Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.

Objective:To observe the effect of Linggui Zhugan Tang on gut microbiota in ob/ob mice, in order to explore whether its bone protective effect is related to its regulatory effect of gut microbiota. Method:The ob/ob mice were divided into control group, model group, Linggui Zhugan Tang group, and Atorvastatin group. After administration for 4 weeks, morphological changes in intestine and bone were observed by htoxylin eosin(HE) staining. The protein expression of alkaline phosphatase(ALP) was detected by Western blot in bone. The expression of farniol X receptor(FXR) was detected by in immunofluorescence staining. And the content of interleukin-6(IL-6) in serum was analyzed by the enzyme-linked immunosorbent assay(ELISA) method. Gut microbiota was analyzed by 16S rRNA. Result:Compared with control group, the trabecular bone of the femur became thinner, the bone marrow cavity was infiltrated with fat, and the epithelium of the small intestine was damaged in the model group. The protein expressions of ALP and FXR decreased, while the content of IL-6 in serum was increased in the model group. Compared with model group, the protein expressions of ALP and FXR increased, while the content of IL-6 in serum was decreased in the Linggui Zhugan Tang group. At the genus level, compared with control group, the abundance of Lachnospiraceae_UCG-001 increased (P<0.05), while that of norank_f__Bacteroidales_S24-7_group decreased in model group (P<0.01). Lachnospiraceae_UCG-001 was positively correlated with the weight of body (r=0.661), while norank_f__Bacteroidales_S24-7_group was negatively correlated with the weight of body (r=-0.622). Conclusion:Linggui Zhugan Tang has a protective effect on bone injury in ob/ob mice, and its mechanism may be related to its regulation of gut microbiota.

OBJECTIVE: To review the development of adult and pediatric liver transplantation in Tianjin First Center Hospital, and to enhance academic exchanges, improve technological innovation, and jointly promote the progress and maturity in the field of liver transplantation. METHODS: The development of liver transplantation in Tianjin First Center Hospital was analyzed. The clinical data of adult and pediatric liver transplantation from September 1998 to September 2018 were collected. The important events and technological innovation achievements of liver transplantation during the 20 years were summarized. RESULTS: The first clinical liver transplantation was attempted in Tianjin First Central Hospital in April 1980. The first long-term survival adult liver transplantation in China was completed in 1994 (11 years survival after the operation). The specialized team of liver transplantation was formally established in September 1998. The 20-year clinical exploration and progress reflected the characteristics of era changes and technological innovation during the rapid development of liver transplantation in China. Our center performed liver re-transplantation in January 1999, reduced-size pediatric liver transplantation in August 2000. In May 2001, we organized the formulation for the preventive and treatment plan for hepatitis B recurrence after liver transplantation. We performed combined liver and kidney transplantation in July 2002, split liver transplantation (SLT) in April 2004, the first domino liver transplantation (DLT) in August 2005. Pediatric living donor liver transplantation (LDLT) was initiated in October 2006, adult LDLT was carried out in August 2007. In September 2007, the first living donor combined liver and kidney transplantation from the same donor in Asia was performed. The first domino+living donor double grafts liver transplantation in the world was performed in January 2009. In March 2011, we performed laparoscopically assisted right hepatic lobe liver transplantation (LDLT) with middle hepatic vein. In May 2014, living donor laparoscopic left lateral lobe procurement was successfully established. In April 2016, simultaneous liver, pancreas and kidney multi-organ transplantation was completed. Domino donor-auxiliary liver transplantation was performed in February 2017. In December 2017, extracorporeal membrane oxygenation (ECMO)-supported liver transplantation in a patient with severe pulmonary hypertension was successfully completed. Liver transplantation combined with partial splenectomy was established in April 2018. Cross-domino liver transplantation (hypersensitive kidney transplantation with auxiliary liver transplantation+pediatric liver transplantation) was performed in May 2018. During the 20 years, the team has performed or assisted other centers in Beijing, Shanghai, Guangzhou and Shenzhen to carry out more than 10 000 cases of liver transplantations. A total of 7 043 cases of various types of liver transplantation were performed in the single center of the hospital (6 005 adult liver transplantations and 1 038 pediatric liver transplantations). Concerning adult liver transplantation, the cumulative 1-year, 3-year and 5-year survival rate from September 1998 to March 2003 were 83.1%, 73.0% and 69.0%, from April 2003 to March 2009 were 85.3%, 76.2% and 72.1% and from April 2009 to September 2018 were 87.5%, 79.2% and 75.1%, respectively. The cumulative 1-year, 3-year and 5-year survival rate for pediatric liver transplantation were 93.5%, 92.2% and 90.2%, respectively. The nucleoside (acid) analogue combined with low dose hepatitis B immunoglobulin (HBIG) was developed to prevent the recurrence of hepatitis B after liver transplantation, this plan has reduced the recurrence rate of hepatitis B and the 5-year re-infection rate of hepatitis B virus (HBV) after liver transplantation significantly. The risk assessment system for tumor recurrence after liver transplantation was established and individual treatment method was established based on this assessment system. Continuous exploration and improvement of liver transplantation for liver cancer, liver re-transplantation, liver transplantation with portal vein thrombosis, SLT, DLT and multi-organ combined transplantation have significantly improved the clinical efficacy of patients and the post-operative survival rate. CONCLUSIONS The liver transplantation team of Tianjin First Center Hospital has carried out a scientific and technological exploration on the key problems and technical difficulties of clinical liver transplantation. This work strongly has initiated and promoted the rapid development of liver transplantation in China. The restrictive barrier of hepatitis B recurrence after liver transplantation has been overcome. The risk prevention and control system of tumor recurrence after liver transplantation has been established. A series of innovative achievements that can be popularized have been achieved in the field of complex liver transplantation and expansion of donor liver source. The iterative progress and sustainable development of liver transplantation have been realized.
China ; Humans ; Liver Transplantation

Esophageal and gastric varices are common complications of liver cirrhosis and are seen in 50% of patients with liver cirrhosis. The annual incidence rate of esophagogastric variceal bleeding is 5%-15%, and even if the recommended treatment is used, the 6-week mortality rate is still as high as 15%-20%. Spontaneous bacterial peritonitis (SBP) is a common complication of end-stage liver disease and has an incidence rate of 10%-30% in patients with severe liver damage. SBP refers to the bacterial infection of the peritoneum and/or ascites that occurs in the absence of any inflammation in adjacent tissues (e.g., intestinal perforation and intestinal abscess). Hepatic encephalopathy (HE) is the clinical syndrome manifesting as cognitive impairment in patients with chronic liver disease, and its pathogenesis has not yet been fully elucidated and may be associated with ammonia poisoning theory, γ-aminobutyric acid and endogenous benzodiazepine complex receptor theory, and inflammatory pathway theory. This article introduces the advances in the treatment of upper gastrointestinal bleeding in patients with liver cirrhosis, SBP, and HE in 2016.

Objective To summary and explore the indications,safety and effectiveness of no touch ostial cardioplegia delivery method during aortic valve replacement(AVR) for severe aortic insufficiency(AI),in order to provide guidance for clinical application.Methods Between September 2012 and February 2015,63 cases were randomly divided into two groups,in study group we used this technique in 31 patients with a mean age of(58.0 ± 15.0) years and a range of 31 to 74 years old.The degree of AI averaged (3.5 ± 0.5) according to a semiquantitative 5-grade scale[1].32 patients were enrolled in control group with a mean age of(60.4 ± 9.4) years and a range of 22 to 73 years old.The degree of AI averaged (3.6 ± 0.5).The abnormal aortic cusps were sutured though the aortic incision,which would be closed temporarily.Blood cardioplegia was infused repeatedly into the coronary osita through the closed aortic incision during the remainder of the procedure except AVR,when the closed aortic incision was reopened.Results In the study group,spontaneous resumption of nodal and normal sinus rhythm shortly after the removal of aortic cross-clamp was achieved in all patients,no hearts had to be electrically defibrillated and electrocardiogram showed no obvious S-T segment elevation.The time from cross-clamp removal to sinus rhythm was(12.3 ± 7.7) minutes in the study group,versus (18.6 ± 8.2) minutes in the control group(P =0.037).No operative mortality occurred in both groups.There was a significant increase in left ventricular ejection fraction[from(0.54 ± 0.09) preoperatively to(0.62 ± 0.06) postoperatively] (P =0.005) in patients of the study group,which was not seen in patients of the control group.Conclusion Preliminary results have shown that the no touch ostial cardioplegia delivery method is a feasible method for cardioplegia infusion during aortic valve replacement in patients with severe AI.

To optimize the water extraction process for glycyrrhiza. Methods: HPLC was used to determine the con-tents of glycyrrhizic acid and liquiritin. The comprehensive index included the contents of glycyrrhizic acid and liquiritin and the yield of dry extract. The water amount and the extraction time were selected as the independent variables, and the comprehensive index was set as the dependent variable. Design-expert 8. 06 software was used to fit multivariate linear or quadratic multinomial models for the experimental values. Response surface methodology was used to optimize the water extraction process. The prediction was carried out through comparing the observed and predicted values. Results:The regression coefficient of binomial fitting complex model was as high as 0. 979 7. The optimum conditions of extraction process were as follows:12-fold amount of water, extracting 3 times with 90 min for each time. The deviation between the observed and predicted values was -1. 72%. Conclusion: Central composite design-response surface methodology is convenient and highly predictive in optimizing the water extraction process for glycyrrhiza, which can be applied in the further membrane separation and purification.

OBJECTIVETo investigate the histogenesis, clinicopathologic characteristics, pathologic diagnosis, differential diagnosis, treatment and prognosis of basal cell carcinoma of the Prostate (BCCP).

METHODSWe conducted clinicopathologic analysis on the manifestations of 5 cases of BCCP by HE staining and immunohistochemistry and reviewed relevant literature.

RESULTSMicroscopically, the tumor cells were small in volume with ovoid karyomegaly and mitosis, some arranged like a solid nest or with a cribriform appearance. The tumors displayed an invasive growth, with positive expressions of 34betaE12, P63 and Cytokeratin 14, and negative expressions of PSA and P504s. No recurrence and metastasis were found in these patients during over 12 months of follow-up after surgery.

CONCLUSIONBCCP is a rare neoplasm different from prostate adenocarcinoma. Immunohistochemistry is indispensable in distinguishing this tumor from other types of prostatic carcinoma. Its biological behavior remains to be further studied. The best treatment option may be radical surgery combined with chemo-radiotherapy at the present time.

Aged ; Carcinoma, Basal Cell ; pathology ; Humans ; Male ; Middle Aged ; Prostatic Neoplasms ; pathology

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Meningioma ; diagnosis ; Middle Aged ; Retrospective Studies ; Young Adult

BACKGROUNDMeningioma is one of the most common primary tumors of the central nervous system, but there are not many detailed studies on the sex, age, subtypes and locations of large series. This study was a retrospective analysis of the characteristics of meningioma cases consecutively operated on at a single institution in China from 2001 to 2010.

METHODSThis study investigated the demographic background of 7084 meningioma cases, and the subtypes and locations of the tumors. Sex and age distributions were analyzed, and the pathological subtypes were classified according to the World Health Organization (WHO) classification. The location of the meningiomas was also categorized.

RESULTSThe female:male ratio of the 7084 cases was 2.34:1. The mean age was 51.4 years (range, 11 months-86 years). The mean age of cases of WHO grade I meningioma was significantly older than that of grade II or III meningiomas (P < 0.001, Fisher's Least Significant Digit test). There was a significantly higher female:male ratio in WHO grade I meningiomas than in grade II or grade III meningiomas (2.57, 1.03 and 0.76, respectively; P < 0.001, χ(2) test). Meningothelial (n = 2061) and fibrous meningiomas (n = 3556) were the most common subtypes, comprising 79.3% of all meningiomas. All meningioma cases were classified into 23 locations in this study, with the cerebral convexity the most common site (38.33%, n = 2722). Cases with uncommon locations such as extra-cranial and sylvian fissure meningiomas were also present in this series.

CONCLUSIONSFemale predominance was found for benign meningiomas, while malignant subtypes showed male predominance. The mean age of patients with WHO grade I meningiomas was older than that of patients with higher-grade tumors. Meningothelial and fibrous meningiomas were the most common subtypes. The cerebral convexity was the most common meningioma location.

Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Meningioma ; epidemiology ; Middle Aged ; Sex Distribution ; Young Adult

OBJECTIVETo identify a novel human leukocyte antigen (HLA) B allele and explore its family heritage.

METHODSA novel HLA allele was suspected upon routine HLA typing using a polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) assay. The sequence was confirmed with DNA sequencing and compared with its closest matching allele, B*55:02. The family was also investigated.

RESULTSAn unusual reaction pattern was detected during routine HLA typing. The sequence was confirmed to be a novel HLA-B allele, which differed from the closest matching allele, B*55:02 in 7 nt positions in exon 2. Among the 7 mutations from 6 codons, there were two amino acids changes including 69Glu→Met and 70Glu→Ala.

CONCLUSIONA novel HLA-B allele has been identified and officially named as B*55:35 by the WHO Nomenclature Committee for Factors of the HLA System (GenBank accession number FJ898284).

Alleles ; Base Sequence ; HLA-B Antigens ; genetics ; Histocompatibility Testing ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA