According to Chinese medicine, the atlantoaxial joint is a composite joint composed of tendons and bones, and the stability of the joint depends on the 'tendon-bone balance' involving tendons, ligaments, atlas and axis. Multiple causes of 'tendon off-position, joint subluxation' will lead to joint 'tendon-bone imbalance', which will evolve into atlantoaxial subluxation (AAS), endangering human health. Chinese therapeutic massage (tuina) is a very effective treatment for AAS in adults, but conventional manipulations are prone to ineffectiveness or accidents due to neglect of the causal relationship of the 'tendon-bone imbalance' and inappropriate manipulations. Compared with conventional manipulations, the rational choice of modified manipulations under the guidance of 'tendon-bone balance' theory is more effective and less risky, and more worthy of clinical promotion. From the 'tendon-bone balance' theory, we considered the shortcomings of conventional manipulations, and introduced several modified manipulations that have their own strengths in 'tendon smoothing' and 'bone setting', in order to provide new ideas for treatment of AAS in adults.

This study was aimed to construct model cell line of NPM1-RNAi in HL-60 cells and its resistant line (HL-60/ADR) so as to provide a experimental basis for investigating the potential role of NPM1 gene in leukemia drug resistance. The shRNA targeting to NPM1 was ligated into linear pGCSIL-GFP vector, and transformed into E.coli DH5α. Positive clone was identified by PCR and DNA sequencing. pHelper 1.0, pHelper 2.0 and pGCSIL-GFP-NPM1-shRNA were cotransformed into 293T cells by lentivirus vector system. NPM1-RNAi-LV was transfected into HL-60 and HL-60/ADR cell lines. The efficiency of NPM-RNAi-LV was detected by using real-time quantitative RT-PCR and Western blot. The results showed that the recombinant eukaryotic expression vector pGCSIL-GFP-NPM1-shRNA was constructed. pGCSIL-GFP-NPM1-shRNA was packed into NPM1-RNAi-LV by lentivirus vector system, and transfected into HL-60 and HL-60/ADR cell lines. At mRNA level, the efficiency of NPM1 mRNA knockdown was more than 90% (p < 0.05). At protein level, obvious down-regulation of NPM protein was noted, indicating that NPM1 gene in HL-60 and HL-60/ADR cell lines was knocked down after transfected with NPM1-RNAi-LV. The resistance of HL-60/ADR cell line to adriamycin decreased to a certain degree after NPM1 gene silencing. It is concluded that the model cell lines of NPM1-RNAi in HL-60 and HL-60/ADR are successfully constructed, which can be used for investigating the potential role of NPM1 gene in drug resistance of leukemia.
Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; HL-60 Cells ; Humans ; Lentivirus ; genetics ; Nuclear Proteins ; genetics ; RNA Interference

Objective:To investigate the computed tomography (CT) features of gastric neuroendocrine neoplasm (GNEN).Methods:The retrospective and descriptive method was conducted. The clinicopathological data of 30 GNEN patients who were admitted to two domestic medical centers (13 cases in Wenzhou Hospital of Traditional Chinese Medicine and 17 cases in Wenzhou People′s Hospital) from January 2010 to December 2018 were collected. There were 23 males and 7 females, aged (62±4)years, with a range from 27 to 78 years. The patients underwent abdominal CT plain scan and dynamic enhancement scan. Two associate chief radiologists with more than 20 years of work experience observed and analyzed all the images respectively. Observation indicators: (1) CT examination; (2) treatment and postoperative pathological examination; (3) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect the survival of patients up to December 2018. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were described as M (range). Results:(1) CT examination: of the 30 patients, 14 had the tumor located in the fundus of stomach, 10 had the the tumor located in the body of stomach, and 6 had the tumor located in the antrum. The tumor was elliptical in 18 cases and irregular in 12 cases. There were 15 cases of endogenous type, 13 cases of exogenous type, and 2 cases of intramural type. Patients with G1 neuroendocrine tumor had the maximum diameter of (6.8±1.6)cm, of which 4 cases had the maximum diameter less than 5.0 cm and 4 cases had the maximum diameter of 5.0 to 10.0 cm. Patients with G2 neuroendocrine tumor had the maximum diameter of (8.3±2.7)cm, of which 1 case had the maximum diameter less than 5.0 cm, 4 cases had the maximum diameter of 5.0 to 10.0 cm, and 2 cases had the maximum diameter greater than 10.0 cm. Patients with G3 neuroendocrine carcinoma had the maximum diameter of (17.8±2.2)cm, of which 6 cases had the maximum diameter of 5.0 to 10.0 cm and 9 cases had the maximum diameter more than 10.0 cm. The tumor showed swelling growth in 14 cases and invasive growth in 16 cases. The tumor boundary was clear in 14 cases and unclear in 16 cases. CT plain scan showed homogeneous tumor density in 10 cases and heterogeneous density in 20 cases. Nine patients had iso-density in the tumor parenchymal part, and the CT value was (34.0±3.5)HU. In the 18 cases of low density, the CT value was (16.6±1.4)HU. In the 3 cases of high density, the CT value was (45.3±3.6)HU. Of the 30 patients, 21 cases had small punctate or small round necrotic cyst lesions in the tumor, 10 cases had mesenteric lymph node, peritoneum, liver metastasis and adjacent omentum invasion; 17 cases had abdominal effusion. In the CT enhancement examination, 12 cases showed mild enhancement, and the CT value was (56.5±6.3)HU; 15 cases showed moderate enhancement, and the CT value was (66.0±5.4)HU; 3 cases showed significant enhancement, and the CT value was (76.6±5.8)HU. Seven cases showed homogeneous enhancement and 23 cases had heterogeneous enhancement. There were 8 cases with tortuous vessels. (2) Treatment and postoperative pathological examination: of the 30 patients, 10 cases with mesenteric lymph nodes, peritoneum, liver metastasis and adjacent omentum invasion underwent radical total gastrectomy; 14 cases without surrounding tissue invasion or metastasis underwent radical subtotal gastrectomy; 6 cases with tumor diameter less than 4.0 cm and without surrounding tissue invasion or metastasis underwent endoscopic resection. All the 30 patients were confirmed GNEN by postoperative pathological examination, including 8 cases of G1 neuroendocrine tumor, 7 cases of G2 neuroendocrine tumor, and 15 cases of G3 neuroendocrine carcinoma. Results of immunohistochemical staining showed that 30 patients were positive for synaptophysin, 23 were positive for chromogranin A, and 9 were positive for cytokeratin. (3) Follow-up: all the 30 patients were followed up for 10-80 months, with a median follow-up time of 39 months. The 5-year survival rate of 30 patients was 43.3% (13/30). The 5-year survival rates were 6/8, 3/7 and 4/15 of patients with G1 neuroendocrine tumor, G2 neuroendocrine carcinoma, and G3 neuroendocrine carcinoma.Conclusions:GNEN has the main manifestation as abdominal pain, with G3 as pathological classification, which is common in fundus and body of stomach. The CT findings of GNEN are characterized by swelling or infiltrating growth and round or irregular low-density masses. Tumors are prone to cystic transformation, and showed the mildly to moderately heterogeneous enhancement.

This study was aimed to explore the effects of expressing eukaryotic elongation factor 1A1 (eEF1A1) on proliferation and apoptosis in human acute T lymphocytic leukemia (T-ALL) cell line Jurkat with knocked down eEF1A1 gene and its mechanisms. eEF1A1-expressing lentivirus (LV) was constructed and used to transfect the Jurkat cells with knocked down eEF1A1 gene. Then, the expressions of eEF1A1 mRNA and protein were detected by real time PCR(RT-PCR) and Western blot respectively.Cell proliferation, apoptosis and cycle were detected by MTT method, Annexin V-APC labeling and DNA ploidy analysis respectively. The related protein expressions of phosphatidylinositol-3-kinase (PI3K)/serine/threonine kinase (Akt) signaling pathway were detected by Western blot. The results indicated that eEF1A1 mRNA and protein expressions of Jurkat cells with knocked down eEF1A1 gene were re-established by constructing eEF1A1-expression LV. Compared with negative control group (transfected with negative control LV and eEF1A1-shRNA LV), cell proliferation in eEF1A1 expression group was significantly enhanced, cell apoptosis was remarkably inhibited, percentage of cells in G0/G1 phase was significantly reduced alone with increased percentage of cells in S and G2/M phase, and the expression levels of p-Akt (Ser 473), nuclear factor kappa B (NF-κB), p-NF-κB (Ser 468), mammalian target of rapamycin (mTOR) and p-mTOR (Ser 2448) protein significantly increased. It is concluded that eEF1A1 may have a carcinogenic effect in T-ALL cells. eEF1A1 expression has noticeable effects on the proliferation enhancement and apoptosis inhibition of Jurkat cells, which may be mediated by the up-regulation of PI3K/Akt/NF-κB and PI3K/Akt/ mTOR signaling pathway.
Apoptosis ; Cell Proliferation ; Gene Expression ; Humans ; Jurkat Cells ; Peptide Elongation Factor 1 ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Small Interfering ; genetics ; Signal Transduction

Based on the sequence of BAC (Bacterial Artificial Chromosome) along with the Cre/lox P system, the gene-targeting vectors to multiple loci of the repetitive internal transcribed spacers between rDNA genes in Leghorn chicken were constructed. The key material of multiple loci gene targeting in vivo would be obtained. First, the plasmid of pYLSV-TDN with TK, HRDS2, and Neo genes was constructed. The TK-HRDS2-Neo DNA fragment obtained from the plasmid of pYLSV-TDN was digested by Not I/HindIII and inserted into the upstream of the lox P site of BAC plasmid for obtaining the selective vector of BAC-TDN. The expression vector of pYLVS-GID with EGFP, hIFN genes, and HRDS1 was then obtained. The plasmid of BAC-TDN-VS-GID was obtained by cotransformation of the selective vector of BAC-TDN and the expression vector of pYLVS-GID to E. coli NS3529 through the action of Cre/lox P system. The gene-targeting vector of BAC-TDN-GID to multiple loci of the ITS region in Leghorn chicken was obtained by cleaving the sequence of pYLVS with the homing endonuclease of I -Sce I and ligating with the linker of LS. The insertion and the insert direction of DNA fragments were identified by restriction digestion or PCR and sequencing in each clone. The significance of the technique ofgene-targeting vector to multiple loci are shown as follows. First, the targeting loci were increased to 100 - 300. Second, the problems of unstable expression of inserted genes were partially solved. Third, the need for safety against toxicity integration was resolved. Fourth, the forbidden zone of gene integrating on the repetitive DNA sequences was broken through.
Animals ; Attachment Sites, Microbiological ; genetics ; Chromosomes, Artificial, Bacterial ; genetics ; Cloning, Molecular ; DNA ; genetics ; metabolism ; DNA Restriction Enzymes ; metabolism ; DNA, Ribosomal Spacer ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Integrases ; genetics ; Interferon-gamma ; genetics ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; Transformation, Genetic

OBJECTIVETO establish a RP-HPLC method for the determination of calycosin-7-O-beta-D-glucopyranoside in Radix Astragali, and to analyse the calycosin-7-O-beta-D-glucopyranoside content of ten samples of Radix Astragali, collected from different regions.

METHODA Polaris C18(250 mm x 4.6 mm, 5 microns) column was used and a mixture of methanol-water (30:70) was used as the mobile phase at a flow rate of 1.0 mL.min-1. The column temperature was 25 degrees C and the UV detection wavelength was 254 nm.

RESULTThe calibration curve was in good linearity over the range of 0.0106-2.12 micrograms with the regression equation Y = 3035. 97 X - 14.85(r = 0.9999). The average recovery was 95.8% (n = 5, RSD = 1.3%).

CONCLUSIONThe method is simple, quick, sensitive and reproducible. In all of the samples, the calycosin-7-O-beta-D-glucopyranoside contents differ markedly.

Astragalus membranaceus ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Ecosystem ; Glucosides ; analysis ; Isoflavones ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Species Specificity

OBJECTIVETo study hPARP1 genetic polymorphism in southern Chinese Han and Miao populations.

METHODSBlood samples from 187 and 210 southern healthy Han and Miao populations were collected. The mutations of exons 12,13,16 and 17 of hPARP1 gene were investigated by PCR-single-strand conformation polymorphism(SSCP).

RESULTSFragments of 253 bp,313 bp,175 bp,362 bp within exons 12,13,16 and 17 respectively of hPARP1 gene were amplified by multiple PCR. An SSCP variant in exons 12,13,16 and 17 of PARP1 gene in 187 healthy Han and 210 healthy Miao individuals was identified. Seven single-base substitutions compared with the sequence of PARP1 gene were identified: a T to C transition in exon 12 (Phe548Ser), a G to T transition in exon 13 (Ala683Ser), a G to T transition in exon 16 (Asp798Tyr), and a A to G transition in exon 17 (His808Arg).

CONCLUSIONThere were polymorphism sites in exons 12,13,16,17 of hPARP1 gene in southern Chinese Han and Miao populations; these results may be useful for the establishment of PARP1 genotyping, and these newly described PARP1 alleles would be advantageous indicators for population studies.

Adult ; Alleles ; China ; Exons ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; genetics ; Polymorphism, Single-Stranded Conformational

OBJECTIVESTo investigate prognostic effect of postoperative resection-margin status for intraoperatively positive resection margin in advanced gastric cancer and discuss the treatment choice for intraoperatively positive resection margins.

METHODSA retrospective study was investigated in 64 advanced gastric cancer patients with positive resection margin after potentially curative resection. The survival between 50 patients who was re-excised to a negative resection margin (NR group) and 14 patients who were left with positive resection margin (PR group) was compared. Prognostic factors were analyzed using univariate and multivariate Cox regression model analysis.

RESULTSThe median survival in the PR group was 17.0 months (95%CI: 11.6 - 22.4) as compared with 23.0 months (95%CI: 20.5 - 25.5) in the NR group (P = 0.045). However, resection-margin status lost significance on multivariate analysis. In the subgroup of D2 lymphadenectomy, the median survival in the PR group and NR group were 17.0 months (95%CI: 12.0 - 22.0) and 24.0 months (95%CI: 19.8 - 28.1) respectively; multivariate analysis further identified resection margin status as an independent prognostic factor.

CONCLUSIONSRe-excision for intraoperatively positive margin to negative margin improves the prognosis of the patients with advanced gastric cancer, and re-excision is the first choice when intraoperative frozen section detects a positive margin. Routine frozen section of resection margin should be mandatory in all advanced gastric cancer undergoing potentially curative surgery.

Female ; Follow-Up Studies ; Frozen Sections ; Gastrectomy ; methods ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Stomach Neoplasms ; surgery

OBJECTIVETo explore the clinical outcome of unilateral pedicle screw fixation plus single cage interbody fusion through spatium intermuscular of multifidus by Quadrant system.

METHODSFrom April 2008 to April 2009, 47 patients underwent unilateral pedicle screw fixation plus single cage interbody fusion through spatium intermuscular of multifidus. There were 22 males and 25 females with the mean age of 58.2 years (range, 46-74 years). Among them 12 cases had far-lateral lumbar disc herniation, 7 cases had post-discectomy recurrence, and 28 cases had degenerative instability. Thirty-seven cases were treated with lumbar interbody fusion through transforaminal approach, 10 cases through posterior approach. After surgery, the radiography was carried out to demonstrate the fusion status, and the Nakai criterion was used for assessment.

RESULTSThe average skin incision length was 3.2 cm (range, 3.0 to 3.5 cm), the average operative time was 90 min (range, 70 to 160 min), and the average blood loss was 130 ml (range, 90 to 360 ml). All cases were followed up for 8 - 20 months (average 13.6 months). Postoperative radiography showed no evidence of instrument failure, and 43 cases got bone fusion, 4 cases got suspicious fusion. At final followed-up the average leg pain VAS decreased from 7.4 ± 1.1 preoperatively to 2.4 ± 1.3 postoperatively, the average low back pain VAS decreased from 6.7 ± 1.3 preoperatively to 1.8 ± 1.5 postoperatively. According to Nakai criterion, 31 cases were rated as excellent, 11 cases as good, and 5 cases as fair with the total excellent and good rate of 89.4%.

CONCLUSIONSUnilateral pedicle screw fixation plus single cage interbody fusion through spatium intermuscular of multifidus has some advantages of minimal invasiveness, less blood loss, less complications and reliable curative effect. It is a satisfactory lumbar fusion method under suitable indication.

Aged ; Bone Screws ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Muscle, Skeletal ; surgery ; Spinal Fusion ; methods ; Treatment Outcome

This study was purposed to explore the expression of P27(kip1)and cyclin G in patients with acute leukemia (AL) and its correlation. The reverse polymerase chain reaction (RT-PCR) was used to analyse the expression of P27(kip1) and cyclin G mRNA in 89 AL patients and 10 normal persons; Western blot was used to analyze the expression of P27(kip1) and cyclin G protein in 39 AL patients and 10 normal persons. The results showed that the cyclin G mRNA and protein expressions in new diagnosed/relapsed cases of AL were significantly higher than those in patients with remission and normal controls (p < 0.05 and p < 0.01), but there was no difference between remission cases and normal controls (p > 0.05). The expression of P27(kip1) mRNA in newly diagnosed/relapsed patients with AL was not significantly different from patients with remission and normal controls (p > 0.05), while the P27(kip1) protein expression in remission cases of AL and normal controls was significantly higher than that in new diagnosed/relapsed cases (p < 0.05), but there was no difference between remission cases and normal controls (p > 0.05). The expression of P27(kip1) negatively and lowly correlated with the expression of cyclin G in patients with AL. It is concluded that the low expression of P27(kip1) and the high expression of cyclin G in patients with AL may have some correlation with genesis and development of AL and may be an indication for poor prognosis of AL.
Adult ; Cyclin G1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male