Objective To investigate the feasibility of using clustered regularly interspaced short palindromic repeats ( CRISPR)-mediated genome editing to downregulate the expression of programmed cell death protein 1 (PD-1) on primary T cells by using a lentivirus delivery system. Methods Lentivirus vec-tors pLentiCRISPR A1-A6 containing different PD-1 genomic DNA sequences as single guide RNA ( sgRNA) for Cas9 targeting were constructed individually. The lentivirus vectors were tranduced into primary CD4 T cells. Flow cytometry analysis was performed to detect the expression of PD-1 for evaluating the knockout ef-ficiency. Results The lentivirus vectors pLentiCRISPR A1-A6 carrying six different target sites were con-structed and respectively tranduced into primary CD4 T cells. The expression of PD-1 accompanied with the activation of T cells. Co-expression of CD25 and PD-1 was observed on activated T cells. All of the six sites could be targeted by Cas9, of which A2 and A6 sites were more efficient in knocking out the gene encoding PD-1 with a rate of 19% and 29%, respectively. Conclusion This study suggests that it is feasible to knock out the expression of PD-1 on primary T cells by using CRISPR.

OBJECTIVETo explore the reasons of secondary fracture after percutaneous vertebroplasty (PVP) for osteoporotic vertebral compression fractures (OVCFs) and discuss the measure of prevention and cure.

METHODSFrom January 2011 to January 2013, the clinical data of 180 patients with primary OVCFs treated by PVP were retrospectively analyzed. There were 75 males and 105 females, aged from 68 to 95 years old with an average of (79.50 ± 5.45) years. The involved vertebrae were identified according to the clinical symptoms and imaging data. PVP were performed in 362 vertebrae and the patients were followed up with an average of 12 months. Subsequent vertebral fractures were found through the pain's reappearance and MRI or bone scan. The patients were divided into secondary fracture group and no-secondary fracture group according to the subsequent fractures or no. Secondary fracture group was divided into two groups according to gender, and the patients with secondary fracture were also categorized into the original surgical vertebral fractures, adjacent vertebral fracture and remote vertebral fractures. The age, gender, the cement volume, the cement leakage, secondary fracture site, the incidence and type of secondary fracture were observed and compared among different groups.

RESULTSAmong the 362 vertebrae of PVP, there were 109 vertebrae in male and 253 vertebrae in female. And 27 vertebrae (10 in male and 17 in female) of 22 cases (9 males and 13 females) occurred secondary fracture. The second PVP were performed in 13 cases (16 vertebrae) and the third PVP in 2 cases (4 vertebrae); 7 cases (7 vertebrae) were treated with conservative therapy. There was no statistically significant difference on age, gender, cement volume and leakage between secondary fracture group and no-secondary fracture group (P > 0.05). There was no statistically significant difference on the incidence and type of secondary fracture between male and female (P > 0.05). No significant difference was found on the adjacent and remote vertebral fractures (P > 0.05). Most of secondary fracture occurred in 6 months, and whether the single and double side injection, cement leakage had no obvious relation.

CONCLUSIONThere is no significant difference in the subsequent fracture after PVP for the OVCFs different gender and fractured site, and also no significant difference in the adjacent and remote vertebral fractures. The report didn't support the biomechanical viewpoint that vertebral body stiffness increasing after PVP would lead to adjacent vertebral stress increasing and result easily in adjacent vertebral fracture. Secondary fracture occurs always in 6 months after operation, which is the natural course of osteoporosis.

Aged ; Aged, 80 and over ; Female ; Fractures, Compression ; surgery ; Humans ; Male ; Osteoporotic Fractures ; surgery ; Postoperative Complications ; etiology ; Recurrence ; Retrospective Studies ; Spinal Fractures ; surgery ; Vertebroplasty

OBJECTIVETo explore the therapeutic effect and the mechanism of acupuncture in reducing body weight.

METHODSFifty cases of simple obesity were randomly divided into an acupuncture group and a drug control group, 25 cases in each group. The patients in the acupuncture group were treated with body acupuncture at Tianshu (ST 25), Guanyuan (CV 4), etc, electroacupuncture and ear point tapping and pressing therapy (Shenmen, Endocrine, etc. were selected); the patients in the control group were treated with oral administration of sibutramine. The serum leptin level before and after treatment were determined and the therapeutic effect in reducing body weight was assessed.

RESULTSThe total effective rate was 88.0% in the acupuncture group and 80. 0% in the control group, with no significant difference between the two groups (P>0.05); after treatment, the serum leptin level in both the two groups decreased significantly (P<0.01), and the decreasing amplitude in the acupuncture group was superior to that in the control group (P<0.05).

CONCLUSIONAcupuncture therapy has a definite therapeutic effect in reducing body weight, and the improvement of serum leptin level of the patient is possibly one of the mechanisms of acupuncture in reducing body weight.

Acupuncture Points ; Acupuncture Therapy ; Body Weight ; Electroacupuncture ; Humans ; Leptin ; blood ; Obesity ; therapy

Acquired Immunodeficiency Syndrome ; drug therapy ; genetics ; surgery ; Adult ; Burkitt Lymphoma ; drug therapy ; genetics ; surgery ; virology ; Diagnosis, Differential ; Female ; Genes, myc ; HIV ; isolation & purification ; HIV Infections ; Herpesvirus 4, Human ; genetics ; Humans ; Immunohistochemistry ; Lymphoma, AIDS-Related ; drug therapy ; genetics ; surgery ; virology ; Lymphoma, B-Cell ; pathology ; Lymphoma, Mantle-Cell ; pathology ; Male ; Middle Aged ; RNA, Viral ; analysis ; Sarcoma, Myeloid ; pathology ; Translocation, Genetic

OBJECTIVETo study the value of MRI in the diagnosis of cerebral sparganosis in children.

METHODSThe MRI appearances of 17 children with cerebral sparganosis proven by pathology or serological test were retrospectively studied. The diagnostic accuracy rate of cerebral sparganosis was compared before and after knowing the imaging features of this disease.

RESULTSEnhanced MRI was performed in 12 cases, and all of them demonstrated abnormal enhancement. Peripheral ring-type, tortuous beaded shape or serpiginous tubular shape enhancement were found in 8 cases. After analysis of follow-up MRI for 7 cases, a change in location and shape of lesions was found in 2 cases. Typical MRI patterns of cerebral sparganosis included abnormal enhancement such as peripheral ring-type, tortuous beaded shape or serpiginous tubular shape enhancement, and changes in location and shape of lesions in the follow-up MRI. The diagnostic accuracy rate for the 17 cases of cerebral sparganosis at the first visit, after first imaging examination and after several follow-up MR examinations were 0%, 11.8% and 28.6%, respectively. The diagnostic accuracy rate of this disease increased to 64.7%, 70.6% and 85.7% respectively after the radiologists were trained in the imaging characteristics of the disease.

CONCLUSIONSMR findings in children with cerebral sparganosis are specific. Mastering correctly the typical imaging features of the disease can be very helpful in reducing the rate of clinical misdiagnosis.

Adolescent ; Central Nervous System Helminthiasis ; diagnosis ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Retrospective Studies ; Sparganosis ; diagnosis ; pathology

Objective To explore the dosage and injection method of concanavalin A(Con A) for inducing Wistar rats into the acute hepatic injury model. Methods (1)According to the dosage of Con A, 42 Wistar rats were randomly divided into groups A, B, C, D, E, N, 7 rats in each group. Group N was given tail intravenous injection of normal saline as normal control group. Groups A, B, C, D, E were given intravenous injection of 4, 8, 16, 30, 40 mg/kg of Con A respectively. At the 8th hour after modeling, the levels of alanine transaminase(ALT), aspartate aminotransferase(AST), albumin(ALB), interleukin(IL)-2 , IL-10, interferon (IFN)-γ, and tumor necrosis factor(TNF)-αwere detected. And HE staining was used to observe the pathological feature of hepatic tissue. (2)According to the injection method of Con A, 21 Wistar rats were randomly divided into normal control group, intraperitoneal injection group and tail intravenous injection group, 7 rats in each group. The dosage of Con A for the rats in intraperitoneal injection group and tail intravenous injection group was 16 mg/kg. At the 8th hour after modeling, the levels of serum ALT, AST, and ALB were determined. Results The number of abnormal deaths in various dose Con A groups at the end of each experiment was 0 in groups A, B, C, and 2 in group D, and 7 in group E. A small amount of spotty necrosis, inflammatory cell infiltration, and hepatic lobule with almost integrity of structure were found in groups A, B, while obvious bridging-like necrosis was seen in groups C, D. Serum ALT, AST, and ALB levels in intraperitoneal injection group had no statistically significant difference as compared with the normal control group. Conclusion Tail intravenous injection of 16 mg/kg of Con A can be used to induce an acute immunological liver injury rat model successfully.

Diabetes is a clinical syndrome caused by a variety of factors. It often causes multiple systemdamage, leading to chronic progressive lesions of the eye, kidney, blood vessels, heart, nerves and otherorgans. At present, its etiology and pathogenesis are not fully clarified, and there is a lack of effectivecure. Further exploration of the molecular regulatory mechanisms that drive diabetes and itscomplications, identifying specific biomarkers and molecular therapeutic targets, is undoubtedly aneffective strategy to prevent the onset and development of diabetes and improve the quality of life ofpatients. Long non-coding RNA (lncRNA) is an important regulator of body normal activity and diseasedevelopment. Abnormal expression and mutation are one of the main causes of diabetes and many otherdiseases. Nuclear paraspeckle assembly transcript 1 (NEAT1) is a newly discovered lncRNA molecule inrecent years. It has attracted much attention because of its important regulatory role and diverse biologicaleffects in diabetes and its complications. This article summarizes the molecular regulation mechanism andrelated biological functions of lncRNA NEAT1 in diabetes and its complications in order to provide a newscientific reference for early prevention, diagnosis and molecular targeted therapy of diabetes.

Objective To observe the effects of rotating magnetic field on the tail-flick latency (TFL)and mechanical withdrawal threshold(MWT)of rats and analyze it from processing time and angular velocity.Methods Forty-eight female SD rats were randomly assigned to 8 groups(n=6),which would be determined by 8 different time points(before treatment,treating for 10,20,30 and 60 min,10,20 and 30 min after treatment).The magnetic plate rotated near the lumbar of the rat at an angular velocity of 7300 r/min and the TFL and MWT with yon Frey hair test were determined at each time point.The second experiment was carried out on 40 female rats at an angular velocity of 6300,6800,7300 and 7800 r/min,respectively;TFL and MWT were determined 30 min after treatment.The data were statistically analyzed.Results The first experiment was concluded as follows:compared with that before treatment,TFL was statistically increased at treating for 10,20,30 and 60 min(P<0.05).The TFL gradually became shorten after the treatment,but it was still significantly longer 10 and 20 min after the treatment as compared with that before treatment(P<0.05).MWT increased significantly at treating for 30 and 60 min,and 10 min after treatment as compared with that before treatment(P<0.05).The second experiment showed that TFL significantly went up as the angular velocity increased(P<0.05).Significantly difference on MWT was found between the angular velocity of 7800 and 6300 r/min.Conclusion Pain threshold can be increased by rotating magnetic field following the increase of processing time and the angular velocity at a certain range,indicating that the rotating magnetic field might be a potential non-invasive pain alleviating method.

OBJECTIVETo observe the clinical outcomes and differences among three surgical procedures for avulsion fracture of tibial intercondylar eminence.

METHODSFrom October 1995 to October 2005, 3 different procedures had been performed on 49 patients, which included open reduction and internal fixation (Group A, n = 17), arthroscopic reduction and internal fixation( Group B, n = 19) and limited open reduction and internal fixation assisted with arthroscopy (Group C, n = 13). All patients were followed up for 1 to 10 years (average 4. 6 years).

RESULTSFor the 3 groups, normal extension function were 35.5%, 16.0% and 38.0%, mild abnormal 35.5%, 11.0% and 23.0%, moderate abnormal 29.0%, 47.0% and 31.0%; and severe abnormal were 0,26.0% and 8.0%. Normal flexion were 82.0%, 78.0% and 84.0%, mild abnormal 12.0%, 11.0% and 8.0%, moderate abnormal 6.0%, 11.0% and 8.0%. The positive rate of Lachman's or anterior drawer test were 35.0%, 45.0% and 38.0%; McIntoshi test were 11.0%, 16.0% and 13.0% respectively for 3 groups. Lysholm' scale were average 98.6, 97.3 and 98.2; Tegner' scale were 6.6, 6.4 and 6.7. KT-2000 showed that anterior translation of tibial were 6.9, 7.1 and 6.6 mm; side to side difference were 11.4, 1.7 and 1.5 mm, except that statistically significant differences were found in extension function between group A and group B (P = 0.02). There were no any statistically significant differences in other aspects.

CONCLUSIONLimited open and properly over reduction and three dimensional as well as strong internal fixation assisted with arthroscopy should been attempted for the treatment of avulsion fracture of tibial intercondylar eminence.

Adolescent ; Adult ; Arthroscopy ; Child ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery ; Treatment Outcome

OBJECTIVETo search differentially expressed proteins in serum of patients with Crohn disease.

METHODSSerum protein samples from 4 patients with Crohn disease and 8 healthy adults were recruited cross-labeled with variant CyDye, and then followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTSThe 2-D electrophoresis results were compared between the Crohn disease patients and the healthy adults. The spot 1058 expression in serum of Crohn disease patients increased by 1.68 folds as compared with healthy adults (P<0.05). The protein was identified as haptoglobin by mass spectrometry.

CONCLUSIONUp-regulating expression of haptoglobin in serum of Crohn disease patients may play a role in disequilibrium of immunity system.

Adult ; Blood Proteins ; metabolism ; Case-Control Studies ; Crohn Disease ; blood ; Electrophoresis, Gel, Two-Dimensional ; Haptoglobins ; metabolism ; Humans ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization