Objective To explore the mutation and evolution of Yersiniapestis(Y. pestis) from the point of codon and 16S-ribosome. Methods Codon preference and 16S-ribosome of Y. pestis were analyzed by bioinformatics. Results Similar codon preference was found among 4 PCD1 Y. pestis, of the 3 old Y. pestis the codon preference between PMT1 and PCD1 was similar. There were some differences between PCD1, PCP1 and Yunnan 6 kb plasmid. Through the analysis of 16S-ribosome, the sequences were found similar in 11 strains of Y. pestis,Yersinia pseudotuberculosis was very close to Y. pestis, with only one nucleotide difference, mutated G-T, and corresponding amino acid methionine (M)-isoleucine (I). There were some differences in sequences of 16S-ribosome in Y. pestis, Escherichia coli and Pulex irritans. Conclusions The time for Y. pestis to obtain PCP1 is later than PMT1 does, in other words, the affinity of Y. pestis with PMT1 was closer than PCP1 with 6 kb plasmid;alteration of 16S-ribosomal nucleotide sites may cause changes in function and structure of 16S rRNA. The lower similarity between 16S-ribosomal sequences of Y. pestis and Pulex irritans indicates the time for co-evolution is very short,and the late emergence of Y. pestis.

Objective To investigate the rat bone structural changes of lower limb following lumbar nerve dorsal roots section.Methods Forty-eight mature female Wistar rats were divided into posterior radi- cotomy(PR)and comtrol groups randomly.The bilateral femoral bone mineral density(BMD)and biome- chanics characteristics were analyzed 2,4 and 8 weeks after the radicotomy.The same operation except the radicotomy was done in the sham group.Results In PR group,2,4,and 8 weeks after the radicotomy,the BMD of femur was(0.221?0.008)g/cm~3,(0.213?0.015)g/cm~3 ,and(0.216?0.105)g/cm~3 ,respective- ly;while that was(0.223?0.005)g/cm~3,(0.218?0.014)g/cm~3 ,and(0.208?0.111)g/cm~3 in control group.No significant difference was observed between the two groups(P＞0.05).In PR group,2,4,and 8 weeks after the operation,the mean maximum load in three-point bending test of femun midshaft was(93.64?8.76)N,(89.77?11.18)N and(93.21?8.74)N,respectively,and was lower than the values of the con- trol group at the same time point(95.94?6.29)N,(91.63?9.43)N,(95.57?8.64)N,However,there was no significant difference between the two groups(P＞0.05).Accordingly,there was no significant difference in the energy absorption in femun midshaft between the two groups(P＞0.05).Conclusion The selective rhizotomies of part lumbar never dorsal roots might not cause the loss of the femur BMD and the change of bio- mechanics property significantly in short period.

OBJECTIVE: To investigate the clinical effect of a modified arthroscopic outside-in suture technique in the treatment of meniscus tear using a spinal needle. METHODS: From January 2015 to October 2017, 95 patients treated with this method were followed-up. Among these cases, there were 36 males and 59 females. The age of the patients ranged from 16 to 77 years, (46.79±18.07) years in average. Among them, there were 28 patieats aged 16-35, 53 patients aged 36-65, and 14 patients aged over 65 years old. 28 cases were diagnosed with medial meniscus tear, 43 cases with lateral meniscus tear and 24 cases with both medial and lateral meniscus tear. Causes of the injury included sports, sprain, etc. According to Barrett standard, the clinical healing of meniscus tear was judged. Lysholm score, knee range of motion, visual simulation score (VAS) and magnetic resonance imaging (MRI) were used to evaluate the postoperative knee function and recovery of the patients. RESULTS: The 95 patients were followed up for 22 to 36 months, with an average of (28.32±3.98) months. According to Barrett standard, 90 patients (94.7%) obtained meniscus clinical healing. Meniscal healing rates were 96.43%, 96.23% and 85.71% in the three age groups, respectively. The meniscal healing rate was lower in the elderly group, but there was no significant difference in statistical results (P=0.262). Five patients had deep tenderness in the joint space of the injured side, and the overstretch test was positive. The preoperative and postoperative VAS scores, Lysholm scores and knee motion were compared in each group, and the differences were statistically significant (P < 0.01). At the end of the last follow-up, there were no cases of knee joint effusion, swelling and interlocking, and the joint function was effectively improved in most patients. No surgical site infection, periarticular vascular/nerve injury or knotting reaction was found during the follow-up. CONCLUSION This modified arthroscopic outside-in suture technique using a spinal needle has the characteristics of simple operation, small trauma and rapid recovery, and the mid-term follow-up results were satisfactory. Therefore, we consider this method to be a safe and efficient method for the treatment of meniscus anterior horn and body tear.
Aged ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Knee Injuries/surgery* ; Male ; Punctures ; Suture Techniques ; Tibial Meniscus Injuries/surgery*

OBJECTIVE: To evaluate the surgical technique and clinical effect of less invasive intrame-dullary nail fixation combined with titanium cable cerclage in the treatment of subtrochanteric fractures. METHODS: A retrospective study was performed in 46 cases of subtrochanteric fractures in Peking University People's Hospital from January 2015 to December 2017. Among them, there were 14 males and 32 females, with an average age of (77.83±10.66) years (44-92 years); 17 cases on the left side and 29 cases on the right side. The causes of injury included crash from a height, traffic accident and accidental fall. According to Seinsheimer classification, there were 26 cases of type Ⅱ, 11 cases of type Ⅲ, 9 cases of type Ⅳ, and these cases were all closed injury. After admission, these patients underwent continuous tibial tuberosity bone traction to maintain the length and force line of the lower extremity, so as to reduce the difficulty of intraoperative fracture reduction. Anticoagulant therapy was given before operation to reduce perioperative thrombotic complications. All the patients were treated with less invasive intramedullary nail fixation combined with titanium cable cerclage. Operation time, blood loss during surgery, time of fracture healing were recorded, Harris and Sanders scoring system were used to assess hip function after operation at each follow-up time point. RESULTS: All the included patients underwent surgery successfully. Average operative time and intraoperative blood loss of these patients were (131.09 ± 20.06) min and (191.96±111.03) mL, respectively. All the patients were followed up satisfactorily, with an average follow-up time of 28 months. The fractures received bone healing within 3-6 months, average hospital stay was (10.61±2.85) days. The Sanders score was excellent in 3 cases, good in 37 cases and common in 6 cases, with an excellent and good rate of 86.96%. The Harris score was excellent in 6 cases, good in 36 cases, with an excellent and good rate of 91.30%. There were no cases of wound infection, loss of reduction, nonunion of fracture or internal fixation failure. Hip pain symptoms were effectively relieved in most patients. CONCLUSION Less invasive intramedullary nail fixation combined with titanium cable cerclage can obtain good alignment and stability of fracture ends, which is an effective method for the treatment of subtrochanteric fractures.
Aged ; Aged, 80 and over ; Bone Nails ; Female ; Fracture Fixation, Intramedullary ; Fracture Healing ; Hip Fractures/surgery* ; Humans ; Male ; Retrospective Studies ; Titanium ; Treatment Outcome

BACKGROUNDSacral nerve injury is a common complication of pelvic or sacral fractures. As the sacral nerve courser within the sacrum and has a complex relationship with the surrounding tissues, different parts of the sacral plexus injury have similar clinical symptoms and signs. Since lack of specific imaging technique in the diagnosis of sacral nerve injury, especially on multi-segment, multi-site, how to determine the preoperative location and extent of the sacral nerve injury accurately becomes a concern of the general orthopaedic and images practitioners. This study was conducted to gain an insight into the overall anatomical features of the sacral nerve (SN) on the same slice in high resolution computed tomography (HRCT) reconstruction and to determine the value of this information for the clinical diagnosis of related diseases.

METHODSFifty healthy volunteers and 30 patients (40 sides) with SN lesions confirmed by surgery were scanned using a 16-slice helical CT scanner (Light Speed, GE, USA). Among the patients, 6 with intervertebral disk hernia (6 sides), 8 with spinal stenosis (12 sides), 11 with pelvic trauma (14 sides), 4 with pelvic malignancies (6 sides), and 1 with sacral vertebral tuberculosis (2 sides). The SN multiplanar reconstruction was performed using a UNIX-based SCD4.1 workstation where the image was set on the same slice. All images were stored in the Digital Imaging and Communications in Medicine format. The display of nerves in different sections was analyzed using a five-graded scale with coordinate curves of each individual score. The overall anatomic features visible on the slice were analyzed and the abnormalities of the lesions were studied.

RESULTSThe image of the same slice clearly revealed the shape, running direction, thickness, tension and adjacent anatomy of the S1-S4 nerves. The rank of display rates in different sections was: outward-rotated oblique sagittal > outward-rotated oblique coronal > oblique coronal plane > coronal > sagittal > transverse section. The S5 nerve was partially displayed from the starting point to the segment around the posterior sacral foramen. The overall anatomy of the triangular sacral plexus was only revealed in the oblique outward-rotated sagittal section, while 100% of its individual rami, as well as two or three of the adjacent rami, were displayed from their starting points to the anterior border of the piriformis. The abnormalities included 39 sides of morphological change (97.5%), 38 sides of compression (95.0%), 35 sides of adhesion (87.5%), 32 sides of displacement (80.0%), 34 sides of shrinkage (85.0%), 6 sides of thickening (15.0%), and 2 sides of abruption (5.0%).

CONCLUSIONSThe 16-slice CT multiplanar reconstruction was able to reveal the overall anatomic features of the SN on the same slice. The section of reconstruction was a crucial factor in determining the display capability of various sacral nerves. This technology was valuable in the diagnosis and management of related diseases.

Adult ; Female ; Humans ; Image Processing, Computer-Assisted ; methods ; Male ; Middle Aged ; Sacrum ; innervation ; Spinal Nerves ; diagnostic imaging ; Tomography, X-Ray Computed ; methods ; Trauma, Nervous System ; diagnostic imaging ; Young Adult

OBJECTIVESTo investigate the intralaboratory reproducibility of immunohistochemistry (IHC) testing for HER2 status in breast cancer, and to evaluate the factors which influence the reproducibility. The concordance between monoclonal antibody CB11 and HercepTest was also assessed.

METHODSHER2 overexpression on paraffin sections from thirty-seven cases of breast invasive ductal carcinoma was evaluated using CB11 and the evaluation procedure had been repeated for five times scored the tests together according to the HercepTest and new American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) grading schemes by 2 experienced pathologists together. Reproducibility rates of the five rounds were assessed using Kappa statistic, and the results from two scoring systems were compared. HercepTest kit was applied to the same cases afterward and the results were compared with CB11.

RESULTSSubstantial intralaboratory reproducibility was achieved among 5 rounds tests. Excluding the influence effect of changing antibody lots, the intralaboratory reproducibility was closed to the perfect threshold (Kappa = 0.7858, HercepTest scheme). The results derived from the two grading schemes had an almost perfect agreement (Kappa = 0.8549). The concordance (positive vs. negative) between CB11 and HercepTest was 83.78%.

CONCLUSIONSLaboratory work with strict supervision and more experience will ensure a reliable testing consistency. Reproducibility analysis could be adopted to evaluate the intralaboratory staining quality on HER2 testing. Different antibody lots bring some influence to the intralaboratory reproducibility, but not significant. CB11 could be accepted to screen HER2 status in routine practice after testing validation.

Antibodies, Monoclonal ; metabolism ; Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Female ; Humans ; Immunohistochemistry ; methods ; Receptor, ErbB-2 ; metabolism ; Reproducibility of Results

OBJECTIVETo investigate the effect(s) of interleukin-1beta (IL-1beta) on the activity of human growth hormone (hGH) gene promoter in rat pituitary GH3 cells and the molecular mechanism.

METHODSThe method of luciferase reporter gene was used. We firstly established stable GH3 cell line which contains hGH gene promoter -484-30 bp and luciferase reporter gene. After treating these cells with IL-1beta or IL-1beta plus various signaling transduction inhibitors, the concentration of GH in the medium and lysate of GH3 cells and luciferase activities in GH3 cells were measured to reflect the effect of IL-1beta on secretion and synthesis of GH and the promoter activity of the hGH gene and the molecular mechanism. Results IL-1beta (10-10(4)U/ml) increased secretion and synthesis of GH. IL-1beta at levels of 10(2)-10(4) U/ml promoted the luciferase expression in stable GH3 cells, and the maximal action was 1.61 times of the control (P < 0.001). Among the inhibitors of intracellular signaling transduction pathways, mitogen-activated protein kinases (MAPK) inhibitor PD98059 (40 micromol/L) and p38 MAPK inhibitor SB203580 (5 micromol/L) completely blocked the stimulatory effect of IL-1beta, and phosphoinositide 3-kinase (PI3-K) inhibitor LY294002 (10 micromol/L) partly blocked the induction of IL-1beta. Neither overexpression of Pit-1 nor inhibiting Pit-1 expression affected IL-1beta induction of hGH promoter activity. The stimulatory effect of IL-1beta was abolished following deletion of the -196 to -132 bp fragment.

CONCLUSIONSIL-1beta increases the activity of hGH gene promoter in rat pituitary GH3 cells. This stimulatory effect of IL-1beta appears to require the intracellular MAPK, p38 MAPK, and PI3-K dependent signaling pathways. The effect of IL-1beta requires the promoter sequence that spans the -196 to -132 bp fragment of the gene, but it is unrelated to Pit-1 protein.

Animals ; Cell Line ; Chromones ; pharmacology ; Flavonoids ; pharmacology ; Genes, Reporter ; Growth Hormone ; biosynthesis ; genetics ; Humans ; Interleukin-1 ; pharmacology ; Luciferases ; metabolism ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Pituitary Gland ; cytology ; enzymology ; metabolism ; Rats ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases

Objective To observe the influences of Ganfujian Formula on the expressions of transforming growth factor-β1 (TGF-β1), TβRII-mRNA, Smad2/3, Smad4 and Smad7 in rats with hepatic fibrosis induced by porcine serum.Methods The hepatic fibrosis model was established in SD rats by the intraperitoneal injection of porcine serum (0.5 mL, twice a week for 8 weeks). The rats then were given Ganfujian Formula orally in different doses (1.50 g/kg, 2.25 g/kg and 3.0 g/kg) and colchicines (0.154 g/kg) respectively until the end of 12th week. The expressions of TGF-β1 and TβRII-mRNA were detected by using RT-PCR and the expressions of Smad2/3, Smad4 and Smad7 were detected by applying Western blotting method.Results In the model group, compared with the normal control group, the expressions of TGF-β1 and TβRII-mRNA, Smad2/3 and Smad4 increased significantly and Smad7 expression decreased significantly. After the treatment with Ganfujian Formula the expressions of TGF-β1 and TβRII-mRNA were reduced in the treatment group compared with the model group. Ganfujian Formula regulated downward the expressions of Smad2/3 and Smad4, and regulated upward Smad7 expression at the same time.Conclusion Ganfujian Formula can inhibit the expressions of TGF-β1 and TβRII-mRNA, regulates downward the expressions of Smad2/3 and Smad4, and regulates upward Smad7 expression. It may be its one of the mechanisms for curing hepatic fibrosis that it has an obvious influence on hepatic TGF-β and Smad signal pathway.

OBJECTIVETo investigate the characteristics of work needed to release adhesion in the early stage of flexor tendon healing.

METHODSEighty-four profoundus flexor tendons of the 3rd toe of Sanhuang chicken were severed and repaired by either Modified Kessler technique or Tsuge technique randomly. At 0, 1, 4, 7, 10, 14, 21 d after the operation, 6 tendons from each repair technique group were harvested and tested by biomechanical test machine. An elongation force-elongation distance curve was obtained and work of adhesion release was calculated in this curve. Whether adhesion band had been released or not was confirmed by gross inspection immediately after the test.

RESULTSWork of adhesion release: this work significantly increased at the 7th day in Modified Kessler group and at the 10th day in Tsuge group. Modified Kessler group was significantly higher than Tsuge group at the 7th day. At the 21st day after operation, adhesion could not be effectively released by full range excursion of flexor tendon profoundus.

CONCLUSIONWork of adhesion release begins to increase significantly since the 7th approximately 10th day postoperatively. At the 21st day after the operation, adhesion could not be fully released by full range of flexor tendon profoundus flexion.

Animals ; Chickens ; Male ; Stress, Mechanical ; Suture Techniques ; Tendon Injuries ; physiopathology ; surgery ; Tendons ; pathology ; physiopathology ; Tensile Strength ; Time Factors ; Tissue Adhesions ; physiopathology ; Wound Healing

BACKGROUNDCalcitonin gene-related peptide (CGRP), a sensory neuropeptide, affects osteoblast proliferation and bone formation. However, the mechanisms are not fully understood. Monocyte chemoattractant protein-1 (MCP-1) is a chemokine that stimulates the migration of monocytes and plays important roles in regulating bone remolding during fracture repair. In this study, we investigated the effects of CGRP on proliferation and MCP-1 expression in cultured rat osteoblasts.

METHODSPrimary rat osteoblasts were isolated from fetal rats calvariae. Cells were exposed to gradient concentrations (10(-9) to 10(-7) mol/L) of CGRP. Protein and mRNA levels of MCP-1 were quantified by Western blotting and semiquantitative reverse transcription-polymerase chain reaction, respectively. The protein level of MCP-1 was investigated and compared in cell culture media by enzyme linked immunosorbent assay (ELISA). Phospho-extracellular signal-regulated kinase (ERK) expression was detected by Western blotting. Cell proliferative activity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and BrdU assay. The effects of MAPK/ERK kinase (MEK)-inhibitor U0126 on CGRP-induced MCP-1 expression in primary rat osteoblasts were examined.

RESULTSCGRP effectively enhanced primary rat osteoblast proliferation and led to significant increases in the expression of MCP-1 mRNA and protein in time- and dose-dependent manners. CGRP activated the ERK pathway. Pretreatment of cultured rat osteoblasts with MEK inhibitor U0126 resulted in dose-dependent inhibitions of CGRP-induced MCP-1 mRNA and protein levels. Thus, CGRP promoted cell proliferation and stimulated MCP-1 expression in cultured rat osteoblasts.

CONCLUSIONThese studies document novel links between CGRP and MCP-1 and illuminate the effects of CGRP in regulating bone remodeling.

Animals ; Blotting, Western ; Butadienes ; pharmacology ; Calcitonin Gene-Related Peptide ; pharmacology ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Chemokine CCL2 ; genetics ; metabolism ; Enzyme Inhibitors ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; metabolism ; Nitriles ; pharmacology ; Osteoblasts ; drug effects ; metabolism ; Rats