Objective:To clone and express Mycobacterium tuberculosis(Mtb) Rv2460c gene(encoding ClpP2 protein),and evaluate the immunogenicity of its coding product. Methods: The recombinant plasmid of pET32a (+) vector-ClpP2 that H37Rv Rv2460c gene was cloned into the plasmid pET32a(+)vector,was transformed into E. coli BL21(DE3)and induced expression by IPTG,then purified by affinity chromatography. The recombinant protein was confirmed by SDS-PAGE and Western blot. The analysis of immunogenicity of Mtb ClpP2 and its epitope prediction were performed by bioinformatic methods. The antibody levels of polyclonal antibody titer against ClpP2 protein in rabbits and TB patients′ serum were detected by ELISA. Results: The recombinant ClpP2 protease was expressed as inclusion bodies in E. coli. The purity of purified protein was 93% by bandscan software analysis. The bioin-formatics analysis shows Mtb ClpP2 protein has multiple preponderant B cell and T cell epitopes. Rabbit antiserum titer was 1∶64 000;Serum anti-ClpP2 antibody levels in TB patients was higher than that in healthy control subjects. Conclusion:The recombinant ClpP2 protein was purified, and specific Rabbit anti-ClpP2 polyclonal antibody was prepared successfully. Experiment and bioinformatic information studies showed that Mtb ClpP2 protease has strong immunogenicity.

Objective To investigate the significance and distribution of lymphopenia and antilymphocyte antibody(ALA) in systemic lupus erythematosus (SLE) and to explore the role of ALA in lympho penia.Methods One hundred and ten in-patients who were admitted during February 2003 to February 2008 were retrospectively reviewed.Indirect immunofluorescence test was used to detect ALA.Results ① Lymphopenia (＜1.5×109/L) was observed in 68.2% patients.Lymphopenia was associated with skin rashes,serositis,renal involvement,NPSLE,leucopenia,ANA,ds-DNA antibody,ESR and IgG levels.The lymphocyte count and SLEDAI scores was more closely correlated than total white blood cell counts.②ALA was positive in 51/110 (46.4%) patients.ALA was associated with renal involvement,NPSLE,WBC count,C3 level decrease,ANA,dsDNA antibody and SLEDAI scores.Conclusion Lymphopenia is significantly associated with SLEDAI scores.ALA is possibly one cause of lymphcytopenia.In addition,ALA is also a parameter for disease activity,and is associated with organ involvement and outcomes.

OBJECTIVETo investigate the blood supply of the pedicle fat grafts with the third lumbar segmental artery and its clinical effects on reoperation for lumbar disc herniation.

METHODSTwelve sides of 6 adult cadaver examples were contributed to investigate the courser of lumbar segmental vessels and the distribution of hypodermic capillary net of the dorsal branch of the third lumbar segmental artery. From January 2000 to January 2007,49 patients needed reoperation to treat lumbar disc herniation,including 26 males and 23 females with an average age of 55.6 years (ranged from 39 to 70 years). Duration between two operations ranged from 8 months to 15 years with an average of 6.9 years. Reoperative reasons included recurrent lumbar disc protrusion(30 cases)postoperative epidural scar formation (17 cases), postoperative epidural cyst formation (2 cases). Of them,9 patients underwent posterior lumbar interbody fusion at the second operation. The pedicle fat grafts with the third lumbar segmental artery were covered on the sites of the laminectomy in these patients. After negative pressure drainage tube were pulled out, 2 ml Chitsan were injected to the sites of the laminectomy and around epidural nerve root through epidural catheter. VAS score and the Oswestry Disability Index (ODI) were used to assess clinical outcomes before and after operation.

RESULTSThe courser of third lumbar segmental vessels were invariant at the lateral face of the lumbar vertebral body. The dorsal branch of the third lumbar segmental artery penetrated thoracolumbar fascia and formed rich hypodermic capillary net in the region. All patients were followed up from 5 to 8 years with an average of 5.6 years. VAS score of low back pain and leg pain decreased respectively from preoperative 7.6 +/- 1.2, 8.9 +/- 0.9 to 3.6 +/- 0.5, 3.0 +/- 0.4 at final follow-up (P < 0.01); and ODI score decreased from preoperative 44.1 +/- 6.2 to 13.9 +/- 3.6 at final follow-up (P < 0.01). According to ODI score to evaluate the clinical outcomes, 29 cases got excellent results, 11 good, 7 fair, 2 poor.

CONCLUSIONThe pedicle fat grafts with the third lumbar segmental artery and Chitsan can reduce epidural scar formation and prevent peridural fibrosis and adhesion and improve clinical effects of reoperation for lumbar disc herniation.

Adipose Tissue ; pathology ; Adult ; Aged ; Arteries ; pathology ; physiopathology ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; pathology ; physiopathology ; surgery ; Lumbar Vertebrae ; blood supply ; pathology ; surgery ; Male ; Middle Aged ; Reoperation ; Transplantation ; Treatment Outcome

OBJECTIVETo study the effect of all-trans retinoic acid (ATRA) on U937 cell growth and its mechanism.

METHODSCell cycle was detected by flow cytometry (FCM), expressions of cell cycle associated protein and the p27 related protein were detected by Western blot. The binding of P27 and Skp2 was detected by immunoprecipitation.

RESULTSFCM displayed that ATRA could inhibit the proliferation of U937 cells. At 72 h on 1 micromol/L ATRA treatment, 72% of the cells were arrested at G0/G1 phase. Western blot displayed that ATRA could decrease the expression of cyclin A, up-regulate the expression of p21 and p27, and down-regulate the expression of p27 related proteins Skp2. p27 could bind with Skp2 in U937 cells as detected by immunoprecipitation.

CONCLUSIONATRA may arrest the proliferation of U937 cells through the reduction of Skp2 expression, and finally the induction of the accumulation of p27.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; S-Phase Kinase-Associated Proteins ; metabolism ; Tretinoin ; pharmacology ; U937 Cells

OBJECTIVETo study the clinical effects of acromiocoracoid ligament transfer to repair coracoclavicular ligament and acromioclavicular ligament in the treatment of acromioclavicular joint dislocation of type Tossy III in young patients.

METHODSFrom January 2005 to January 2007, 52 patients with acromioclavicular joint dislocation of type Tossy III were divided into therapeutic group I (32 cases) and therapeutic group II (22 cases). There were 17 males and 13 females with an average age of 31.0 years (ranged, 19 to 40 years) in group I; as well as 12 males and 10 females with an average age of 33.6 years (ranged, 20 to 42 years) in group II. All the patients were fresh injury and the acromioclavicular joint dislocations were type Tossy III according to X-ray findings. The duration from damage to the operation time ranged from 2 to 17 days (averaged, 7.6 days). All the patients had normal shoulder function before injury and were treated with clavicular hook plate implantation. The patients in group II were treated with normal desmorrhaphy method, and the patients in group I were treated with acromiocoracoid ligament transfer to repair coracoclavicular ligament and acromioclavicular ligament. Steel plate was taken out at about 4 to 6 months after clavicular hook plate implantation. The results were evaluated according to Karlsson's standards.

RESULTSAll the incisions healed without infection. The therapeutic effects of 52 patients were evaluated at the 6th month after internal fixation was taken out. Twenty-four patients in group I got an excellent results and 6 good; 17 patients in group II got an excellent results and 5 good. There was no significant difference between the two groups. Twenty-six patients in group I and 19 patients in group II had long-term follow-up, and the duration ranged from 3 to 5 years (averaged, 4.5 years ). Twenty patients in group I got an excellent results and 6 good; 9 patients in group II got an excellent results, 7 good and 3 bad. Three patients in group II had recurrence of acromioclavicular joint dislocation. The long-term therapeutic effects of group I was better than that of group II.

CONCLUSIONAfter clavicular hook plate implantation in treating type Tossy III dislocation of the acromioclavicular joint in young patients, it was necessary to use acromiocoracoid ligament transfer to repair coracoclavicular ligament and acromioclavicular ligament, which can improve the stability of acromioclavicular joint to prevent dislocation recurrence.

Acromioclavicular Joint ; injuries ; surgery ; Adult ; Bone Plates ; Case-Control Studies ; Clavicle ; surgery ; Female ; Follow-Up Studies ; Humans ; Joint Dislocations ; surgery ; Ligaments, Articular ; Male ; Treatment Outcome ; Young Adult

OBJECTIVETo study the safety, effectiveness and feasibility of suprapubis-assisted umbilical laparoendoscopic mini-dual-site surgery (SAU-LEMDS) in the treatment of varicocele.

METHODSThis study included 80 varicocele patients aged 24 - 44 (mean 28.5 +/- 2.6) years, 25 cases of grade I, 45 cases of grade II and 10 cases of grade III, 58 cases in the left side, 6 in the right and 16 in both sides, and all with asthenospermia. The patients were treated by SAU-LEMDS under subarachnoid anesthesia combined with general anesthesia in a supine position with a head-down-feet-up slope of 15 degrees. Two 5 mm trocars were inserted bilaterally at the umbilical edge, one with a 5 mm 30 degrees laparoscope placed in it, and another into the abdominal cavity below the pubic hairline with a 5 mm laparoendoscopic clipper placed in it. The operation procedure was similar to that of standard laparoscopic ligation of spermatic veins, with reservation of the spermatic artery and double-ligation of spermatic veins. And the procedure was repeated for the contralateral lesion in the bilateral cases. Postoperative follow-up was conducted for the incidences of orchiatrophy and testicular hydrocele and changes of seminal parameters.

RESULTSAll the operations were successful, with the mean operation time of (10 +/- 5.0) min (range 8 to 25 min) for the unilateral cases and (18 +/- 6.5) min (range 15 to 30 min) for the bilateral cases, the mean blood loss of (1.5 +/- 0.5) ml (range 1 to 2 ml), and the mean postoperative hospital stay of (2 +/- 0.5) d (range 1.5 to 3 d). The patients were followed up for 6 -24 (12 +/- 2.5) months, which showed significant improvement in sperm motility as compared with the baseline ([28.53 +/- 5.21] vs [19.62 +/- 3.56]%, P < 0.05), with 28 cases (35.0%) restored to normal. Recurrence was found in 4 cases (5.0%). Testicular hydrocele occurred in 7 cases (8.75%), but orchiatrophy in none. The scars in the umbilicus and suprapubis were invisible because of the wrinkles and pubic hair.

CONCLUSIONSAU-LEMDS is safe, effective and feasible for the treatment of varicocele. It is superior to umbilical laparoendoscopic single-site surgery (U-LESS) for its less invasiveness, simpler operation, and better cosmetic appearance.

Adult ; Asthenozoospermia ; Humans ; Laparoscopy ; adverse effects ; methods ; Length of Stay ; Ligation ; methods ; Male ; Operative Time ; Postoperative Period ; Recurrence ; Spermatic Cord ; blood supply ; Testicular Hydrocele ; etiology ; Treatment Outcome ; Umbilicus ; Varicocele ; surgery ; Veins

Objective:This study aimed to assess serum fibulin-3 levels in patients with colon cancer compared with healthy controls and its relationship to demographics and tumor pathology.Methods:A total of 97 patients (mean age,59.6 years;57.7％ males) with colon cancer and 86 controls (mean age,58.8 years;54.7％ males) were included.Serum levels of fibulin-3 were determined using a commercially available sandwich ELISA (Enzyme-Linked ImmunoSorbent Assay).Results:Preoperative serum fibulin-3 levels were significantly lower in the group of patients with colon cancer(39.86 ± 24.37) ng/mL compared with the control group (94.25 ± 31.28) ng/mL.Preoperative fibulin-3 levels and negatively correlated with the degree of tumor TNM stage (P＜0.05),fibulin-3 levels gradually decreased by tumor stage from Ⅰ to Ⅳ and the difference is statistically significant.The serum fibulin-3 levels were correlated significantly with tumor size,lymph node metastasis,distant metastasis and CEA levels(P＜0.05).Conclusion:It was concluded that fibulin-3 is expressed at a lower level in colon cancer,and it is significant correlated with tumor burden,decreased level of fibulin-3 can serve as a marker for advanced colon cancer.

Purpose: Nowadays most children with congenital heart disease (CHD) are expected to survive to adulthood. The healthcare focus needs to pay close attention to the important developmental tasks during their growth process. The women with CHD face some challenges in their critically developmental stages during pregnancy, delivery, and even motherhood. The lived experience of being a mother needs to be further concerned. This study aimed to explore the lived experience of first-time mothers with CHD. Methods: Descriptive phenomenological design was adopted. Semi-structured interviews were conducted from April to August 2018 with 11 primiparous women with CHD, who were recruited from the pediatric and adult cardiology outpatient departments at a medical center and who had a child aged between 6 months and 3 years. Giorgi's phenomenological analysis method was employed. Results: Six main themes arose from the analysis: (1) recognizing pregnancy risks, (2) performing self-care for health, (3) building self-worth from my baby, (4) adapting to postpartum life and adjusting priorities, (5) enjoying being a first-time mother, and (6) the factors contributing to success in high-risk childbirth. Conclusions The experiences that occurred prior to and after labor that were identified in this study can assist women with CHD to more capably prepare for and understand the process of becoming a mother, including recognition of the importance of a prepregnancy evaluation. The findings of this study can help women with CHD to better understand the path to becoming a mother and prepare themselves for the challenges that lie ahead.

The study was aimed to analyze the characteristics of LRP16 gene promoter and its activity in order to explore the possible regulation mechanism of LRP16 gene expression. A 2.6 kb genomic DNA sequence of LRP16 5'-end was obtained from NCBI by BLAST software. The 7 target sequences between 0.2 - 2.6 kb from a healthy blood donor DNA sample were amplified by PCR, then identified by DNA sequencing and semi-nest PCR. The verified sequences were analyzed on-line. The results showed that the 7 target sequences were about 400 bp different from each other. All 7 sequences were the same to these GenBank described. At last, all 7 promoter sequences were ligated with luciferase vector, and then the luciferase activity was analyzed in HeLa cells. A known gene promoter sequence can be freely obtained from NCBI database. It is concluded that LRP16 promoter is a standard type II promoter and its activity is strongest in the region from -200 to -600 bp.
Base Sequence ; Chromosomes, Human, Pair 11 ; Gene Expression ; Humans ; Luciferases ; metabolism ; Molecular Sequence Data ; Neoplasm Proteins ; biosynthesis ; genetics ; Promoter Regions, Genetic ; genetics ; Sequence Analysis, DNA

OBJECTIVETo explore the efficacy of tumor-ablative individualized allogeneic hematopoietic stem cell transplantation for the treatment of patients with high risk/refractory leukemia.

METHODSFivety-seven patients with high risk/refractory leukemia were enrolled. Tumor-ablative individualized conditioning regimens included HDAra-C + Bu/Cy, Ara-C + Bu/Fludarabine, G-CSF primed HDAra-C + Bu/Cy, and FLAG followed by reduced-intensified BuCy. Overall survival (OS), disease free survival (DFS), graft versus host disease, infection and relapse post grafting were analyzed.

RESULTSFifty-six patients attained durable engraftment. The median follow-up duration was 17.5 (2 - 34) months. The 18 months probabilities of OS and DFS were (74.7 ± 6.1)% and (62.4 ± 6.7)%, respectively. In addition, the 18 months probabilities of OS and DFS in patients who attained complete remission (CR) before transplantation were (74.2 ± 7.1)% and (58.8 ± 8.1)%, respectively, while in those not attained CR were (77.0 ± 11.8)% and (72.7 ± 11.7)%, respectively. Twenty nine patients developed acute GVHD (aGVHD) (grade I in 18, grade II in 4, grade III in 2 and grade IV in 5). The probabilities of aGVHD was (50.9 ± 6.6)% by Kaplan-Meier curve analysis. The probabilities of grades 2-4 and grades 3-4 aGVHD were (19.3 ± 5.2)% and (12.3 ± 4.3)% respectively. Extensive chronic GVHD (cGVHD) was observed in 36 patients. The probabilities of cGVHD was (64.3 ± 6.4)% by Kaplan-Meier curve analysis. Cytomegaloviremia (CMV) was observed in 39 (68.42%) patients, hemorrhagic cystitis in 13 (22.8%) patients, fungous infection in 16 (28.07%) patients and bacterial infection in 38 (66.67%) patients. Relapse occurred in 14 patients (hematologic relapse in 11 and extramedullary relapse in 3), probabilities of relapse being (24.6 ± 5.7)%. The 17.5-month probability of relapse in patients who attained CR before transplantation was (28.1 ± 7.7)%, while in those not attained CR was (15.6 ± 10.2)%. Fifteen patients died (6 from hematological relapse, 5 from infection of bacterial and fungous, 4 from cGVHD) after 100 days.

CONCLUSIONTumor-ablative individualized allogeneic hematopoietic stem cell transplantation is a promising and safe choice for treatment of high risk/refractory leukemia, even with high leukemia burden.

Cytarabine ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; Transplantation Conditioning