China ; Critical Care ; history ; methods ; trends ; History, 20th Century ; History, 21st Century ; Humans

HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication

Objective To summarize the clinical characteristics and diagnosis and treatment of hemangioma of small intestine in children.Methods Location,pathology,clinical manifestation,diagnosis and treatment of 3 cases from our hospital and 44 cases reported in our courtry with hemangioma of the small bowel were analyzed from 1994 to 2004.Results These tumors locating in jejunum,ileum,duodenum as well as parts small intestine were 31.9%,42.6%,4.2% and 21.3%,respectively.Solitary and multiple tumors were 29.8% and 70.2% separately.The histopatholical report of 10 cases revealed that capillary,cavernous,mixed type hemangioma and hematolymphangioma were 2,6,1 and 1 cases,respectively.Thirty nine children presented with recurrent black stool.Five patients manifest in the form of intussusception.One child passed a bloody stool so massive as to cause shock.Small-bowel obstruction occurred in 5 cases.The hemoglobin of 38 patients were lower than 90 g/L.The incidence of preoperative diagnosis was only 10.6%.All of patients were performed operation.Conclusions The frequent locations of hemangioma of small intestine are in ileum and jejunum,and multiple tumors are common.The diseases are characterized as recurrent hematochezia with painlessness.Preoperative diagnosis of a small bowel hemangioma can be very difficult.The segments of small bowel with hemangioma resection are the most method of the treatment.

Objective: To analyze the efficiency of ureteroscope and biopsy in the diagnosis of tumor grade, muscle-invasiveness and multifocality in suspected upper tract urinary carcinoma (UTUC) patients in order to find out whether it can be used in the risk stratification of UTUC patients.Methods: A retrospective study of 76 UTUC patients who underwent preoperative ureteroscope and/or biopsy and received radical nephroureterectomy in Peking University Third Hospital during January 2014 to December 2016 was undertaken.Results: In this study, 76 patients were included.There were 31 males (40.8%), and 45 females (59.2%).The median age was 64.5 years (31-88), and 51 patients had the symptom of hematuresis.The tumor was located in renal pelvis in 39 patients, and in ureter in 37 patients.Post-operative pathology confirmed that all the 76 patients included in this study suffered from UTUC, of whom 21 (21.6%) were of low-grade, 51 (67.1%) were of high-grade, 4 (5.3%) were undetermined, and 47 (61.9%) patients were muscle-invasive, and 27 (35.5%) were not, and 2 (2.6%) were undetermined.Among the 50 patients, in whom the grade of the tumor could be diagnosed by biopsy, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value for low-grade tumor was 88.2%, 69.7%, 76.0%, 60.0% and 92.0%, respectively.Among the 27 patients, in whom the muscle-invasiveness could be diagnosed by biopsy, 5 patients were diagnosed with muscle-invasiveness, all confirmed by pathology after surgery and 22 patients were diagnosed with none muscle-invasiveness, turned out to be 50% muscle-invasive and 50% none-muscle invasive after surgery.The accuracy was 59.3%.The accuracy of ureteroscopic biopsy to diagnosis multifocality was 61.0%.On univariate ana-lysis, biopsy grade was associated with postoperative pathology (P=0.001), while gender, age, side, body mass index (BMI), hematuresis, preoperative estimated glomerular filtration rate (eGFR), hydronephrosis, tumor size, location, multifocality and sessile were not associated with postoperative pathology grade.Biopsy grade (P=0.02), preoperative eGFR<90 mL/(min·1.73 m2)(P=0.025) and tumor located in pelvis (P=0.049) were associated with muscle invasiveness.Gender, age, side, BMI, hematuresis, hydronephrosis, tumor size, multifocality and sessile were not significantly associated with muscle invasiveness.Conclusion: Ureteroscope and biopsy can assist risk stratification in upper tract urothelial carcinoma patients.

Objective To characterize the audiological features in the infants with otitis media with effusion(OME)and to investigate the utility of variety of objective andiometry methods in diagnosis and intervention on OME.Methods Fifry six infants(40 males and 16 females)were investigated,who were referred to our clinic at the General Hospital of Chinese People's Liberation Army by the other hospitals from December 2004 to June 2007 when the infants were diagnosed or highly suspected of OME.The ages at the initial diagnosis ranged from 42 days to three years,with an average of five months.The infants,after receiving the conventional otolaryngological exams,were subjected to the tests of auditory brainstem response (ABR),otoacoustic emission(OAE),tympanometry(226 Hz and 1000 Hz)and behaviors audiometry.Results Among 56 affected infants.87 ears were diagnosed with OME,of which 31 infants were affected bilateral and 25 with monaural.For the 49 infants who received hearing screening at birth.36 infants were referred at the initial screening.For the 52 infants who received repeated screening.all subjects were referred.Six infants without receiving hearing screening came to clinic when their parents observed their kids'hearing impairment.Among the 52 cases(104 ears)who received tympanometry test,20 subjects (28 ears)showed B or C type tympanometry curve.Thirty-nine cases(78 ears)were given tympanometry test at 1000 Hz,of which 38 cases(55 ears)showed abnormal hearing.Among 56 infants(112 ears)with ABR test,49 subjects(74 ears)exhibited prolonged ABR type Ⅰ curve.All 56 infants(112 ears)received OAE test,of which 55 subjects(81 ears)were referred Four infants(8 ears)accepted the behavior test and all of them showed A-B Gap.Conclusions The combined tympanometry test at both 226 Hz and 1000 Hz,ABR latency or threshold test,infant's behavior test and OAE,used jointly,enable characterizing better OME in infants.thus helping early diagnosis of this hearing disorder.

To investigate JAK2V617F mutation and its clinical significance in patients with idiopathic myelofibrosis (IMF), genomic DNA was extracted from peripheral blood cell samples of 12 IMF cases. Allele-specific PCR (AS-PCR) was performed to identify JAK2V617F mutation, and the results were confirmed by sequence analysis. A retrospective study was performed to explore the correlation between JAK2V617F mutation and the clinical, hematologic features. The results showed that in follow-up for 2 to 15 months, the occurrence of the positive point mutation in 12 patients with IMF was 50%, and the half of these positive patients had thrombosis. Patients with JAK2V617F point mutation had a higher counts of platelets and megakaryocytes in bone marrow than those in patients without JAK2V617F point mutation. Out of other 6 IMF patients without JAK2V617F point mutation only 1 patient had thrombosis, and lower counts of platelets in peripheral blood and megakaryocytes in bone marrow. It is concluded that majority of IMF patients with positive JAK2V617F point mutation have typical clinical and hematologic features, higher incidence of thrombosis, and higher counts of platelets in peripheral blood and megakaryocytes in bone marrow.
Adult ; Aged ; Base Sequence ; Bone Marrow ; pathology ; Female ; Follow-Up Studies ; Humans ; Janus Kinase 2 ; genetics ; Male ; Megakaryocytes ; pathology ; Middle Aged ; Molecular Sequence Data ; Platelet Count ; Point Mutation ; Primary Myelofibrosis ; genetics ; Retrospective Studies

Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Remission Induction ; fms-Like Tyrosine Kinase 3

OBJECTIVETo investigate the effect of hyperthermic intraoperative intraperitoneal chemotherapy(HIIC) and postoperative nutritional support on the intestinal permeability and the cellular immunity function in patients with advanced gastric cancer.

METHODSAll the patients diagnosed as advanced gastric cancer in the Gastric Tumor Diagnosis and Treatment Center of Jiangsu Cancer Hospital were randomly divided into three groups using random digit table:(1)EN group treated with enteral nutrition during postoperative period; (2)HIIC+EN group treated with HIIC combined with postoperative enteral nutrition;(3)HIIC+PN group treated with HIIC combined with postoperative parenteral nutrition. Index of lactulose/mannitol(L/M) ratio was used to evaluate the permeability of intestinal mucosa. The percentage of CD4(+), CD8(+) and NK cell, the ratio of CD4/CD8 T cell in peripheral blood were tested by flow cytometry. The time points of these measurements were the day before surgery, postoperative days (POD) 3, 7, and 12.

RESULTSCompared with the day before surgery(POD-1), the ratio of L/M on POD+3 increased significantly in all the three groups(0.1235±0.0611 vs. 0.0280±0.0183, 0.1648±0.0571 vs. 0.0305±0.0208, 0.1702±0.0628 vs. 0.0298±0.0229)(P<0.05) and then decreased gradually. The L/M ratio of EN(0.0278±0.0217) and HIIC+EN(0.0336±0.0235) groups recovered to the baseline on POD+12, however HIIC+PN group still had elevated L/M ratio(0.0616±0.0430). The percentage of CD4(+)T cell and the ratio of CD4/CD8 in HIIC+EN group and HIIC+PN group were significantly lower than those in EN group(P=0.033, P=0.002, respectively). Compared with the POD-1,the percentage of CD4(+)T cell and the ratio of CD4/CD8 in HIIC+EN group and EN group on POD+12 were increased significantly(P<0.05).

CONCLUSIONSHIIC may cause significant increase in intestinal permeability and inhibit cellular immunity in patients undergoing radical resection for advanced gastric cancer. Mucosal permeability can be reversed by enteral nutrition.

Abdominal Cavity ; Adult ; Aged ; Chemotherapy, Cancer, Regional Perfusion ; Female ; Humans ; Immunity, Cellular ; Intestinal Mucosa ; immunology ; physiopathology ; Intraoperative Care ; Male ; Middle Aged ; Nutritional Support ; methods ; Permeability ; Postoperative Period ; Stomach Neoplasms ; immunology ; physiopathology ; therapy

OBJECTIVESTo study the effect of testicular local heating on spermatogenic cell apoptosis in rat.

METHODSSeventy male SD rats were divided into heat treatment group (43 degrees C) and control group (22 degrees C). Each group was further divided into seven sub-groups respectively according to the time of 12 hours and 1 days, 3 days, 6 days, 10 days, 50 days and 80 days after testicular local treatment. The spermatogenic cell apoptosis in all sub-groups was examined by means of electron microscopy, flow cytometry and terminal deoxynucleotidyl transferase-mediated dUDP-nick end labeling(TUNEL) method.

RESULTSIn the groups of heat treatment, spermatogenic cell apoptosis was detected by electron microscopy; flow cytometry showed that the percentage of cells with sub-haploid increased(P < 0.01); the percentage of positive TUNEL cells in the heat treatment groups was higher than that in the control group(P < 0.01). Initiation of spermatogenic cell apoptosis after testicular heating was not random but was highly selective.

CONCLUSIONSLocal testicular heating could increase the spermatogenic cell apoptosis. The most sensitive cell is spermatocyte. Spermatid and sperm also display apparent changes. Heating can increase the apoptosis of spermatogonia in a long period.

Animals ; Apoptosis ; Flow Cytometry ; Hot Temperature ; In Situ Nick-End Labeling ; Male ; Microscopy, Electron ; Rats ; Rats, Sprague-Dawley ; Spermatogonia ; cytology ; ultrastructure ; Testis ; pathology ; ultrastructure

OBJECTIVE: To profile urinary metabolite variations from 1, 2-dimethylhydrazine (DMH)-induced precancerous colon rats, Jinfu Kang treated rats and healthy controls.

METHODWe used ethyl chloroformate derivatization and gas chromatography-mass spectrometry (GC-MS) based metabonomic method to analyze rat urines.

RESULTThe time-dependent variations of metabolite profile showed a progressive deviation of the metabolism in the model group from the initial pattern over time and a systemic recovery of the metabolism in the treatment group, which is consistent with the histological results. The in-depth analysis indicated that the disorder of tricarboxylic acid cycle (TCA), tryptophan metabolism, polyamine metabolism and gut flora structure were associated with DMH intervention.

CONCLUSIONMetabolic study revealed that Jinfu Kang can effectively reverse metabolic departures in DMH-induced precancerous colon rat, which is consistent with pathological results.

Animals ; Colonic Neoplasms ; chemically induced ; pathology ; Colonic Polyps ; chemically induced ; drug therapy ; urine ; Dimethylhydrazines ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Wistar