AIM:To construct and breed infertility model mice with ATP/GTP binding carboxypeptidase 1(Agtpbp1)gene knockout homozygote(Agtpbp1-/-)by CRISPR/Cas9 technology,so as to provide an animal model for the subsequent exploration of the pathogenesis of Agtpbp1 gene in male sterility.METHODS:The CRISPR/Cas9 technology was used to obtain Agtpbp1 gene knockout heterozygote(Agtpbp1+/-)mice according to the data of the main protein functional region of Agtpbp1 gene and combined with Cas9 nuclease.The obtained Agtpbp1+/-mice were mated,and their offspring were genotyped by PCR and agarose gel electrophoresis.The expression of Agtpbp1 at different levels was detected by RT-PCR,Western blot and immunohistochemical staining to support the identification results.The HE staining was used to observe the mouse cerebellum and eyeball structure to analyze the effect of Agtpbp1 gene knockout on Purkinje cells and photoreceptor cells.The symptoms of ataxia in mice were observed in combination with behavioral tests.The growth of mice was observed,and the changes of testicular tissue volume and weight of male mice were analyzed.The HE staining was used to observe the changes of testicular structure,and PAS staining was used to observe the changes of testicular germ cell cycle.Finally,sperm analyzer was used to analyze the sperm motility,so as to analyze the growth and develop-ment of the mice.RESULTS:The male and female Agtpbp1+/-mice could continue to mate,and three genotypes,Agtpbp1 wild-type(Agtpbp1+/+),Agtpbp1+/-and Agtpbp1-/-,were obtained.The genotypes of the offspring mice were successfully identified by PCR.The results of RT-PCR,Western blot and immunohistochemical staining verified the successful con-struction of Agtpbp1-/-mouse model at different levels(P<0.05).The results of HE staining showed that Purkinje cells were lost in the cerebellum of Agtpbp1-/-mice and the number of photoreceptor cells in the eyeball was reduced.Behavioral tests confirmed that Agtpbp1-/-mice had ataxia symptoms such as motor dysfunction and uncoordinated movements.Com-pared with control group,the testicular volume and weight of Agtpbp1-/-mice were significantly reduced.The results of HE staining showed a very small amount of sperm in the testis of Agtpbp1-/-mice.Combined with the sperm analyzer,it was ob-served that the sperm motility,vitality and movement rate of Agtpbp1-/-mice were significantly lower than those of the con-trol mice.Testicular sections with PAS staining showed cell cycle arrest of the sperm from Agtpbp1-/-mice.CONCLU-SION:In this study,Agtpbp1 knockout mice were successfully bred.The deletion of Agtpbp1 caused the arrest of sper-matogenic cell differentiation and the decrease in sperm motility in adult male mice,resulting in infertility.At the same time,it provides a new experimental tool for further exploring the molecular mechanism of Agtpbp1-induced male sterility.

AIM:To construct and breed infertility model mice with ATP/GTP binding carboxypeptidase 1(Agtpbp1)gene knockout homozygote(Agtpbp1-/-)by CRISPR/Cas9 technology,so as to provide an animal model for the subsequent exploration of the pathogenesis of Agtpbp1 gene in male sterility.METHODS:The CRISPR/Cas9 technology was used to obtain Agtpbp1 gene knockout heterozygote(Agtpbp1+/-)mice according to the data of the main protein functional region of Agtpbp1 gene and combined with Cas9 nuclease.The obtained Agtpbp1+/-mice were mated,and their offspring were genotyped by PCR and agarose gel electrophoresis.The expression of Agtpbp1 at different levels was detected by RT-PCR,Western blot and immunohistochemical staining to support the identification results.The HE staining was used to observe the mouse cerebellum and eyeball structure to analyze the effect of Agtpbp1 gene knockout on Purkinje cells and photoreceptor cells.The symptoms of ataxia in mice were observed in combination with behavioral tests.The growth of mice was observed,and the changes of testicular tissue volume and weight of male mice were analyzed.The HE staining was used to observe the changes of testicular structure,and PAS staining was used to observe the changes of testicular germ cell cycle.Finally,sperm analyzer was used to analyze the sperm motility,so as to analyze the growth and develop-ment of the mice.RESULTS:The male and female Agtpbp1+/-mice could continue to mate,and three genotypes,Agtpbp1 wild-type(Agtpbp1+/+),Agtpbp1+/-and Agtpbp1-/-,were obtained.The genotypes of the offspring mice were successfully identified by PCR.The results of RT-PCR,Western blot and immunohistochemical staining verified the successful con-struction of Agtpbp1-/-mouse model at different levels(P<0.05).The results of HE staining showed that Purkinje cells were lost in the cerebellum of Agtpbp1-/-mice and the number of photoreceptor cells in the eyeball was reduced.Behavioral tests confirmed that Agtpbp1-/-mice had ataxia symptoms such as motor dysfunction and uncoordinated movements.Com-pared with control group,the testicular volume and weight of Agtpbp1-/-mice were significantly reduced.The results of HE staining showed a very small amount of sperm in the testis of Agtpbp1-/-mice.Combined with the sperm analyzer,it was ob-served that the sperm motility,vitality and movement rate of Agtpbp1-/-mice were significantly lower than those of the con-trol mice.Testicular sections with PAS staining showed cell cycle arrest of the sperm from Agtpbp1-/-mice.CONCLU-SION:In this study,Agtpbp1 knockout mice were successfully bred.The deletion of Agtpbp1 caused the arrest of sper-matogenic cell differentiation and the decrease in sperm motility in adult male mice,resulting in infertility.At the same time,it provides a new experimental tool for further exploring the molecular mechanism of Agtpbp1-induced male sterility.

Objective To explore the automated identification and diameter measurement methods for inferior vena cava (IVC) based on clinical ultrasound images of IVC. Methods An automated identification and localization method based on topology and automatic tracking algorithm was proposed. Tracking algorithm was used for identifying and continuously locating to improve the efficiency and accuracy of measurement. Tests were conducted on 18 sets of ultrasound data collected from 18 patients in intensive care unit (ICU),with clinicians' measurements as the gold standard. Results The recognition accuracy of the automated method was 94.44％ (17/18),and the measurement error of IVC diameter was within the range of±1.96s (s was the standard deviation). The automated method could replace the manual method. Conclusion The proposed IVC automated identification and localization algorithm based on topology and automatic tracking algorithm has high recognition success rate and IVC diameter measurement accuracy. It can assist clinicians in identifying and locating IVC,so as to improve the accuracy of IVC measurement.

Objective To summarize the clinical treatment experience of carbapenem-resistant Klebsiella pneumoniae (CRKP) infection after renal transplantation in donation after cardiac death (DCD) era. Methods Clinical data of 10 donors and 17 recipients with CRKP infection after DCD renal transplantation from January 2015 to January 2019 were retrospectively analyzed. Both donors and recipients received bacterial culture and drug sensitivity test. Clinical manifestations, treatment and outcome of CRKP-infected recipients were recorded. Results Seven donors were infected with CRKP. After pretreatment, CRKP in 2 cases turned negative, CRKP in 5 donors did not turn negative. All renal grafts were treated with tigecycline+meropenem+voriconazole lavage to prevent infection. Among 17 recipients with CRKP infection, 11 cases were positive for blood culture, 10 positive for urine culture, 3 positive for sputum culture, 3 positive for incisional secretion and 3 positive for retroperitoneal drainage. Clinical manifestations included fever in 8 cases, rupture and hemorrhage of the transplant renal artery in 7 cases or thrombosis in the transplant renal artery in 1 case, bladder irritation sign in 3 cases and cough with brick red jelly-like sputum in 1 case, respectively. Five patients were treated with tigecycline+meropenem, 1 patient suffered from renal graft loss and 4 recipients died. Twelve patients were treated with ceftazidime-avibactam +meropenem, 3 patients presented with renal graft loss and 1 recipient died. Conclusions CRKP-infected donor is not the absolute contraindication of renal transplantation. Pretreatment of donor infection and early administration of sufficient sensitive antibiotics can cure CRKP infection and improve the clinical prognosis of renal transplant recipients.

Objective:To explore the risk factors of pulmonary infection in elderly (aged 60+ years) kidney transplant recipients.Methods:The clinical data were retrospectively analyzed for 119 elderly kidney transplant recipients from January 2010 to January 2019 . According to whether or not pulmonary infection occurred after renal transplantation, the recipients were divided into infected group (n=40) and non-infected group (n=79). Clinical data was analyzed for two groups. The relevant risk factors of gender, age, donor type, body mass index, history of smoking, preoperative dialytic time, preoperative dialysis, immune induction, immune maintenance, presence or absence of delayed graft function, leucopenia, serum creatinine before infection, venous hormone shock therapy or not, diabetic history before or after surgery, history of coronary heart disease, history of hepatitis B virus, prophylactic dosing of compound sulfamethoxazole, prophylactic valganciclovir or ganciclovir, were examined by univariate analysis and multivariate logistic regression analysis.Results:The incidence of pulmonary infection in elderly kidney transplant recipients was 33.6% (40/119). In infected group, 15 patients died of severe pulmonary infection with a mortality rate of 37.5%(15/40). History of smoking (OR=10.58, 95%CI: 1.98-56.40, P=0.006), venous hormone shock therapy (OR=25.06, 95%CI: 4.25-147.71, P<0.001) and preoperative dialytic time (OR=1.032, 95%CI: 1.003-1.062, P=0.033) were the risk factors for pulmonary infection in elderly kidney transplant recipients. Conclusions:The incidence and mortality of lung infection are higher in elderly kidney transplant recipients. Smoking history, venous hormone shock therapy and long preoperative dialytic are associated with pulmonary infection in elderly kidney transplant recipients.

Wolfiporia cocos (F. A. Wolf) has been praised as a food delicacy and medicine for cen-turies in China. Here, we present the genome and transcriptome of the Chinese strain CGMCC5.78 of W. cocos. High-confidence functional prediction was made for 9277 genes among the 10,908 total predicted gene models in the W. cocos genome. Up to 2838 differentially expressed genes (DEGs) were identified to be related to sclerotial development by comparing the transcriptomes of mycelial and sclerotial tissues. These DEGs are involved in mating processes, differentiation of fruiting body tissues, and metabolic pathways. A number of genes encoding enzymes and regulatory factors related to polysaccharide and triterpenoid production were strikingly regulated. A potential triter-penoid gene cluster including the signature lanosterol synthase (LSS) gene and its modified compo-nents were annotated. In addition, five nonribosomal peptide synthase (NRPS)-like gene clusters, eight polyketide synthase (PKS) gene clusters, and 15 terpene gene clusters were discovered in the genome. The differential expression of the velevt family proteins, transcription factors,carbohydrate-active enzymes, and signaling components indicated their essential roles in the regu-lation of fungal development and secondary metabolism in W. cocos. These genomic and transcrip-tomic resources will be valuable for further investigations of the molecular mechanisms controlling sclerotial formation and for its improved medicinal applications.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Objective To avoid the adverse consequences caused by human intervention in the process of blood transfusion and to promote the improvement of the quality index of blood transfusion specialist by the construction and application of closed loop management system for blood transfusion in mobile nursing information system. Methods In June 2014, the Nursing Department, the Information Department, the Blood Transfusion Department cooperated to analyzed the a variety of existing hidden dangers in blood transfusion process. After reengineering the related process of blood transfusion and setting up a certain of warning reminders, the closed loop management system for blood transfusion was finally constructed with the help of the research and development company using the mobile information system. A total of 2 202 cases from March to May 2014 before the application of the closed loop management system for blood transfusion in mobile nursing information system and 2 916 cases from January to March 2015 after the application of the system were investigated and analyzed, so as to compare the quality index and the incidence of adverse medical events. Results The implementation rate of blood transfusion in 4 hours, the abnormal cases in inspection per hour, the implementation rate of the inspection 15 min before transfusion, the cases of non-replacement of transfusion device in 4 hours all improved after the application of the closed loop management system for blood transfusion in mobile nursing information system, and the differences were statistically significant (P<0.01). In addition, there was no case of adverse medical event after the application of the closed loop management system for blood transfusion in mobile nursing information system. Conclusions Transforming the blood transfusion nursing process through information technology can realize the real-time monitoring of transfusion quality indicators and reduce the occurrence of adverse medical events, so as to ensure the transfusion safety.

Objective To analyze and summarize clinical characteristics and treatment of diffuse alveolar hemorrhage syndrome (DAHS)complication after renal transplantation.Methods Clinical data of one patient,admitted to the 309 th Hospital of People's Liberation Army in December 201 2, who was complicated with DAHS after renal transplantation,were obtained.The incidence,diagnosis and treatment courses of DAHS were retrospectively analyzed.Literature review was conducted to summarize clinical experience.Results The patient was clinically manifested with respiratory failure,progressive aggravation of hemoptysis and anemia.Imaging examination revealed that diffusive infiltration of bilateral lungs was aggravated.After the diagnosis of DAHS was confirmed,adrenal cortical hormone (hormone)shock and anti-infectious medication therapies were timely delivered to actively prevent and treat relevant complications.The patient was successfully healed.Until the submission date,the patient presented with normal renal function and no pulmonary complications were noted.Conclusions DAHS is a rare and fatal complication after renal transplantation.Early diagnosis, active anti-infection therapy and timely administration of large-dose hormone shock treatment determine the success of clinical treatment.

Objective To analyze the clinical efficacy of multiple renal arteries on outcomes of renal donors and recipients in hand-assisted retroperitoneoscopic donor nephrectomy.Method From 2012 to 2014,121 patients underwent hand-assisted laparoscopic donor nephrectomy,including 92 cases of a single renal artery and 29 cases of multiple arteries.Donor and recipient outcomes for single artery and multiple arteries allografts were compared.Result The study included 121 pairs of donors and recipients.The demographic characteristics between multiple renal artery group and single renal artery group had no significant difference.The operative time,blood loss,postoperative complications,and hospital stay had no significant difference between two groups.Cold ischemia time and warm ischemia time in multiple renal artery group were longer than single donor renal artery group (128.5 ± 13.2 vs.50.2 ± 17.3 min,P＜0.001;196.0 ± 63.3 vs.154.1 ± 55.2 min,P=0.002,respectively).The operative time in multiple renal artery group was longer than in single renal artery group (213.5 ± 28.2 vs.182.2 ± 31.1 min,P＜0.001).There was no significant difference in blood loss,vascular complications and ureternal complications between two groups.The renal functions of two groups were likewise within one year.Conclusion There was no statistically significant difference in clinical efficacy between hand-assis-ted retroperitoneoscopic donor nephrectomy with multiple renal arteries and single artery.The use of these grafts was safe for both recipients and donors.