OBJECTIVETo introduce a technique pertaining to S2 iliosacral screw insertion.

METHODSThe screw pathway was first measured on the preoperative pelvic CT scan or the standard sacral lateral radiograph to make sure the existence of the "safe zone" in the S2 segment for screw insertion. Under general anesthesia, patients were positioned supine or prone, depending on the injury pattern of pelvic ring or associated injuries requiring concomitant operation. The operation field was routinely sterilized using iodine and subsequent alcohol solution and draped. The tip of a guide wire was inserted through a stab wound to the posterior outer iliac table, manipulated in the "safe zone" being enclosed by the anterior aspect of the S2 nerve root tunnel, the anterior aspect of the sacral vertebrae, and the inferior aspect of the S1 foramen under the guidance of the standard sacral lateral fluoroscopy, and then the tip was hammered one to two millimeters into the iliac cortex. The guide wire progressed along the trajectory between the inferior aspect of the S1 foramen and the superior aspect of the S2 foramen on the pelvic outlet fluoroscopic view, and then along the posterior to the anterior aspect of the S2 sacral vertebrae and alae on the pelvic inlet fluoroscopic view with a predetermined length. At that moment, in order to ensure the safety, another standard sacral lateral view was imaged to detect the guide wire's tip which should locate posterior to the anterior aspect of the sacral vertebrae and anterior to the anterior aspect of the S2 nerve root tunnel. Subsequently, the depth was measured, the trajectory was drilled and tapped, and the screw was inserted. Following the removal of the guide wire, the wound was irrigated and sutured.

RESULTSUtilizing this insertion technique, there were 30 S2 iliosacral screws in total being placed to stabilize the injured and unstable posterior pelvic ring in 27 patients. Each S2 screw was accompanied by an ipsilateral S1 screw. The S2 screw location was completely intraosseous in all patients, which was verified by postoperative pelvic outlet and inlet radiographs and CT scans. The insertion accuracy was 100 percent in the present series.

CONCLUSIONThe S2 iliosacral screw insertion technique is safe and reproducible to guide the placement of the S2 screw, enhancing the stability for the compromised posterior pelvic ring.

Adult ; Bone Screws ; Female ; Fractures, Bone ; surgery ; Humans ; Ilium ; injuries ; surgery ; Male ; Sacrum ; injuries ; surgery

A new method for establishing ES cell lines from 129/ter. C57BL/6J mice was set up which was characterized by the murine embryonic fibroblast cell(MEF) feeder, the medium of rat heart cell-conditioned medium(RH-CM) for ES cells, and the consecutive digestion by the digestion liquid containing 1% serum. Every group of improved experiments was done with a control of routine method. The results showed that, compared with routine method, the improved way increased the ratio of ES cell lines of 129/ter mice from 11.8% to 33.3%, and of C57BL/6J from 3.7% to 13.3%. The difference is distinct. The passage culture of ES cells showed that, compared with medium added LIF, RH-CM not only inhibited the differentiation of murine ES cells, maintained its dipoild karyotype, but also promote its adherence growth. This kind of culture condition not only maintained the ES cells in an undifferentiated state and their normal dipoild karyotype, but also a series of other characteristics of totipotent embryonic stem cells during extended culture period.
Alkaline Phosphatase ; metabolism ; Amino Acid Sequence ; Animals ; Base Sequence ; Cell Differentiation ; Cell Division ; Cell Line ; Embryo, Mammalian ; cytology ; Female ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Rats ; Stem Cells ; physiology

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism

OBJECTIVETo evaluate the influence of Chinese drugs for cooling blood and dissolving stasis (CBDS) in different concentrations on morphology of krypton laser induced choroidal neovascularization (CNV) in brown Norway (BN) rats.

METHODSForty-eight rats received laser irradiation (659 nm) on fundus of one eye (power 360 mW, spot diameter 50 microm, time 0.05 s). They were divided into four groups equally: the control group (A) treated with normal saline, and the three CBDS groups treated respectively with high (B, 5.0 g/kg), median (C, 2.5 g/kg) and low (D, 1.25 g/kg) dosage of CBDS, twice every day via gastric perfusion for 21 successive days. Fundus fluorescein angiography (FFA) on 4 selected rats in each group was performed at the 7th, 14th and 21st day after photocoagulation, and histopathologic examination using light microscope with immuno-histochemical stain was conducted on them as well.

RESULTSFFA showed that CNV was firstly appeared on day 7 after photocoagulation, in Group A, it expanded gradually and reached the peak on day 21, but showed no significant expansion in the three CBDS groups. The fluorescein leakage in Group C (52343.13 +/- 12973.92 dots) and D (66252.78 +/- 20659.71 dots) was significantly less than that in Group A (91457.19 +/- 29309.11 dots) and B (95973.40 +/- 53950.43 dots) on day 21, all showing statistical significance (P<0.05). The variation of CNV in thickness showed that in Group A it increased gradually from day 7 and reached the peak on day 21 (55.3383 +/- 8.5036 microm); but in the CBDS groups, the peak was reached on day 14, then became thinned gradually, on day 21, it was 40.0913 +/- 13.3448 microm in Group B, 38.8473 +/- 7.9862 microm in Group C and 38.9372 +/- 5.1728 microm in Group D, all thinner than that in Group A significantly (P<0.05).

CONCLUSIONCDBS can effectively suppress the krypton laser induced CNV proliferation and prevent the CNV leakage in BN rats.

Animals ; Choroidal Neovascularization ; drug therapy ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Fluorescein Angiography ; Male ; Rats ; Rats, Inbred BN

Objective To approach the postoperative care method of replantation of amputated finger soaked in fluid.Methods Observation and care of patients' condition were applied to the 25 cases of totally 41 replantation of amputated fingers soaked in fluid from may 2005 to may 2008 after operation.Results Achievement ration was 90% in 25 cases of 41 replantation of amputated fingers.Conclusions Close observation and proper nursing intervention on the operative fingers are the important guarantee to the replantation of amputated fingers.

Objective To investigate the effect of collagen scaffold loaded with collagen-binding domain neurotrophin-3 (CBD-NT3) on the extension of cellular processes of dorsal root ganglions (DRGs), and explore the significance of this kind of combinatorial strategies in the spinal cord injury repair. Methods The tail tendons of SD neonatal rats were performed the removal of cellular components to prepare the collagen scaffold; HE staining was employed to evaluate whether the cells were completely removed from the collagen scaffold. The collagen scaffold was loaded with CBD-NT3,and then, they were co-cultured with primary DRGs for 1, 3 and 5 d, respectively. NT3 and PBS were also co-cultured with primary DRGs for 1, 3 and 5 d, respectively, as controls. Cells on the scaffold were stained by fluorescein diacetate (FDA) for morphology observation and the lengths and angles of the processes in each group were also quantitatively analyzed. Scanning electron microcopy (SEM) was employed to observe the topography of scaffold and the ultrastructure of DRGs 3 d after the co-culture.Results HE staining indicated that the cellular components in the scaffold were removed completely.The length of processes elongation in CBD-NT3 treatment group was significantly longer than that in the controls 3 d after the co-culture (P＜0.05). The 95% confidence interval of the angle between the line which the process emerged from the cell soma to the growing tip of the process and the long axis of fiber was 18.8-20.7 degrees. The results of SEM showed that cells could rely on the topography of the scaffold to anchor and grow. Conclusion The combinatorial strategies of collagen scaffold with CBD-NT3 can play a double function for oriented guiding and inducing extension of cellular processes effectively,which may provide a better therapeutic approach for spinal cord injury repair.

Objective ] To study the molecular characteristics and antibiotic resistance of Salmonella diarrhea cases in sentinel hospitals through active surveillance system conducted by public health laboratory. [ Methods] Two sentinel hospitals were chosen for collection of stool specimens from food-borne infectious diarrhea cases and for Salmonella separation and detection immediately following serotyping and antimicrobial susceptible test ( AST ) on those isolates .Moreover , pulsed field gel electrophoresis ( PFGE) was used for the genetic homology analysis . [ Results] A total of 2 579 diarrhea specimens were collected and analyzed from 2010 to 2012, with 185 Salmonella isolates, covering 23 different serotypes (annual positive rates were 9.1%, 6.8%, 5.1%, with an average 7.2%).Salmonella enterica serovar Enteritidis(S.Enteritidis) and Typhimurium(S.Typhimurium) were the most common serotypes, of which 68.9% cases were seen in those aged 21 to 60 and 21.4% cases in those over 60 years old. 27.7%-96.9%S.Enteritidis and 2.6%-63.2% S.Typhimurium(P all <0.05) proved resistant to Nalidixic acid, Sultisoxazole, Streptomycin, Sulfamethoxydiazine, Gentamicinand Tetracycline. PFGE analysis on 22 S.Enteritidis strains showed 11 different clusters , while 20 S.Typhimurium strians showed 6. [Conclusion] S.Enteritidis and S.Typhimurium are the most common Salmonella serotypes, and molecular typing indicates the existence of clustering and sporadic outbreaks caused by dominance clones . We should be alert to early warnings on potential outbreaks of multiple-drug-resistant ( MDR ) S.Enteritidis.Active surveillance system based on public health laboratory should play an important role in the control of food-borne infectious diseases .

OBJECTIVETo compare the detection sensitivity of epidermal growth factor receptor (EGFR) mutations between allele specific oligonucleotide PCR (ASO-PCR) and bi-loop probe and specific primer quantitative PCR (BPSP-qPCR).

METHODSA total of 96 non-small cell lung cancer specimens were selected from West China Hospital from September 2009 to December 2010. ASO-PCR was developed to detect the presence of classical EGFR mutations. A total 39 available specimens were also tested by BPSP-qPCR.

RESULTSEGFR mutation detection rate was 30.2% (26/96) by ASO-PCR. The mutation rate was higher in female than in male patients [45.5% (20/44) vs. 17.3% (9/52), P = 0.003], non-smokers than smokers [44.1% (26/59) vs. 8.1% (3/37), P < 0.001] and adenocarcinomas than other subtypes of lung cancer [37.0% (27/73) vs. 8.7% (2/23), P = 0.01]. Among mutation negative cases by ASO-PCR, BPSP-qPCR increased the rate of detection of 19-del and L858R mutation by 10.3% (4/39) in adenocarcinomas and non-smoking subset. Overall, the mutation detection rate of BPSP-qPCR was higher than that of ASO-PCR [66.7% (26/39) vs. 41.0% (16/39), P = 0.02].

CONCLUSIONBPSP-qPCR has a better detection sensitivity than that of ASO-PCR.

Adenocarcinoma ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; DNA Mutational Analysis ; Female ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; genetics ; Sensitivity and Specificity ; Sex Factors ; Smoking

OBJECTIVETo study the effects of osteopontin (OPN) on the phenotypes of human hepatocellular carcinoma cell line SMMC-7721.

METHODSHuman hepatocellular carcinoma SMMC-7721 cells were transfected with plasmid pcDNA 3.1(-)/OPN and cells transfected with a mock plasmid served as controls. OPN expression was verified by RT-PCR and Western blot, and concentrations of OPN, MMP-2, MMP-9 and uPA were measured by ELISA. A series of functional assays in vitro were used to monitor the changes of SMMC-7721 malignant phenotypes.

RESULTSOPN expression of SMMC-7721 cells was elevated after transfection. Concentrations of OPN, MMP-2 and uPA in the medium of SMMC-7721 cells after transfection were higher than those of the controls [(3.02+/-0.12) ng/ml vs (1.43+/-0.07) ng/ml, (43.04+/-3.06) ng/ml vs (22.15+/-4.34) ng/ml, and (4.78+/-0.70) ng/ml vs (1.61+/-0.34) ng/ml respectively, P less than 0.01], but MMP-9 concentration did not increase [(7.82+/-2.25) ng/ml vs (7.70+/-1.92) ng/ml]. Functional assays in vitro indicated that SMMC-7721 cells after transfection showed higher adhesive, migrant and invasive capabilities than those of the controls (cell adhesion rates were 75.33%+/-10.59% vs 57.34%+/-2.52%; number of outer surface cells in migrant assay was 14.33+/-2.51 vs 6.34+/-1.53; cell number in the invasive assay was 8.23+/-1.53 vs 4.12+/-1.29 respectively, P less than 0.05).

CONCLUSIONOPN might enhance the expression of MMP-2 and uPA to promote malignant phenotypes of SMMC-7721 cells.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; secretion ; Osteopontin ; genetics ; metabolism ; Transfection ; Urokinase-Type Plasminogen Activator ; secretion

OBJECTIVEEstablishing cDNA microarray, in order to study functional genomics of Salvia miltiorrhiza.

METHODTotal RNA samples were prepared from S. miltiorrhiza roots using a modified CTAB method. mRNA was isolated by Quichprep Micro mRNA Purification Kit from Pharmacia. Then cDNA was synthesized and cloned into the EcoRI-XhoI sites of the ZAP Express vector using a cDNA synthesis kit, and the ligation mixture was packaged using a ZAP-cDNA Gigapack Gold III cloning kit (Stratagene). The single phage was isolated for PCR amplification, Aliquots of the PCR reactions were analyzed in a 1.5% agarose gel to verify the quality of PCR. The remaining cDNA was purified by Multiscreen filter plates (Millipore) and aliquots were analyzed by agarose gel again to verify the quality of purification. Clones passed verification was resuspended in 15 microL 50% DMSO for arraying. An actin gene from S. miltiorrhiza was used for positive control. PloyA and 50% DMSO was used for negative controls.

RESULTBacterial colonies containing cNDAs of S. miltiorrhiza were inserted with average insert size of 0. 5 kb to 2. 5 kb. Total 4 354 genes were singled out from the first 8 736 PCR product and used for cDNA microarray manufacture. Single color fluorescence hybridization showed that all positive controls had signals while negative controls had no signals.

CONCLUSIONIt was the first cDNA microarray about traditional Chinese herbs especially for geoherbs. It could be a powerful tool for studying functional genomics of S. miltiorrhiza.

Gene Expression Profiling ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; methods ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Salvia miltiorrhiza ; genetics