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.

The purpose of this study was to compare the differences of the protein expression profiles between human myeloid leukemia K562 cells and adriamycin-resistant K562/A02 cells, as well as to select novel resistance-related proteins in myeloid leukemia by means of proteomics. The total cellular proteins were separated from K562 and adriamycin-resistant K562/A02 cells by using technique of two dimensional difference in gel electrophoresis (2D-DIGE). Differentially expressed proteins were analyzed by matrix-assisted laser desorption ionization/time of flight-mass spectrometry (MALDI-TOF/MS), and by protein database searching. Moreover, the differentially expressed proteins were verified at protein and mRNA levels by Western blot assay and quantitative real time PCR. The results showed that 8 proteins differentially expressed in adriamycin-resistant K562/A02 cells, among them 2 proteins were identified to be down-regulated and 6 to be up-regulated. These identified proteins involved in the cell energy metabolism, cell proliferation, cell apoptosis, signal transduction, gene transcription and translation respectively. The results assayed by Western blot were similar to those detected by 2D-PAGE. Two up-regulated proteins Stathmin and CrkL were selected for verification in K562 and K562/A02 cells. As a result, the results detected by Western blot were identical with results from 2D-DIGE; real time quantitative PCR assay showed that the changes of CrkL at mRNA level were identical with changes at protein level, but no complete identity of Stathmin changes at mRNA level and protein level was observed. It is concluded that the difference of protein expression profile exists in K562 and K562/A02 cells. Stathmin and CrkL proteins may be involved in the drug resistance and suggest a novel clue for the resistant mechanisms in myeloid leukemia, which is worth further to explore.
Adaptor Proteins, Signal Transducing ; metabolism ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Humans ; K562 Cells ; Leukemia ; metabolism ; Nuclear Proteins ; metabolism ; Stathmin ; metabolism

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

In order to clone the full-length cDNA of a novel EST which is probably related to acute leukemia relapse and to analyse the sequences, the electronic cloning technique combined with RT-PCR was used to clone the full-length cDNA, and the sequences were analyzed by bioinformatics. The results showed that the two novel splicing variants of eIF4E named as splicing variant 1 and 2 of eIF4E were obtained. Bioinformatics analysis showed that variant 1 and 2 exhibited 84% and 47% similarity to eIF4E mRNA, and were localized on eIF4E locus on chromosome 4. The lengths of 2 variants are 1 904 bp and 3 393 bp, encoding 245 amino acids and 132 amino acids respectively. BLAST results showed that the both variants mentioned above contains seven exons. Among them, the sequences of the three exons at 5' end of variant 1, variant 2 and eIF4E mRNA were different from each other. Protein BLAST showed that they are partially different from eIF4E protein. It is concluded that the two novel splicing variants of eIF4E were cloned, and their relation to acute leukemia relapse needs to be further investigated.
Base Sequence ; Cloning, Molecular ; Computational Biology ; DNA, Complementary ; genetics ; DNA, Recombinant ; Eukaryotic Initiation Factor-4E ; genetics ; Humans ; Molecular Sequence Data ; Protein Isoforms ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

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

OBJECTIVETo evaluate the effect of carvedilol on the cardiac function and autoantibodies against the cardiac beta(1), beta(2) and alpha(1)-adrenergic receptors in patients with chronic heart failure (CHF).

METHODS54 patients with CHF were divided randomly into two groups, one was regular treatment group treated with ACE inhibitor, digoxin and diuretic, another was carvedilol treatment group treated with carvedilol on the basis of above regular treatment. All the patients were followed up for six months and measured the changes of cardiac function and three autoantibodies by echocardiography and enzyme-linked immunosorbent assay (ELISA) respectively.

RESULTS(1) After carvedilol treatment, LVEDD and LVESD (57.50 +/- 7.29) mm and (43.17 +/- 8.27) mm were smaller than that in regular treatment group [(64.09 +/- 7.40) mm and (52.93 +/- 8.35) mm], and LVEF [(50.41 +/- 10.91)%] was higher than that [(41.70 +/- 7.45)%] in regular treatment group (P < 0.01). (2) After carvedilol treatment, the positive ratios and average titers of autoantibodies against the cardiac beta(1), beta(2) and alpha(1)-adrenergic receptors all decreased significantly compared with that of pre-treatment (P < 0.05). The positive ratios of autoantibodies against the three receptors in carvedilol treatment group were lower than those in regular treatment group (P < 0.05). The average titers of autoantibodies against the cardiac beta(1), beta(2) and alpha(1)-adrenergic receptors in carvedilol treatment group (1:72.44, 1:61.66 and 1:67.30) were lower than those in regular treatment group (1:113.24, 1:110.66 and 1:113.24), P < 0.05.

CONCLUSIONSCarvedilol decreased positive ratio and average titer of autoantibodies against the beta(1), beta(2) and alpha(1) receptors accompanied with the obvious improvement of cardiac function though the blockade of beta(1), beta(2) and alpha(1) receptors. It suggests that the autoantibodies might be involved in the process of pathophysiology and development of CHF. Carvedilol can inhibit this process.

Adult ; Aged ; Autoantibodies ; blood ; Carbazoles ; therapeutic use ; Female ; Follow-Up Studies ; Heart Failure ; drug therapy ; immunology ; physiopathology ; Humans ; Male ; Middle Aged ; Propanolamines ; therapeutic use ; Receptors, Adrenergic, alpha-1 ; immunology ; Receptors, Adrenergic, beta-1 ; immunology ; Receptors, Adrenergic, beta-2 ; immunology ; Young Adult

OBJECTIVETo investigate the safety and efficacy of close reduction and percutaneous needle fixation for the treatment of anterior dislocation of sternoclavicular joint.

METHODSA retrospective analysis was performed with 6 cases of anterior sternoclavicular dislocation by close reduction and percutaneous needle fixation with Kirschner wire treated from January 2001 to February 2009, including 5 males and 1 female aged from 19 to 45 with an average of 28.8 years old. Among the 6 cases, 4 were on right lateral and 2 were on left lateral. The time from injured to treatment was from 6 hours to 12 days (averaged 4.5 days). The clinical effects were evaluated according to Rockwood scoring and the complications were observed.

RESULTSAll 6 patients were followed-up for 3 to 13 months (averaged 6 months). According to Rockwood scoring,the preoperative score was (7.00 +/- 0.89) points, postoperative score was (13.17 +/- 1.72) points; the results showed excellent in 5 cases and good in 1 case. No local infection, postoperative pain,recurrent dislocation,broken needle, and other complications were observed in this study.

CONCLUSIONThe treatment of anterior sternoclavicular joint dislocation with Kirschner wire minimally invasive fixation is an easy, reliable fixation with less complications.

Adult ; Bone Wires ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Joint Dislocations ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Radiography ; Sternoclavicular Joint ; injuries ; 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