Objective:To prepare and screen monoclonal antibodies against Herpes simplex virus-1(HSV-1),and develop a double antibody sandwich quantitative enzyme-linked immunosorbent assay( Q-ELISA) for detection of HSV-1 particle. This method was used to control the quality of viral particle in the developing and manufacturing process of HSV-1. Methods: BALB/c mice was immunized with HSV-1 to prepare monoclonal antibodies. A double antibody sandwich Q-ELISA was developed to determine concentration of HSV-1 particle,which was based on the neutralizing monoclonal antibody 1F6 as capture antibody,and 2B1 as HRP-conjugated antibody. The performance of the reagent was evaluated,including specificity,sensitivity,precision,accuracy and linear. And the relation between the amount of virus detected by this method and the virus titer was analyzed by regression analysis method. Results: The Q-ELISA for HSV-1 particle was developed. The quantitation scope was 0. 125-2 μg/ml, the coefficient correlation was 0. 995 5, the limit of detection was 0. 125 μg/ml, the recovery was between 85. 6% and 107. 1%, the variation coefficient was lower than 10%, and the reagent does not react with other samples except HSV-1 antigen. This method has a good correlation with virus titer. Conclusion:The Q-ELISA for HSV-1 particle was successfully developed,which provide a new approach for rapid and quantitative detection of HSV-1 antigen.

Objective:To investigate the clinical effect of free deep femoral artery third perforating flap repaired soft tissue loss after Pilon fracture surgery in I stage.Methods:Fifteen patients were treated from April, 2013 to January, 2020. Miller AO classification: 8 cases 43-C1, 4 cases 43-C2 and 3 cases 43-C3. All cases were accompanied with severe soft tissue contusion and skin necrosis. After fracture reduction, soft tissue defects, internal fixation exposure and tendon exposure around the wound. Free deep femoral artery third perforating flap (3.5 cm ×15.5 cm to 5.5 cm×12.5 cm) for the repair of soft tissue defects around ankle in the I stage, the blood vessels of the flap were end-to-side anastomosed with vessels of the posterior tibial or anterior tibial. Regular follow-up after surgery.Results:One case of venous crisis occurred, other 14 cases survived, were followed-up from 5 to 18 months, the ankle joint function was good, did not affect the foot shoes, with excellent color and texture, the flap restored protective sensation, and leaving only linear scar, no muscle adhesion.Conclusion:Free deep femoral artery third perforating flap repaired soft tissue loss of surgical incision after fracture operated than significantly reduce the postoperative fracture infection and protect the blood supply around the fracture. It is an effective method of repair.

OBJECTIVE: To study the correlation between dosage and curative effect of blood coagulation factor VIII in the prevention and treatment of haemophilia A in children and to determine the suitable dose for prevention of hemophilia in developing countries. METHODS: For different body weights of child patient, every time we used the same dosage of blood coagulation factor VIII (250 U each time, 3 times a week) and observed and recorded the number of hemorrhages in child patients. Then we compared the number of hemorrhages with children without treatment to determine the curative effect. According to the different body weights, we calculated the dosage of VIII factor of blood coagulation per kilogram (hereinafter referred to as the dose), and used Spearman correlation coefficient to study the correlation between dose and curative effect. RESULTS: The number of hemorrhages in 58 child patients before the treatment was 4.36 ± 1.78, while after the treatment was 2.22 ± 1.04 (t=7.91, P<0.001). The Spearman correlation coefficient of child patients of 5-10 U/kg was -0.421 (P=0.005); the Spearman correlation coefficient of child patients of 10-15 U/kg was -0.331 (P=0.030); the Spearman correlation coefficient of child patients over 15 U/kg was -0.16 (P=0.325). CONCLUSION Prevention and treatment can significantly reduce the times of hemorrhage in hemophilia patients.
Blood Coagulation ; Child ; Factor VIII ; administration & dosage ; therapeutic use ; Hemophilia A ; therapy ; Hemorrhage ; prevention & control ; Humans

Objective: To investigate the clinical value of modified transperineal template-guided prostate biopsy (mTTPB) in the detection of prostate cancer. Methods: A total of 217 patients were enrolled in this study. All the patients were randomly divided into 2 groups. The control group (n=112) underwent transperineal template-guided prostate biopsy (TTPB) which was traditional transperineal template-guided 11-region biopsy. On the basis of the control group, the apex of prostate was divided into four areas for biopsy in the observation group (mTTPB). The positive rate of apex and the incidence of complications were analyzed. The prostatic specimens from the radical prostatectomy underwent whole mount sections examination. The prostate biopsy results were compared with the postoperative pathological results. Results: The average age of the control group and the observation group were (68.5±7.9) years and (67.3±8.5) years, PSA were (31.2±18.9) ng/ml and (29.7±19.5) ng/ml, prostate volume were (44.6±15.2) ml and (41.3±17.3) ml, respectively. In the control group, the positive rates of prostate cancer in 1-10 region were 24.1% (27/112), 27.7%(31/112), 23.2% (26/112), 28.6% (32/112), 26.8% (30/112), 25.0% (28/112), 26.8% (30/112), 19.6% (22/112), 25.9% (29/112), 25.0% (28/112), respectively, with an average of 25.3%. In the observation group, the positive rates in 1-10 region were 27.6% (29/105), 28.6% (30/105), 22.9% (24/105), 26.7% (28/105), 25.7% (27/105), 24.8% (26/105), 27.6% (29/105), 21.9% (23/105), 27.6% (29/105), 26.7% (28/105), respectively, with an average of 26.0%. There was no statistical difference between the two groups (P=0.904). The positive rate of apical prostate cancer in the control group and observation group was 37.5% (42/112) and 44.8% (47/105), respectively, and there was no statistical difference between the two groups (P=0.277). Patients were grouped according to PSA>20 ng/ml and PSA≤20 ng/ml. When PSA>20 ng/ml, the positive rate of apex was 58.6% (34/58) and 56.6% (30/53) respectively in the control group and the observation group, and there was no statistical difference between the two groups (P=0.830). When PSA≤20 ng/ml, the positive rate of apex was 14.8% (8/54) in the control group and 32.7% (17/52) in the observation group, with statistically significant differences (P=0.030). Before radical prostatectomy, 12 cases (57.1%) in the control group and 19 cases (73.1%) in the observation group showed apical invasion by biopsy. Results of whole mount sections examination in the control group showed that there were 19 cases (90.5%) with apical invasion, which was statistically different from that before surgery (P=0.035). The results of whole mount sections examination in the observation group showed that there were 23 cases (88.5%) with apex invasion, which had no statistical difference compared with that before surgery (P=0.291). There were no significant differences in the incidence of hematuria, fever, urinary retention and perineal discomfort between the observation group and the control group (all P>0.05). Conclusions mTTPB can significantly improve the detection rate of apical prostate cancer without increasing the incidence of complications, especially for patients with PSA≤20 ng/ml. Hence is safe and efficacious.

Objective:To analyze the risk of missed diagnosis in patients with PI-RADS score>3 and negative prostate initial biopsy and to explore its risk factors.Methods:The clinical data of 268 patients with negative prostate biopsy in Northern Jiangsu People's Hospital from May 2013 to December 2018 were retrospectively analyzed. The patients were divided into observation group (PI-RADS score>3) and control group (PI-RADS score≤ 3) according to different PI-RADS scores. There were insignificant differences in age [(67.4(60.0, 74.0)years and 65.6(66.5, 72.0)years], prostate volume of initial biopsy [62.4(40.0, 72.0)ml and 60.8(38.0, 77.0)ml], biopsy cores [ 20.6(18.0, 22.0)cores and 20.4(18.0, 22.0)cores] between the observation group (n=124) and the control group(n=144)(all P>0.05). But there were significant differences in PSA [17.5(6.5, 23.0)ng/ml and 11.5(6.3, 12.0)ng/ml], PSAD[0.316(0.128, 0.363)ng/ml 2 and 0.211(0.106, 0.256)ng/ml 2], prostate inflammation of the initial biopsy [70 (56.5%) and 32 (22.2%)] between the observation group and the control group(all P<0.05). According to the follow-up results after the initial biopsy, the two groups of repeated biopsy were compared.Furthermore, Logistic regression was used to conduct univariate and multivariate analysis to explore the risk factors of patients with PI-RADS>3 for positive repeated biopsy. At the same time, the receiver operating characteristic curve (ROC curve) was used to analyze the accuracy of the risk factors. Results:There were significant differences in repeated biopsy rate [ 27.4%(34/124)and 14.6%(21/144)], CsPCa detection rate[ 41.4%(14/34) and 4.8%(1/21)]between the observation group and the control group(all P<0.05). The positive rate of repeated biopsy in the observation group (41.1%) was higher than that in the control group (23.8%), but there was no statistical difference ( P=0.248). The risk of positive repeated biopsies in the observation group was 2.24 times than that in the control group. Univariate analysis found repeated biopsy PSA ( P =0.02, OR=1.438, 95% CI 1.161-1.896), PSA ratio (repeated biopsy PSA/initial biopsy PSA) ( P=0.011, OR=10.087, 95% CI 1.714-59.36) were risk factors for positive of repeated biopsy in patients with PI-RADS score >3. Multivariate analysis also found that repeated biopsy PSA ( P=0.017, OR=1.15, 95% CI 1.076-2.123), PSA ratio ( P=0.032, OR=10.2, 95% CI 0.883-116.168) were risk factors for positive repeated biopsy. ROC curve analysis, the accuracy of repeated biopsy PSA (AUC=0.971, P<0.001, 95% CI 0.926-1.000), PSA ratio (AUC=0.839, P=0.001, 95% CI0.707-0.971) to predict positive of repeated biopsy were high. The cut-off values were 21.3 ng/ml and 1.4, respectively. The accuracy was higher when combines repeated biopsy PSA with PSA ratio (AUC=0.993, P<0.001, 95% CI 0.974-1.000). Conclusions:Patients with negative PI-RADS score > 3 have a higher risk of missed diagnosis of CsPCa than those with PI-RADS score≤3. When PSA>21.3 ng/ml and PSA ratio>1.4 during follow-up, the possibility of missed diagnosis in the initial biopsy is high.

Functional genetic screening as an important method for exploring biological processes, diseases development research and functional annotation of genetic elements, has been widely used in pharmaceutical research, new therapeutic targets identifying and screening, and tumor resistance. CRISPR-Cas9 (Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9) is the newest tool in the geneticist's toolbox, allowing researchers to edit genome with unprecedented ease, accuracy and high-throughput. CRISPR-Cas9 system provides a high-throughput, practical and efficient tool for the discovery of functionally important genes responsible for certain phenotypes. In this review, we summarize the characterization of CRISPR/Cas9 system and applications of this new genetic toolkit in functional genetic screening.

The present study retrospectively analyzed the clinical data of 137 patients who underwent prostate in North Jiangsu People's Hospital from June 2020 to May 2021. All patients underwent peripheral prostatic nerve block anesthesia (PPNB). The observation group received 1% ropivacaine 32 ml local, and the control group received the same dose of lidocaine. There was no significant difference in general data before puncture between the two groups ( P>0.05). All 137 cases were performed by the same surgeon. The number of puncture needles in the observation group and the control group was (20.2±2.8) and (20.2±2.9), respectively, and the difference was not statistically significant ( P>0.05). The visual analogue scores (VAS-1) of pain during puncture in the observation group and the control group were (2.62±0.74) and (2.48±0.79) points, respectively. The visual numeric score (VNS-1) was (3.03±0.88) points and (3.15±0.80) points, respectively, and there was no significant difference ( P>0.05). 30 min after puncture, VAS-2 was (0.48±0.53) points and (0.30±0.47) points, VNS-2 was (3.31±0.48) points and (3.55±0.71) points, respectively.The differences were statistically significant ( P<0.05). There was no significant difference in overall complication rate between the two groups ( P=0.661).

Objective: To analyze the clinical characteristics of children with neuroblastoma (NB) complica-ted with lung or pleural metastasis, further to explore the correlation between characteristics and short-term outcome of NB, so as to provide a basis for clinical diagnosis and treatment. Methods: A retrospective analysis was performed concerning the age of onset, clinical features, treatment and outcome of 36 patients with NB who were admitted at Blood Tumor Center, Beijing Children′s Hospital of Capital Medical University from December 2007 to December 2017.The diagnostic criteria, therapeutic regimen and therapeutic efficacy criteria of the enrolled children were all based on the NB protocol of Beijing Children′s Hospital of Capital Medical University (BCH-NB-2007), the clinical stage was based on international clinical stage of neuroblastoma (INSS stage), and stratified treatment was conducted according to the BCH-NB risk grouping standard.The follow-up period lasted till October 31, 2018. Results: (1)The common clinical features of grouped children: 36 patients were selected into the group, accounted for 5.99% (36/601 cases) in total hospitalized NB children, they were less than 10 years old, 10 cases under 18 months, and the median age was 29.5 months (9-105 months); 20 cases were male, and 16 cases were female; the primary tumor was located in the retroperitoneal site in 19 cases, accounting for 52.78%, 9 cases in adrenal site, accounting for 25.00%, and 8 cases in mediastinal site, accounting for 22.22%.Risk groups: 29 cases were in high-risk group, 6 cases were in medium-risk group and 1 case was in low-risk group.The main symptoms were of pain onset in 8 cases, fever in 6 cases, local mass in 6 cases, abdominal mass in 4 cases, mediastinal mass in 3 cases, paleness in 3 cases, subcutaneous nodules in 2 cases, abdominal distension in 2 cases, lower limb swelling in 1 case, and diarrhea in 1 case. Among them, 16 cases had respiratory system symptoms first, accounting for 44.4%.(2)Laboratory examination: there were 35 patients of neuron-specific enolase (NSE) ≥25 μg/L on the initial diagnosis, of which 11 cases were more than 370 μg/L, the value of serum lactate dehydrogenase (LDH) ≥717.5 U/L in 25 patients, accounting for 69.44%, and 10 cases were accompanied by N-myc gene amplification.(3)Imaging examination: on the first diagnosis, chest/abdomen CT showed 75.75%(24/33 cases) of pleural or lung involvement, PET-CT showed 81.8% (27/33 cases) of pleural or lung involvement, B-ultrasound showed 41.67%(5/12 cases) of pleural or lung involvement; 3 imaging examinations showed: 1 positive in 16 cases, 2 positive in 16 cases, and 3 positive in 4 cases.(4)Comparison of clinical features of pleural and pulmonary involvement: among the 30 children with pleural involvement, 6 cases had respiratory symptoms, 3 cases had respiratory symptoms on the 6 cases with pulmonary involvement only, and 4 cases with pleural and pulmonary involvement had no respiratory symptoms at first diagnosis.(5)Treatment and outcome: 2 cases died because of critical condition after diagnosis, 5 cases didn′t receive the law treatment, 29 cases accepted law stratification treatment, among them, 1 case in low risk group, INSS-Ⅳ, alleviate current (CR); 4 cases in the moderate group, INSS-Ⅳ, CR in 2 cases, partial response (PR) in 1 case, progress in 1 case, new tumor foci occurring after chemotherapy discontinuation for 13 months. Twenty-four cases were in the high-risk group, event occurred in 7 cases (29.17%) of them, 1 case had progression by postoperative evaluation, 4 cases of progression at 1.5, 2.0, 3.0 and 6.0 months after cessation of chemotherapy, and 2 cases had recurrence at 11 and 17 months after cessation of chemotherapy.Overall survival rate (OS) was 41.4% for all children analyzed by Kaplan-Merier, and 32.9% of them were predicted to have 3-year event-free survival. Conclusions Children with pulmonary or pleural metastasis of neuroblastoma have no specific respiratory symptoms.CT scan might be a useful method for diagnosing the group Ⅳ children with pulmonary or pleural metastasis of neuroblastoma.Moreover, there seemed to be no significant correlation between the N-myc gene expression and survival prognosis of these children.