Myostatin gene (MSTN) encodes a negative regulator for controlling skeletal muscle growth in animals. In this study, MSTN^-/- homozygous mutants with "double muscle" phenotypic traits and stable inheritance were bred on the basis of MSTN gene editing rabbits, with the aim to establish a method for breeding homozygous progeny from primary MSTN biallelic mutant rabbits. MSTN^-/- primary mutant rabbits were generated by CRISPR/Cas9 gene editing technology. The primary mutant rabbits were mated with wild type rabbits to produce F1 rabbits, whereas the F2 generation homozygous rabbits were bred by half-sibling mating or backcrossing with F1 generation rabbits of the same mutant strain. Sequence analysis of PCR products and its T vector cloning were used to screen homozygous rabbits. The MSTN mutant rabbits with 14-19 week-old were weighed and the difference of gluteus maximus tissue sections and muscle fiber cross-sectional area were calculated and analyzed. Five primary rabbits with MSTN gene mutation were obtained, among which three were used for homozygous breeding. A total of 15 homozygous rabbits (5 types of mutants) were obtained (M2-a: 3; M2-b: 2; M3-a: 2; M7-a: 6; M7-b: 2). The body weight of MSTN^-/- homozygous mutant rabbits aged 14-19 weeks were significantly higher than that of MSTN^+/+ wild-type rabbits of the same age ((2 718±120) g vs. (1 969±53) g, P < 0.01, a 38.0% increase). The mean cross sections of gluteus maximus muscle fiber in homozygous mutant rabbits were not only significantly higher than that of wild type rabbits ((3 512.2±439.2) μm² vs. (1 274.8±327.3) μm², P < 0.01), but also significantly higher than that of MSTN^+/- hemizygous rabbits ((3 512.2±439.2) μm² vs. (2 610.4±604.4) μm², P < 0.05). In summary, five homozygous mutants rabbits of MSTN^-/- gene were successfully bred, which showed a clear lean phenotype. The results showed that the primary breeds were non-chimeric mutant rabbits, and the mutant traits could be inherited from the offspring. MSTN^-/- homozygous mutant rabbits of F2 generation could be obtained from F1 hemizygous rabbits by inbreeding or backcrossing. The progenies of the primary biallelic mutant rabbits were separated into two single-allelic mutants, both of which showed a "double-muscle" phenotype. Thus, this study has made progress in breeding high-quality livestock breeds with gene editing technology.
Animals ; CRISPR-Cas Systems/genetics* ; Gene Editing ; Muscle, Skeletal/metabolism* ; Mutation ; Myostatin/metabolism* ; Phenotype ; Rabbits

Objective:To investigate the efficacy and safety of ticagrelor and clopidogrel in patients with coronary atherosclerotic heart disease (CHD) and hyperhomocysteinemia (Hhcy).Methods:A total of 101 patients with CHD complicated with HHcy who had successfully undergone percutaneous coronary intervention (PCI) were enrolled. They were divided into ticagrelor group ( n=49) and clopidogrel group ( n=52) according to the different antiplatelet drugs used. The clinical data of the two groups were collected, and the incidence of major cardiovascular and cerebrovascular adverse events (MACCE) and bleeding events in one year were compared between the two groups. Results:There was no statistical difference in baseline datas and PCI datas between the clopidogrel and ticagrelor groups ( P>0.05). Compared with clopidogrel group, ticagrelor can reduce the total MACCE (8.16% vs 32.69%, P<0.05) and the incidence of unstable angina pectoris events (0 vs 13.46%, P<0.05) in patients with hyperhomocysteinemia PCI 1 year after operation. The incidences of ischemic stroke, unstable angina pectoris, recurrent myocardial infarction, and cardiogenic death were compared separately between the two groups, and the difference was not statistically significant ( P>0.05). The incidence of major bleeding events, minor bleeding events, and minimal bleeding events were similar between the two groups, and the difference was not statistically significant (4.08% vs 0; 4.08% vs 1.92%; 20.41% vs 9.62%; P>0.05). Conclusions:In patients with CHD and Hhcy, ticagrelor can play a better antithrombotic effect, reduce the incidence of ischemic events, and has good security, which is more worthy of clinical recommendation.

Objective:To compare the clinical and prognostic characteristics of ovarian endometrioid carcinoma (OEC) patients with synchronous endometrial lesions and patients with pure OEC.Methods:A retrospective review of the medical records of patients received initial treatment and a postoperative pathological diagnosis of OEC at Peking University People′s Hospital between August 1998 and December 2017 were performed. According to the inclusion criteria, a total of 56 patients with OEC were included in the study, including 13 patients concurrent with simultaneous endometrial lesions (Group A) and 43 patients with pure OEC (Group B).Results:Patients with synchronous endometrial lesions accounted for 23% (13/56). Mean age of Group A at diagnosis was (44.9±8.3) years old, 2/13 of patients were postmenopausal, and no one had a history of hypertension, the first symptom of 5/13 people was irregular vaginal bleeding. Mean age of Group B patients at diagnosis was (52.7±10.2) years old, 53% (23/43) of patients were postmenopausal, and 28% (12/43) patients had the history of hypertension, the first symptom of 4 (9%, 4/43) people was irregular vaginal bleeding. The differences of age, menopause status, history of hypertension and initial symptoms between the two groups were statistically significant (all P<0.05). There were no significant differences in fertility history, dysmenorrhea history, age of menarche, history of endometriosis, preoperative and postoperative CA 125 level, International Federation of Gynecology and Obstetrics (FIGO) stage, tumor grade, metastatic site and platinum-based chemotherapy drug resistance between the two groups (all P>0.05). The overall 5-year survival rate of OEC patients was 91.6%, and the overall 5-year progression-free survival rate was 76.6%. Among them, the 5-year survival rate of the OEC concurrent with simultaneous endometrial lesions group was 80.2%, and the pure OEC group was 93.4%; the 5-year progression-free survival rate of the OEC concurrent with simultaneous endometrial lesions group was 74.1%, and the 5-year progression-free survival rate of the pure OEC group was 77.3%. There were no significant differences between the two groups (all P>0.05). Multivariate analysis showed that the independent factors for the prognosis of OEC patients were FIGO stage ( P=0.006) and residual lesion size ( P=0.020). Conclusions:OEC patients have a high proportion of simultaneous endometrial lesions. OEC with simultaneous endometrial lesions are younger than patients with pure OEC. Synchronous endometrial lesions do not affect the prognosis of patients with OEC.

The article introduces the content and organization of the training program for newly recruited nurses in the three hospitals of Massachusetts General Hospital, Mayo Clinic, and University of Illinois Chicago Medical Center in the United States. This article also summarizes the characteristics and highlights of the program implementation, and discusses and analyzes the training program by combining the current situation of newly recruited nurses training in my country, and makes suggestions for further improvement in the future.

Objective:To evaluate the efficacy and safety of endoscopic trans-gastric gallbladder-preserving polypectomy in treatment of gallbladder polyps.Methods:A retrospective analysis was performed on data of 25 patients with gallbladder polyps who underwent endoscopic trans-gastric gallbladder-preserving polypectomy in the endoscopy center of Zhongshan Hospital from September 2017 to August 2019. Completion of operation, operation time, postoperative hospital stay, complications and follow-up results of patients were obtained and analyzed.Results:There were 13 cases of single polyp and 12 cases of multiple polyps, 13 cases of simple gallbladder polyps, and 12 cases of polyps and gallstones. The maximum diameter of polyps was 0.2-1.5 cm. The procedure failed in 1 patient (4%) because of the severe adhesion of abdominal cavity and disappearance of gallbladder when passing through the gastric wall. Twenty-four patients (96%) were successfully treated with endoscopic trans-gastric gallbladder-preserving polypectomy. The operation time was 67.1±26.8 min (35-140 min). The median hospital stay was 4 d (2-5 d). No severe adverse events including delayed bleeding, delayed perforation, diffuse peritonitis or abdominal abscess occurred. During the median follow-up time of 8 months (0-23 months), no patient was lost, no recurrence of polyps and no severe adverse events related to metal clips occurred.Conclusion:Endoscopic trans-gastric gallbladder-preserving polypectomy shows good short-term efficacy and is technically feasible, however, long-term outcome still requires further research.

Objective:To summarize the surgical treatment experience of thoracic solitary fibrous tumors(SFTs),especially some spe-cial SFTs.Methods:Patients with SFTs in our department between January 2009 and May 2017 were recruited in this study and the clinical characteristics were summarized.Results:Twenty-one patients(11 men and 10 women)with an average age of 48.3 years were included.Tumor sizes ranged from 3 to 30 cm and 13 patients(61.9%)harbored pleural tumors,while 8 patients harbored tu-mors from other special thoracic sites (including 1 case of trachea SFT and 1 case of pulmonary SFT). Surgical treatment was per-formed in 19 patients,17 of whom underwent complete tumor resection and two patients underwent palliative resection.One patient did not undergo surgery and was lost to follow-up,and another patient who did not undergo surgery died because of a heart attack af-ter discharge.The 19 patients who underwent surgery were followed up for 1-96 months,3 patients of whom underwent reoperation for tumor recurrence at 12,26,and 72 months after surgery,respectively,while the remaining patients had no recurrence.Seven pa-tients(33.3%)were confirmed with malignancy after operation or biopsy.Conclusions:Thoracic SFTs are mainly from the pleura and the surgical results are favorable.Resection after recurrence can achieve favorable results.

OBJECTIVETo explore the genetic basis for a Chinese pedigree affected with congenital hypofibrinogenamia.

METHODSPeripheral blood samples were collected from 9 members from the pedigree. Routine coagulation tests including activated partial thromboplastin time (APTT), thrombin time (TT), the prothrombin time (PT) were carried out. The activity of fibrinogen (Fg: C) was measured using Clauss method, and fibrinogen antigen (Fg: Ag) was measured with immunoturbidimetry. All exons and exon-intron boundaries of the fibrinogen Aα, Bβ and γ chain genes were amplified using PCR, which was followed by direct sequencing. Suspected mutation was confirmed by reverse sequencing. The mutant fibrinogen was analyzed with Swiss-PdbViewer.

RESULTSThe proband showed prolonged APTT, PT and TT. Her functional fibrinogen (Fg: C) and antigen fibrinogen (Fg: Ag) levels were reduced to 0.69 g/L and 0.72 g/L, respectively. Her mother and grandmother also had a low levels of fibrinogen, which were 0.99 g/L and 0.83 g/L for Fg: C, 1.02 g/L and 0.87 g/L for Fg: Ag, respectively. The results of other members from the pedigree were all within the normal range. Genetic analysis reveled a heterozygous G>T mutation at nucleotide 7590 in exon 8 of γ gene in the proband, which was predicted to be a novel Ser313Ile mutation. The mutation was also found in her mother and grandmother. Model analysis showed that the Ser313Ile mutation disturbed the hydrogen bonds between Ser313, Asn319 and Asp320. Moreover, the mutation also altered the mutual electrostatic force and affected the folding and instability of the mutant fibrinogen.

CONCLUSIONThe heterozygous Ser313Ile mutation probably underlies the hypofibrinogenemia in this pedigree.

Adult ; Afibrinogenemia ; genetics ; Female ; Fibrinogen ; chemistry ; genetics ; Heterozygote ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree

We present a case of two-year old boy with solitary atypical adenomatous hyperplasia (AAH) in extralobar sequestration (ELS), which was misdiagnosed as diaphragmatic hernia before surgery. Review of AAH and pulmonary sequestration (PS) revealed that the present case is the youngest of solitary AAH and also the first report of solitary AAH arising in ELS without a primary lung cancer. In a sense, the present case firstly supports the hypothesis that ELS may be an underlying cancer predisposition syndrome, so aggressive surgical therapy should be recommended for ELS.

OBJECTIVETo explore the phenotype, genotype and molecular mechanism for two pedigrees affected with hereditary antithrombin (AT) deficiency.

METHODSClinical diagnosis was validated by assaying of coagulation parameters including prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, antithrombin activity (AT:A) and specific antigen (AT:Ag), protein C activity, as well as protein S activity. To detect potential mutations in the probands, all exons, exon-intron boundaries and the 3', 5' untranslated regions were amplified by PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and silver staining. The effect of mutations on the AT protein was analyzed with bioinformatics software.

RESULTSThe AT:Ag of pedigree 1 was normal, but its AT:A has reduced to 30%. A heterozygous c.235C>T mutation in exon 2 causing p.Arg47Cys, in addition with two single nucleotide polymorphisms (c.981G>A, c.1011G>A) in exon 5 were identified in the patient. His four children, except for the elder daughter, were heterozygous for the mutations. The plasma levels of AT:A and AT:Ag in proband 2 have decreased to 39% and 103 mg/L, respectively. A heterozygous deletion (g.5890-5892delCTT) leading to loss of p.Phe121 was also detected in his father. Bioinformatic analysis suggested that the missense mutation Arg47Cys can affect the functions of AT protein. Meanwhile, lacking of Phe121 will result in loss of hydrogen bonds with Ala124, Lys125 and the cation π interactions with Lys125, Arg47, which may jepordize the stability of the protein.

CONCLUSIONThe proband 1 had type II AT deficiency, while proband 2 had type I AT deficiency. The p.Arg47Cys and g.5890-5892delCTT mutations of the AT gene are significantly correlated with the levels of AT in the two probands, respectively.

Adult ; Aged, 80 and over ; Antithrombin III ; genetics ; metabolism ; Antithrombin III Deficiency ; enzymology ; genetics ; physiopathology ; Exons ; Female ; Genetic Testing ; Genotype ; Humans ; Male ; Mutation ; Partial Thromboplastin Time ; Pedigree ; Phenotype ; Protein C ; genetics ; metabolism ; Protein S ; genetics ; metabolism

Objective To explore the effect of newly-designed gowns for critical patients. Methods Two hundred critical patients hospitalized for more than 3 days in the intensive care unit of our hospital during February 2013 to February 2015 were divided into the control group and observation group equally according to their odd or even registration number. The control group wore conventional dresses for patients (including a shirt and a pant) and the patients in the latter group were newly-designed gowns. The two groups were compared in terms of comfort, nursing manpower, nursing stuff and security. Result The observation group was significantly superior to the control group in terms of comfort, nursing manpower, nursing stuff and security (P<0.05). Conclusions The newly-designed gowns for critical patients can improve the comfort degree and save nursing manpower and stuff. It allows convenience in treatment and nursing.