Anthracosis ; complications ; Cerebrovascular Disorders ; complications ; Cohort Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies

In this study ,a wild type Salmonella typhimurium (S .typhimurium) strain was isolated and identified in Hong Kong (S129) ,then the asdA gene was knocked out and replaced with kanamycin resistant gene in a Salmonella typhi-murium strain S129 using the λ RED-mediated recombination method .The constructed mutant asdAΔS129 was validated by culturing in the presence or absence of 2 ,6-diaminopimelic acid (DAP) growth in vitro and evaluating its virulence in BALB/c mice challenge assay .Therefore ,this study has demonstrated that an asdA mutant Salmonella typhimurium has been success-fully constructed .

Objective To preliminarily optimize the acupuncture protocol in treating migraine in acute stage. Method Ninety patients with migraine in acute stage were observed, with Visual Analogue Scale (VAS) as the evaluation index and an orthogonal design. Acupoints groups [Taiyang (EX-HN5), Fengchi (GB20), Taiyang (EX-HN5) and Fengchi (GB20)], insertion directions (perpendicular, downward penetration, and backward penetration), stimulation dosage (1 needle, 3 needles, and 5 needles) and acupuncture duration (30 min, 1 h, and 2 h), altogether 4 factors and 3 levels, formed up different acupuncture protocols to observe the analgesic efficacy in treating migraine in acute stage, so as to determine the role of the four factors (Chi-square test), advantage of the 3 levels (multiple comparisons) and the optimal grouping of the 4 factors and 3 levels. Result Acupoints group, insertion direction and stimulation dosage were the major factors in acupuncture analgesia, and the acupuncture duration was the secondary factor (P<0.05). The analgesic effect of Taiyang (EX-HN5) and Fengchi (GB20) was more significant than either Taiyang (EX-HN5) or Fengchi (GB20) (P<0.05). Penetration puncture from Naokong (GB19) towards Fengchi (GB20) (downward penetration) produced a more significant analgesic effect than from Fengchi (GB20) towards Wangu (GB12) and from Tianyou (TE16) towards Fengchi (GB20) (both backward penetration) (P<0.05). Acupuncture with Five needles and 3 needles (parallel horizontal insertion) were superior to that with 1 needle (P<0.05); acupuncture with 5 needles was better than that with 3 needles but without a statistical significance (P>0.05). Needle retaining for 2 h produced a better analgesic effect than retaining for 30 min and 1 h, but without statistical significances (P>0.05). Conclusion Taiyang (EX-HN5) plus Fengchi (GB20), downward penetration acupuncture, 5-needle parallel horizontal acupuncture, and 2-h needle retaining combine an optimal acupuncture protocol in treating migraine in acute stage. Nevertheless, 3-needle parallel acupuncture with 30-60-min needle retaining can also be chosen according to the condition of the patients.

OBJECTIVETo investigate the prognosis factors of soft tissue sarcoma, especially the impact of surgical treatment on the prognosis.

METHODSWe retrospectively reviewed 208 surgically treated patients. There were 128 male and 80 female. The average age was 46 ranged from 9 to 98 years old. Possible factors of whether the patient firstly treated in our hospital, the tumor size (< 5 cm, 5 ∼ 10 cm, > 10 cm), tumor depth (superficial deep fascia, under the deep fascia), histological type (such as adipose sarcoma, malignant fibrous histiocytoma, synovial sarcoma, fibrous sarcoma, malignant peripheral nerve sheath tumors, other tumors), tumor grade (FNCLCC I, II, III), surgical margin (intralesional, marginal, wide, radical) and adjuvant therapy on the prognosis of patients were analyzed.

RESULTSThe median follow-up was 37.5 ranged from 1.3 to 128.1 months. The overall 3-year and 5-year survival were 77% and 75%. The overall 3-year and 5-year recurrence rate were 28% and 37%. The overall 3-year and 5-year metastasis rate were 35% and 43%. Tumor size, tumor grade and metastasis or not independently affected survival (χ(2) = 18.813, 24.849 and 21.107, all P < 0.05). Whether the patient firstly treated in our hospital and histological type independently affect the local recurrence (χ(2) = 21.915, 12.192, both P < 0.05); histological grade can independently affect the metastasis (χ(2) = 7.714, P < 0.05). Surgical margin alone affected the local recurrence and metastasis (χ(2) = 19.610, 9.272, both P < 0.05).

CONCLUSIONSSurgical margin independently affected local recurrence and distant metastasis, and thus indirectly affect the survival of soft tissue sarcoma. In particular, the primary choice for treatment of soft tissue sarcoma without metastasis should be surgery. Wide or radical margin could significantly improve the prognosis of soft tissue sarcoma patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chi-Square Distribution ; Child ; Extremities ; pathology ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Logistic Models ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Prognosis ; Retrospective Studies ; Sarcoma ; diagnosis ; surgery ; Soft Tissue Neoplasms ; diagnosis ; surgery ; Young Adult

OBJECTIVETo investigate the antitumor effect of recombinant IFN-alpha-2b-BCG on mouse bladder cancer MB49 cells in vitro, and to explore its antitumor mechanisms.

METHODSMB49 cells were co-cultured with recombinant BCG or wild BCG, and than were examined by light and transmission electron microscopy. The cell growth was assessed by MTT assay, and apoptosis rate and MHC-I of the MB49 cells was detected by flow cytometry using AO and Hoechst33258 fluorescence immunostaining.

RESULTSThe hIFN-alpha-2b-BCG-treated tumor cells showed slow growth, detachment of some cells, and various degree of degeneration. Light microscopy revealed organelle disorganization, chromatin aggregation, nuclear pyknosis, and cytolysis in some cells. Cellular membrane bulged and some bubbles were seen under fluorescence microscope using AO staining. Hoechst33258 assay also depicted frequent apoptosis in the tumor cells. The MTT assay showed that rBCG more actively than the wild BCG inhibited the proliferation of MB49 cells. The apoptosis rate of the recombinant BCG group was 19.7% and 46.6% at the time point of 24 h and 48 h, respectively, significantly higher than 10.8% and 20.9%, respectively, in the wild BCG group. The results of flow cytometry indicated that both types of BCG enhanced the expression of MHC-I in the MB49 cells, but more effective in the recombinant BCG group.

CONCLUSIONThe recombinant hIFN-alpha-2b-BCG has more strong immuno-modulatory properties, anti-tumor effect on MB49 cells and induces apparent cytotoxicity in the bladder cancer cells in vitro.

Adjuvants, Immunologic ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; BCG Vaccine ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytotoxicity, Immunologic ; Histocompatibility Antigens Class I ; metabolism ; Interferon-alpha ; pharmacology ; Mice ; Recombinant Proteins ; pharmacology ; Urinary Bladder Neoplasms ; metabolism ; pathology

Background: Wide resection margins of osseous tumors are associated with a low incidence of local recurrence, making accurate measurement of the intraosseous extent of primary malignant long bone tumors is crucial. We compared the intraosseous tumor extent assessed by magnetic resonance imaging (MRI) with the gross specimen to evaluate the accuracy of MRI. Methods: A total of 255 patients with primary malignant tumors in the long bones were included. Using MRI, we defined the length of tumor as the distance from the articular surface to the boundary between abnormal and normal marrow signal. The extent of the abnormal intraosseous signal was measured on unenhanced T1?weighted (T1WI) magnetic resonance images after chemotherapy. All gross surgical specimens were sectioned, and tumor extent was measured. Wilcoxon signed?rank test was used to test the differences between MRI and gross specimen findings. Spearman's correlation analysis was used to test the correlation between groups. Results: Median tumor length by gross specimen (112 mm; range, 45–300 mm) was longer than that by MRI (108 mm; range, 45–304 mm;Z = ?6.916, P < 0.001). Of 255 images, tumor length was accurately represented on 27 T1WI magnetic resonance images, overestimated on 79 images, and underestimated on 149 images. The median difference between imaging and gross specimen measurements was 2.0 mm (range: 1.0–15.0 mm) for the 79 cases where tumor length was overestimated, and 5.0 mm (range: 1.0–18.0 mm) for the 149 cases where tumor length was underestimated. The Spearman correlation demonstrated a high correlation of tumor length on gross specimen with the tumor length on MRI (R = 0.99, P < 0.01). Conclusions: We conclude that preoperative MRI could be a useful method in determining intramedullary malignant bone tumor boundaries and may serve as an accepted assessment method of long bone tumors before limb?sparing surgery.

OBJECTIVETo investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.

METHODSHistological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.

RESULTSCompared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].

CONCLUSIONSAbnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.

Aged ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Female ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Skin ; metabolism ; pathology ; Wound Healing

BACKGROUNDGiant cell tumors (GCTs) most commonly occur around the knee. The most beneficial procedure for this disease has been extensive curettage with reconstruction. However, since many GCTs may compromise the subchondral bone, surgery can further jeopardize the articular cartilage and result in secondary osteoarthritis. In this study, we aimed to determine the factors associated with the development of degenerative arthritis and the effect of bone grafting on the prevention of secondary osteoarthritis.

METHODSWe retrospectively analyzed 76 patients with GCT around the knee. The mean age at first diagnosis was 31.1 years. Surgical treatments included extensive curettage and cementation with or without bone grafting in the subchondral bone. Patient follow-up was a median duration of 35 months, ranging from 18 to 113 months.

RESULTSThe local recurrence rate was 5.3% (4/76). Secondary degenerative changes occurred in 30.3% (23/76) of the patients. Less than 10 mm of the residual thickness of the remaining subchondral bone was correlated with secondary degenerative changes in 57 patients (P < 0.001). Of these 57 patients, 56.5% (13/23) treated with bone cement reconstruction alone developed secondary degenerative changes; following bone grafting, the rate decreased to 29.4% (10/34), with a statistically significant difference (P = 0.041).

CONCLUSIONSGCT patients with less residual thickness of the subchondral bone are more likely to develop degenerative arthritis after curettage. Bone grafting in the subchondral bone area is recommended when the residual thickness of the subchondral bone is less than 10 mm.

Adult ; Aged ; Bone Neoplasms ; pathology ; surgery ; Bone Transplantation ; Female ; Giant Cell Tumor of Bone ; pathology ; surgery ; Humans ; Knee Joint ; pathology ; Male ; Middle Aged ; Retrospective Studies

Objective To observe the expression of vimentin (Vim) after silence of interleukin-1β (IL-1β) in rats with spinal cord contusion (SCC). Methods The model of SCC was established in 30 Sprague-Dawley rats with Allen's method. The rats were randomized into vector group (n=15) and silence group (n=15), which were injected blank lentivirus vector and vector of IL-1β siRNA, respectively; and divided in three, seven and 28 days subgroups. The relationship between IL-1βand Vim was predicted with GeneMANIA bioinformatics. The expression of Vim protein and mRNA in spinal cord was detected with immunohistochemistry and real-time quantitative polymerase chain reaction. Results GeneMANIA bioinformatic analysis indicated that there was some direct and indirect relationship between IL-1β and Vim. The Vim protein and mRNA expressed in the spinal cord, and was less in the silence group than in the vector group (t>2.875, P<0.05). Conclusion Silence of IL-1β can inhibit the expression of Vim in SCC rats, which may promote the recovery of spinal cord function.

Objective To evaluate effect of cytochrome P450 2C19 ( CYP2 C19 ) genotype on pharmacokinetics of voriconazole in healthy volunteers. Methods Studies evaluating the CYP2C19 genetic polymorphism and pharmacokinetics of voriconazole were retrieved through a systematical search of electronic databases ( up to Feb.2015 ).Studies selection according to the inclusion criteria , data extraction and risk of bias assessment were conducted.The meta -analysis was performed using RevMan 5.0.Results Nine studies were included in this meta -analysis, 8 in English, 1 in Chinese.CYP2C19 gene polymorphism had a significantly effect on AUC , poor metabolizers ( PM)>intermediate metabolizers ( HEM ) >extensive metabolizers (EM) >ultrarapid metabolizers (UM), CL(EM>HEM>PM), Cmax ( PM >HEM, EM >UM ) , t1/2 ( PM >EM, PM >HEM ) and tmax ( PM>EM,HEM>EM).Conclusion The pharmacokinetics parameters of voriconazole were influenced significantly by CYP 2C19 genotype in healthy volunteers.