush injuries in the calf was evaluated as good in 1 case and commonly in 1 case; the wrist and hand function of 2 patients with circular hot crush injuries in the forearm was evaluated as excellent in 1 case and good in 1 case.Conclusions:The parallel combined flow-through perforator flap can reconstruct the damaged main blood vessels and repair the wound at the same time. It can not only effectively save the limb, but also restore part of the function of the affected limb. It is one of the effective methods to treat the circular hot crush injuries in limbs with blood supply disorder.

Objective: To investigate the value of transrectal ultrasonography(TRUS) in the preoperative diagnosis and tumor T-staging of anorectal malignant melanoma(ARMM). Methods: The clinical and ultrasound data of 19 patients(22 lesions) with ARMM confirmed by pathology from February 2008 to Apirl 2019 in Fujian Medical University Affiliated Union Hospital were collected. TRUS was performed within one week before pathological examination. The sonographic features were summarized, ultrasonic typing was performed according to the lesion size and growth pattern, and the accuracies of preoperative ultrasound diagnosis and tumor staging were analyzed. Results: Among the 22 lesions, 63.64%(14/22) were with a distance of ≤3 cm between the anal verge and the lower margin of tumor, 68.18%(15/22) with regular shape, 81.82%(18/22) with clear boundary, and 63.64%(14/22) with Adler blood flow grading from Ⅱ to Ⅲ. The preoperative TRUS diagnostic coincidence rate was 73.68%(14/19), and the tumor T-staging accuracy was 75%(12/16), respectively. Conclusions TRUS exhibits certain sonographic characteristics, with high preoperative diagnosis rate and tumor T-staging accuracy in anorectal malignant melanoma, which is of great guiding significance for clinical diagnosis and treatment.

Overexpression of adenosine triphosphate (ATP)-binding cassette subfamily G member 2 (ABCG2) in cancer cells is known to cause multidrug resistance (MDR), which severely limits the clinical efficacy of chemotherapy. Currently, there is no FDA-approved MDR modulator for clinical use. In this study, rociletinib (CO-1686), a mutant-selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), was found to significantly improve the efficacy of ABCG2 substrate chemotherapeutic agents in the transporter-overexpressing cancer cells and in MDR tumor xenografts in nude mice, without incurring additional toxicity. Mechanistic studies revealed that in ABCG2-overexpressing cancer cells, rociletinib inhibited ABCG2-mediated drug efflux and increased intracellular accumulation of ABCG2 probe substrates. Moreover, rociletinib, inhibited the ATPase activity, and competed with [I] iodoarylazidoprazosin (IAAP) photolabeling of ABCG2. However, ABCG2 expression at mRNA and protein levels was not altered in the ABCG2-overexpressing cells after treatment with rociletinib. In addition, rociletinib did not inhibit EGFR downstream signaling and phosphorylation of protein kinase B (AKT) and extracellular signal-regulated kinase (ERK). Our results collectively showed that rociletinib reversed ABCG2-mediated MDR by inhibiting ABCG2 efflux function, thus increasing the cellular accumulation of the transporter substrate anticancer drugs. The findings advocated the combination use of rociletinib and other chemotherapeutic drugs in cancer patients with ABCG2-overexpressing MDR tumors.

Objective: To explore the sonographic features of primary testicular lymphoma(PLT). Methods: The sonographic features of 16 PLT patients confirmed by surgery and pathology were retrospectively analyzed. Results: Sixteen PLT patients were typed into 3 ultrasonic patterns: ①The diffuse homogeneous echo type: 2 cases, the ultrasonic features were homogeneous sieve-like hypoechoic echo, the blood flow signal was rich and radially distributed, with interstisial fibrosis inapparently under-microscope. ②The diffuse uneven homogeneous echo type: 9 cases, the ultrasonic features lobulated or map-like heterogeneous hypoechoic features, the blood flow signal was rich in linear distribution, and interstisial fibrosis proliferated to form fibrous septa under-microscope. ③The focal nodular pattern echo type: 5 cases, the ultrasonic features were homogeneous hypoechoic nodules, the blood flow signal was few, interstisial fibrosis hyperplasia surrounding the tumor under-microscope. Conclusions Ultrasonic appearances of primary testicular lymphoma are closely related to its pathologic characteristics, the recognization of their relations contributes to improve the diagnostic value of ultrasound.

Overexpressing of ATP-binding cassette (ABC) transporters is the essential cause of multidrug resistance (MDR), which is a significant hurdle to the success of chemotherapy in many cancers. Therefore, inhibiting the activity of ABC transporters may be a logical approach to circumvent MDR. Olmutinib is an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), which has been approved in South Korea for advanced EGFR T790M-positive non-small cell lung cancer (NSCLC). Here, we found that olmutinib significantly increased the sensitivity of chemotherapy drug in ABCG2-overexpressing cells. Furthermore, olmutinib could also increase the retention of doxorubicin (DOX) and rhodamine 123 (Rho 123) in ABC transporter subfamily G member 2 (ABCG2)-overexpressing cells. In addition, olmutinib was found to stimulate ATPase activity and inhibit photolabeling of ABCG2 with [I]-iodoarylazidoprazosin (IAAP). However, olmutinib neither altered ABCG2 expression at protein and mRNA levels nor blocked EGFR, Her-2 downstream signaling of AKT and ERK. Importantly, olmutinib enhanced the efficacy of topotecan on the inhibition of S1-MI-80 cell xenograft growth. All the results suggest that olmutinib reverses ABCG2-mediated MDR by binding to ATP bind site of ABCG2 and increasing intracellular chemotherapeutic drug accumulation. Our findings encouraged to further clinical investigation on combination therapy of olmutinib with conventional chemotherapeutic drugs in ABCG2-overexpressing cancer patients.

Arsenic trioxide (ATO) is used as a chemotherapeutic agent for the treatment of acute promyelocytic leukemia. However, increasing drug resistance is reducing its efficacy. Therefore, a better understanding of ATO resistance mechanism is required. In this study, we established an ATO-resistant human epidermoid carcinoma cell line, KB/ATO, from its parental KB-3-1 cells. In addition to ATO, KB/ATO cells also exhibited cross-resistance to other anticancer drugs such as cisplatin, antimony potassium tartrate, and 6-mercaptopurine. The arsenic accumulation in KB/ATO cells was significantly lower than that in KB-3-1 cells. Further analysis indicated that neither application of P-glycoprotein inhibitor, breast cancer resistant protein (BCRP) inhibitor, or multidrug resistance protein 1 (MRP1) inhibitor could eliminate ATO resistance. We found that the expression level of ABCB6 was increased in KB/ATO cells. In conclusion, ABCB6 could be an important factor for ATO resistance in KB/ATO cells. The ABCB6 level may serve as a predictive biomarker for the effectiveness of ATO therapy.

Alectinib, an inhibitor of anaplastic lymphoma kinase (ALK), was approved by the Food and Drug Administration (FDA) for the treatment of patients with ALK-positive non-small cell lung cancer (NSCLC). Here we investigated the reversal effect of alectinib on multidrug resistance (MDR) induced by ATP-binding cassette (ABC) transporters, which is the primary cause of chemotherapy failure. We provide the first evidence that alectinib increases the sensitivity of ABCB1- and ABCG2-overexpressing cells to chemotherapeutic agents in vitro and in vivo. Mechanistically, alectinib increased the intracellular accumulation of ABCB1/ABCG2 substrates such as doxorubicin (DOX) and Rhodamine 123 (Rho 123) by inhibiting the efflux function of the transporters in ABCB1- or ABCG2-overexpressing cells but not in their parental sensitive cells. Furthermore, alectinib stimulated ATPase activity and competed with substrates of ABCB1 or ABCG2 and competed with [125I] iodoarylazidoprazosin (IAAP) photolabeling bound to ABCB1 or ABCG2 but neither altered the expression and localization of ABCB1 or ABCG2 nor the phosphorylation levels of AKT and ERK. Alectinib also enhanced the cytotoxicity of DOX and the intracellular accumulation of Rho 123 in ABCB1-overexpressing primary leukemia cells. These findings suggest that alectinib combined with traditional chemotherapy may be beneficial to patients with ABCB1- or ABCG2-mediated MDR.
Adenosine Triphosphatases ; Carcinoma, Non-Small-Cell Lung ; Doxorubicin ; Drug Resistance, Multiple* ; Drug Therapy ; Humans ; In Vitro Techniques* ; Leukemia ; Lymphoma ; Parents ; Phosphorylation ; Phosphotransferases ; Rhodamine 123 ; United States Food and Drug Administration

Purpose To explore ultrasonic characteristics of complex fibroadenoma (CF) of breast and its differential diagnosis from invasive ductal carcinoma (IDC).Materials and Methods Sixty-one patients with CF and 81 patients with IDC from January 2010 to February 2016 in the Affiliated Union Hospital of Fujian Medical University were enrolled,and the CF and IDC were confirmed by pathology after operation.The preoperative sonographic findings of CF and IDC were retrospectively analyzed and compared.Results Fifty-nine cases with CF were detected by ultrasonography in 61 cases.Ultrasonic findings of 59 cases of CF showed as follows:79.7％ (47/59) of the CF cases with an aspect ratio ＜0.7;79.7％ (47/59) with blood flow signal of 0 to 1 level;72.9％ (43/59) with liquid dark area,including 67.8％ (40/59) with round or oval cystic fluid area showing local sievelike or diffused distribution;72.9％ (43/59) with irregular shape;61.0％ (36/59) with obscure boundary;61.0％ (36/59) with the largest diameter of tumor ＜2 cm;59.3％ (35/59) with calcifications,including 33.9％ (20/59) with coarse calcification,25.4％ (15/59) with microcalcification and 85.7％ (30/35) with distribution of calcified lesions along the linear high echo zone;54.2％ (32/59) with echo heterogenicity showing linear high echo and high echo in the middle and low echo.There were significant differences in the detection rates of internal regular fluid area,internal linear high echo or high echo area,and alcification distribution along the linear high echo between CF and IDC lesions (all P＜0.01).Compared with IDC,there were higher rates in CF for detecting lesions of largest diameter of tumor ＜2 cm,more regular shape,more clear border,poorer blood flow signal (P＜0.05),but there were lower rates in CF for detecting axillary lymph node enlargement,lesions with edge or spiculation and hyperechoic halo (P＜0.01).Conclusion Color Doppler ultrasonography is of great value in the diagnosis of CF and its differential diagnosis from IDC.

Objective To evaluate the one-year clinical outcomes in patients with the vulnerable plaque sealing with drug-eluting stents for the treatment of intermediate coronary stenosis. Methods 327 patients with an-giographically intermediate lesions(diameter stenosis 50%~70%)with the vulnerable plaque which were detected by 64 slice coronary CT were prospectively enrolled. Patients were divided into medical therapy group (n = 160) and sirolimus-eluting stent group group(n=160). The incidences of one-year major adverse cardiovascular events (MACE)was evaluated(cardiac death,myocardial infarction ,revascularization). Results The MACE tended to be lower in the sirolimus-eluting stent group than medical therapy group(3.13%vs. 10%,log-rankχ2=6.62,P=0.01). The incident of cardiac death and myocardial infarction were lower in the sirolimus-eluting stent group than medical therapy group(1.25%vs. 5.63%,log-rankχ2=4.61,P=0.03). Conclusion The treatment of the siroli-mus-eluting stent can reduce MACE for the paitents with the vulnerable plaque of intermediate coronary stenosis than medical therapy only.