Objective To investigate the regulatory role of programmed death ligand-1 (PD-L1) on acute lung injury (ALI), and its molecular mechanism. Methods Twenty C57BL/6 male mice and 20 PD-L1 knock out male mice were collected, and they were divided into two groups by random number table, respectively: namely sham group and ALI group, 10 mice in each group. The model of ALI was reproduced by two-hit of hemorrhagic shock and sepsis, and the mice in sham group were only got bilateral femoral artery exposure and ligation without bleeding, cecal separation without ligation and perforation. The mice were sacrificed 24 hours after model reproduction, and the lung tissue and bronchoalveolar lavage fluid (BALF) were collected. The mRNA and protein expression levels of PD-L1 in the lungs were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western Blot. The pathological changes were observed with microscopy. The protein levels in BALF were determined. The granulocyte differentiation antigen 1 (Gr1) positive cells was determined by cytometry, and myeloperoxidase (MPO) activity in lung tissue was determined. The levels of proinflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and chemotatic factors keratinocyte chemoattractant (KC), macrophage inflammatory protein-2 (MIP-2) in lung homogenates and BALF were determined by enzyme-linked immunosorbent assay (ELISA). Results Compared with sham group, the mRNA and protein levels of PD-L1 in lung tissue of C57BL/6 mice in ALI group were significantly elevated [PD-L1 mRNA (2-ΔΔCt): 3.20±0.76 vs. 1.01±0.03, PD-L1 protein (A value): 0.98±0.16 vs. 0.15±0.04, both P < 0.05]. It was shown by light microscopy that the alveolar wall was thickened, congestive, edema and spot bleeding with a large number of inflammatory cell infiltration in the lung tissue of C57BL/6 mice in ALI group, and an obvious protein leakage was found in BALF (ng/L: 0.18±0.06 vs. 0.05±0.01, P < 0.05). The lung injury degree of PD-L1 knockout ALI mice was significantly less than that of C57BL/6 ALI mice, and the protein leakage was significantly reduced in BALF (ng/L: 0.11±0.02 vs. 0.18±0.06, P < 0.05). Compared with corresponding sham group, the number of Gr1 positive cells, MPO activity in lung tissue as well as the levels proinflammatory factors and chemotatic factors in lung tissue and BALF in ALI group were significantly increased. However, when compared with C57BL/6 ALI mice, above parameters in lung homogenates and BALF were significantly decreased in PD-L1 knockout ALI mice [number of Gr1 positive cells: (39.0±4.0)% vs. (45.0±8.0)%, MPO activity (U·μg-1·min-1): 2.85±0.62 vs. 4.52±1.16; lung IL-6 (ng/g): 461±111 vs. 728±28, TNF-α (ng/g): 1 123±175 vs. 1 500±327, KC (ng/g): 150±34 vs. 250±28, MIP-2 (ng/g): 1 263±468 vs. 1 763±323; BALF IL-6 (ng/L): 134±22 vs. 258±38, TNF-α (ng/L): 598±102 vs. 889±139, KC (ng/L): 934±286 vs. 1 258±336, MIP-2 (ng/L): 650±130 vs. 950±256; all P < 0.05]. Conclusion PD-L1 may play an important protective role in the immunological mechanism of ALI, which may be mediated by decreasing chemotactic factor KC and MIP-2 and mitigating neutrophil chemotaxis in lung tissue.

OBJECTIVETo investigate and research the appraisal scores of the symptoms and physical signs index for the evaluation of the clinical efficacy of acupuncture and manipulative regulation of knee muscle balance for the treatment of knee osteoarthritis, and provide clinical basis for the treatment.

METHODSFrom January 2008 to December 2009, 121 patients with knee osteoarthritis were randomly divided into two groups. In the treatment group there were 63 cases of 83 knees including 16 males and 47 females, with an average age of (59.88 +/- 7.97) years; in the control group there were 58 cases of 73 knees including 13 males and 45 females, with an average age of (57.95 +/- 10.37) years. The patients in the treatment group were treated with acupuncture plus manipulative regulation of knee muscle balance, and the patients in the control group were treated with Diclofenac Sodium Sustained Release Tablets. The appraisal scores of the symptoms and physical signs index, numerical rating scale of pain, joint function, joint swelling were evaluated before and after the treatment, as well as 3 months after the treatment. All data were statistical analyzed by package SPSS 10.0.

RESULTS1) In the treatment group, before and after treatment the appraisal scores of the symptoms and physical signs index were (39.81 +/- 3.92) and (9.69 +/- 8.08); numerical rating scale of pain were (7.61 +/- 0.97) and (2.17 +/- 2.09); joint function were (1.47 +/- 0.50) and (0.61 +/- 0.58); joint swelling were (1.23 +/- 0.79) and (0.42 +/- 0.52). As well in the control group, above data were (39.89 +/- 3.78), (13.62 +/- 7.83), (7.55 +/- 0.71), (3.34 +/- 2.32), (1.33 +/- 0.47), (0.93 +/- 0.67), an (0.97 +/- 0.88), (0.58 +/- 0.52) respectively. Both group had obvious differences in the appraisal scores of the symptoms and physical signs index, numerical rating scale of pain, joint function, joint swelling between after and before treatment. The comparison between the two groups suggested that after the treatments the treatment group had better therapy effect than those of the control group in the appraisal scores of the symptoms and physical signs index, numerical rating scale of pain, joint function, but both group had no differences in the appraisal scores of joint swelling. 2) After the treatment, 12 patients in the treatment group were clinically cured, 48 had remarkable effects,19 good, and 4 ineffective. As well in the control group, above data were 5, 31, 34 and 3 respectively. Three months after the treatment, above data were 8, 42, 27 and 6 respectively in the treatment group. In the control group, above data were 5, 21, 37 and 10 respectively. The treatment group was superior to the control group after the treatments and 3 months after the treatment.

CONCLUSIONAcupuncture plus manipulative regulation of knee muscle balance can effectively improve the clinical symptoms and knee joint's motor function of patients with knee osteoarthritis, and can avoid the further development of disease. It is a proved effective method for knee osteoarthritis.

Acupuncture Therapy ; Aged ; Combined Modality Therapy ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; therapy

OBJECTIVEBone-marrow derived mesenchymal stem cells (BMSC) have the potential to differentiate into endothelial cells. The aim of the study was to investigate the induction process of BMSC by B16 melanoma cells in vitro and to analyze the role of VEGF-a in the process.

METHODSA co-culture system containing BMSC and B16 melanoma cells based on transwell indirect model was established, and the induction process of BMSC by B16 melanoma cells was studied in vitro.

RESULTSBMSC were isolated from the bone marrow of C57 mice. BMSC expressed CD105, CD90, CD73, CD44 and CD166, and acquired expressin of endothelial phenotype markers including VEGFR-1, VEGFR-2 and Factor VIII after co-culture with B16 melanoma cells for 48 hours. The expression level of VEGFR-2 would be double and Factor VIII threefold more by extending the co-culture time to 72 hours. In the co-culture system, B16 melanoma cells also up-regulated the expression of VEGF-a.

CONCLUSIONSVEGF-a plays a significant role in the differentiation of BMSC into cells of endothelial phenotype, therefore, is important to tumor angiogenesis.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Factor VIII ; metabolism ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Pilot Projects ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

To investigate the relationship between HLA-DQB1* alleles and major beta-thalassemia, the HLA-DQB1* loci typing was performed with polymerase chain reaction-sequence specific primer (PCR-SSP) in 42 unrelated (unconsanguineous) patients with major beta-thalassemia and 45 normal control individuals in Guangdong Province. Results showed that the frequency of HLA-DQB1*06 allele in patient group (19.0%) was higher than that in the control group (4.4%) kappa(2) = 8.961, p < 0.01). Our data suggests that HLA-DQB1*06 allele is associated with pathogenesis of the major beta-thalassemia in Guangdong area.
Alleles ; Asian Continental Ancestry Group ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-DQ Antigens ; genetics ; HLA-DQ beta-Chains ; Humans ; Male ; beta-Thalassemia ; genetics

Objective Analyze the relationship between the expression of hypoxia-inducible factor-1α( HIF-1α)and epidermal growth factor receptor(EGFR) and CD105 micro vessel density (MVD) and their value in evaluating biologic behavior and prognosis in laryngeal cancer. Methods Ninty-one cases of laryngeal cancer were analyzed about their clinical and pathology data. In tumor tissue the expression of HIF-1α and EGFR was detected by immunohistochemistry and MVD was marked by CD105. Results The expression of HIF-1α was correlated with size, TNM stage, T stage, lymph node metastasis and histological grade( all P < 0. 05). The expression of EGFR was correlated with TNM stage, lymph node metastasis, histological grade and relapse(all P <0. 05). MVD was correlated with type, TNM stage, T stage, lymph node metastasis, metastasis and histological grade ( all P < 0. 05 ). The expression of HIF-1α and EGFR was correlated with MVD ( F value was 7. 644 and 5. 197 respectively, P value was 0. 001and 0. 025 respectively). The correlation between the expression of HIF-1α and EGFR was significant statistically( r = 0.238,P= 0.007). The survival rate of patients of 3 years and 5 years were 56.1% and 44.2% respectively. Survival analysis by Log Rank showed that prognosis of laryngeal cancer patients was correlated with type, TNM stage and the expression of both HIF-1αand EGFR. While Cox multiple factors analysis demonstrated that TNM stage and expression of EGFR were independent prognostic factor of laryngeal cancer. (P value was 0. 049 and 0. 041 respectively,RR was 1. 300 and 2. 417 respectively). Conclusions HIF-1α and EGFR are key molecular event during development and progression of laryngeal cancer, which act in regulating tumor angiogenesis as well, and show intimate relationship with biological behavior and prognosis of laryngeal cancer.

BACKGROUNDMutation and expression change of p21WAF1/CIP1 may play a role in the growth of osteosarcoma. This study was to investigate the expression of the p21WAF1/CIP1 gene in human osteosarcoma, p21WAF1/CIP1 gene DNA sequence change and their relationships with the phenotype and clinical prognosis.

METHODSp21WAF1/CIP1 gene in 10 normal people and the tumours of 45 osteosarcoma patients were examined using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) with silver staining. The PCR product with an abnormal strand was sequenced directly. The p21WAF1/CIP1 gene mRNA and P21 protein of 45 cases of osteosarcoma were investigated by using in situ hybridization and immunohistochemistry, respectively.

RESULTSThe occurrence of P21 protein in osteosarcoma was 17.78% (8/45), and p21WAF1/CIP1 mRNA expression in osteosarcoma was 42.22% (19/45). The p21WAF1/CIP1 gene DNA sequencing of amplified production showed that in p21WAF1/CIP1 gene exon 3 of 36 cases of human osteosarcoma, there were 17 cases (47.22%) with C-->T at position 609; 10 normal blood samples' DNA sequence analysis yielded 8 cases (80.00%) with C-->T at the same position.

CONCLUSIONSAlong with the increase of malignancy, the expression of p21WAF1/CIP1mRNA and P21 protein in osteosarcoma tends to decrease. It is uncommon for the p21WAF1/CIP1 gene mutation to occur in human osteosarcoma. As a result, the possible existence of tumour subtypes of p21WAF1/CIP1 gene mutation should be investigated. Our research leads to the location of p21WAF1/CIP1 gene polymorphism of Chinese osteosarcoma patients, which can provide a basis for further research.

Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; genetics ; Humans ; Osteosarcoma ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; RNA, Messenger ; analysis ; Sequence Analysis, DNA

BACKGROUNDOld pelvis fractures are among the most challenging fractures to treat because of their complex anatomy, difficult-to-access surgical sites, and the relatively low incidence of such cases. Proper evaluation and surgical planning are necessary to achieve the pelvic ring symmetry and stable fixation of the fracture. The goal of this study was to assess the use of three-dimensional (3D) printing techniques for surgical management of old pelvic fractures.

METHODSFirst, 16 dried human cadaveric pelvises were used to confirm the anatomical accuracy of the 3D models printed based on radiographic data. Next, nine clinical cases between January 2009 and April 2013 were used to evaluate the surgical reconstruction based on the 3D printed models. The pelvic injuries were all type C, and the average time from injury to reconstruction was 11 weeks (range: 8-17 weeks). The workflow consisted of: (1) Printing patient-specific bone models based on preoperative computed tomography (CT) scans, (2) virtual fracture reduction using the printed 3D anatomic template, (3) virtual fracture fixation using Kirschner wires, and (4) preoperatively measuring the osteotomy and implant position relative to landmarks using the virtually defined deformation. These models aided communication between surgical team members during the procedure. This technique was validated by comparing the preoperative planning to the intraoperative procedure.

RESULTSThe accuracy of the 3D printed models was within specification. Production of a model from standard CT DICOM data took 7 hours (range: 6-9 hours). Preoperative planning using the 3D printed models was feasible in all cases. Good correlation was found between the preoperative planning and postoperative follow-up X-ray in all nine cases. The patients were followed for 3-29 months (median: 5 months). The fracture healing time was 9-17 weeks (mean: 10 weeks). No delayed incision healing, wound infection, or nonunions occurred. The results were excellent in two cases, good in five, and poor in two based on the Majeed score.

CONCLUSIONSThe 3D printing planning technique for pelvic surgery was successfully integrated into a clinical workflow to improve patient-specific preoperative planning by providing a visual and haptic model of the injury and allowing patient-specific adaptation of each osteosynthesis implant to the virtually reduced pelvis.

Adolescent ; Adult ; Female ; Fractures, Bone ; diagnosis ; pathology ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Pelvic Bones ; surgery ; Reconstructive Surgical Procedures ; Young Adult

OBJECTIVETo investigate the effect of Bushengzhuyang Fang (Yangjing Capsule, YJC) on penile erectile function and its action mechanisms in rats.

METHODSFifty-six male SD rats were randomly divided into seven groups of equal number: blank control, daidzein, daidzein + testosterone, daidzein + sildenafil, daidzein + low-dose YJC, daidzein + medium-dose YJC, and daidzein + high-dose YJC. The rats in the blank control group were treated intragastrically with normal saline and those in the other groups with daidzein at the dose of 100 mg per kg per day for 30 days. Then the last five groups received additionally testosterone (4 mg per kg per day), sildenafil (2.5 mg per kg per day), low-dose YJC, (0.315 mg per kg per day), medium-dose YJC (0.63 mg per kg per day), and high-dose YJC (1. 26 mg per kg per day), respectively. At 0, 30 and 60 days of treatment, we observed the apomorphine-induced spontaneous erectile response and pathological changes in the corpus cavernosum of the rats, recorded the number of penile erection and erectile incubation period, and determined the serum levels of testosterone (T) and luteinizing hormone (LH).

RESULTSAt 30 days of treatment, the number of apomorphine-induced erections was decreased, the erectile incubation period prolonged, and the serum levels of T and LH reduced remarkably in all groups of rats (P < 0.05). Compared with the findings at 30 days, the number of penile erections was significantly decreased at 60 days in the daidzein group (1.39 ± 0.42 vs 2.67 ± 0.33, P < 0.05) and daidzein + low-dose YJC group (1.33 ± 0.49 vs 2.83 ± 0.61, P < 0.05); the erectile incubation period was markedly ex- tended ([16.33 ± 3.11] vs [8.50 ± 0.93] min and [15.50 ± 3.21] vs [8.63 ± 1.54] min, P < 0.05); and the serum levels of T ([5.34 ± 0.89] vs [1.24 ± 0.30] ng/ml and [5.28 ± 1.12] vs [2.07 ± 0.76] ng/ml, P < 0.05) and LH ([3.62 ± 0.37] vs [2.09 ± 0.12] ng/ml and [3.79 ± 0.28] vs [2.17 ± 0.33] ng/ml, P < 0.05) were significantly reduced in the daidzein and daidzein + low-dose YJC groups, respectively. Pathological examination revealed slightly decreased cavernous sinuses and blood vessels in the corpus cavernosum of the rats in the daidzein + testosterone, daidzein + sildenafil, daidzein + medium-dose YJC, and daidzein + high-dose YJC groups as compared with those in the blank control group.

CONCLUSIONHigh-dose Yangjing Capsule is efficacious for the recovery of erectile function in rats, especially for phytoestrogen-induced erectile dysfunction.

Animals ; Apomorphine ; pharmacology ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; chemically induced ; drug therapy ; Humans ; Isoflavones ; pharmacology ; Luteinizing Hormone ; Male ; Penile Erection ; drug effects ; physiology ; Penis ; drug effects ; pathology ; Phytoestrogens ; Phytotherapy ; Piperazines ; therapeutic use ; Purines ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Sildenafil Citrate ; Sulfonamides ; therapeutic use ; Testosterone ; therapeutic use ; Vasodilator Agents ; therapeutic use

OBJECTIVETo investigate the effect of interstitial cells of Cajal (ICC) on contraction of intestinal tract smooth muscle induced by motilin receptor agonist.

METHODSTwo kinds of smooth muscle segments were isolated from the duodenum and colon of rabbit. Both kinds of smooth muscle were divided into two groups: group a (normal ICC group of duodenum); group c (impaired ICC group of duodenum); group b (normal ICC group of colon); group d (impaired ICC group of colon), each group contained 20 segments. The impairment of ICC was induced by selectively destroying ICC in the smooth muscle via treatment with methylene blue plus light. Then the frequency and amplitude of contraction of group a and c, group b and d was compared. Then motilin receptor agonist (ABT-229) was added into the Krebs solution, the frequency and amplitude of smooth muscle contraction before and after adding ABT-229 were recorded and compared.

RESULTSThe electron microscopy demonstrated that ICC in methylene blue plus light group were destroyed; the smooth muscle cells and neuron scattered close to ICC were normal. In group a, the contraction frequency, (17.89 +/- 1.88) times/min, was significantly lower as compared with that measured after ABT-229 was added [(18.76 +/- 1.18) times/min (P > 0.05)]; the amplitude of group a was (343 +/- 28) mg, which was lower as compared with that after adding ABT-229 [(597 +/- 68) mg (P < 0.001)]; in group b, the frequency was (5.89 +/- 1.03) times/min, the amplitude was (724 +/- 85) mg, after ABT-229 was added, the construction frequency increased to (8.45 +/- 0.69) times/min (P < 0.001), and the amplitude was (897 +/- 89) mg (P < 0.05), which was not affected by pretreatment with TTX, however it could be weakened by nifedipine significantly. In group c and d, the rhythmic contraction almost disappeared: in group c the contraction frequency was (1.06 +/- 0.24) times/min, and the amplitude were (50 +/- 10) mg. In group d, the amplitude and frequency significantly decreased as compared with the normal group (P < 0.001), in group c, and d, no significant difference in amplitude and frequency was found between the values measured before and after adding ABT-229 (P > 0.05). After Ach (100 micromol/L) was added, both group c and d could generate contraction.

CONCLUSIONICC may play an important role in the rhythmic contraction of intestinal tract. The promoting effect of motilin receptor agonist on intestinal tract may be mediated by ICC. ICC deficiency may cause functional impairment of gastrointestinal tract motivation. The medication may become ineffective when the number of ICC is reduced to a certain extent or the network of ICC is incomplete.

Animals ; Erythromycin ; analogs & derivatives ; pharmacology ; Female ; Gastrointestinal Motility ; drug effects ; physiology ; Interstitial Cells of Cajal ; physiology ; Male ; Rabbits ; Receptors, Gastrointestinal Hormone ; agonists ; Receptors, Neuropeptide ; agonists

OBJECTIVETo localize susceptibility loci in Chinese systemic lupus erythematosus (SLE) cohort by linkage disequilibrium mapping the genomic interval in human chromosome 16 so as to understand whether the pathogenesis of human SLE is related to chromosome 16.

METHODSFive microsatellite markers in chromosome 16 spanning from 57.79 cM to 65.1 cM were used for fluorescence based genotyping in 157 SLE families. Evidence for linkage disequilibrium was assessed using the extended transmission disequilibrium test (ETDT) program and Genehunter software. Susceptibility gene was searched in the positive locus, and real-time PCR method was used to detect gene expression.

RESULTSWith the ETDT, evidence for linkage disequilibrium of the marker D16S409 (P=0.0278) and D16S517 (P<0.0001) with SLE were found. It was observed that 271 bp allele of D16S517 was much in evidence for preferential transmission, while 277 bp allele was found to be transmitted less often than expected. Genehunter analysis is concordant with ETDT. By real-time PCR, OAZ(OLF1/ EBF-associated zinc finger protein) gene which is located in the positive locus showed higher expression in SLE patients than in normal controls.

CONCLUSIONResults reveal that 16q12 (58.46 cM) is associated with Chinese SLE. OAZ is a likely susceptibility gene in this locus for SLE.

Adolescent ; Adult ; Alleles ; China ; Chromosomes, Human, Pair 16 ; genetics ; Cohort Studies ; DNA ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Microsatellite Repeats ; Middle Aged