Purpose This subject detected the expression of paper miRNA34a,c-myc gene and protein in adenocarcinoma of lung tissue,adjacent mucosa tissue and normal lung tissue exploring the relationship between the two and the clinical significance.Method The project mainly used the method of real-time fluorescent quantitative PCR (qRT-PCR),Western blot and immunohistochecical to detect the expression of miRNA34a and c-myc in normal tissue and adenocarcinoma of lung respectively.Result (1) lower expression of miRNA34a in lung adenocarcinoma tissue,lower than the adjacent and normal tissues (P ＜ 0.01),higher than the metastatic carcinoma group (P ＜ 0.01).C-myc is highly expressed in lung adenocarcinoma tissue,higher than the adjacent and normal tissues (P ＜0.01),lower than the metastatic carcinoma tissue (P ＜ 0.01).miRNA34a and c-myc beside carcinoma tissues and normal tissues also have significant difference between the statistical significance (P ＜ 0.05).(2) Differentiation of low and high there was no statistically significant difference between patients with lung adenocarcinoma (P ＞ 0.05).there were significant differences in the expression of miRNA34a and c-myc in the tissues of metastatic and non metastatic adnocarcinoma.(3)The positive expression of c-myc group of lung cancer patients was lower than that of the negative group (P ＜ 0.05),low expression of the survival time in miRNA34a group was significantly lower than that in high expression group,there was significant difference (P ＜ 0.01).Conclusion Expression of miRNA34a and c-myc both showed a negative correlation in adenocarcinoma of lung.

[ ABSTRACT] AIM: To detect basic fibroblast growth factor ( bFGF ) expression in clinical common malignant tumor ( non-small-cell lung cancer,breast cancer, colon cancer and melanoma) , and to identify relationship between the expression and tumor clinicopathological characteristics.METHODS:Immunohistochemical SP method was used to detect the expression of bFGF at protein level in 208 cases of paraffin-embedded tissue of primary malignant tumor patients ( 68 cases of lung cancer, 80 cases of breast carcinoma, 41 cases of colon cancer and 19 cases of melanoma) .RESULTS:The bFGF protein expression levels were significantly higher in low differentiated non-small-cell lung cancer with lymph node metastasis, and were positively correlated with TNM.In addition, no significant influence of the bFGF protein expression on the patients with median survival period was observed.The protein expression of bFGF was higher in advanced breast cancer with lymph node metastasis and was commonly found in the middle/higher differentiated colon cancer with regional lymph node metastasis.Meanwhile, bFGF protein was highly expressed in advanced melanoma patients with lymph node metastasis.CONCLUSION:bFGF may participate in the process of occurrence and progression of malignant tumor.Ex-pression of bFGF protein may be an effective parameter for evaluating metastasis and prognosis of malignant tumor.

AIM:To detect the expression of basic fibroblast growth factor (bFGF) and matrix metalloprotei-nase 9 (MMP9) in nasopharyngeal carcinoma (NPC) and its correlation with clinicopathological features and prognosis of the patients.METHODS:The expression of bFGF and MMP9 was detected by the method of SP immunohistochemical staining in biopsy tissues of NPC patients.The relationship between the expression and the clinical significance was analyzed as well.RESULTS:In 289 cases of NPC patients, the positive rates of bFGF and MMP9 were 71.3% and 61.6%, respectively.Correlation analysis demonstrated that the expression rates of bFGF and MMP9 were both positively associated with N stage and clinical stage in NPC patients.The high expression rates of both bFGF and MMP9 were associated with poor overall survival and progression-free survival of NPC patients.Furthermore, the positive rate of bFGF was positively correlated with that of MMP9, and over-expression of both bFGF and MMP9 was correlated with the poorest survival outcomes in NPC patients.CONCLUSION:bFGF and MMP9 are over-expressed in NPC tissues and significantly associated with NPC recurrence and poor outcome.The combined interpretation of bFGF and MMP9 expression levels leads to refinement of the risks for the NPC patients and could be chosen as the prognostic biomarkers.

Objective To observe the impacts of Xingnao Kaiqiao acupuncture(AC)method on synaptic plasticity and intestinal flora in stroke rats.Methods The middle cerebral artery occlusion method was applied to establish a rat model of ischemic stroke,and the rats with Zea Longa score of 1-3 were randomly grouped into model(M)group and AC group,and rats without middle cerebral artery occlusion were regarded as the sham(S)group,with 15 animals per group.After modeling,groups S and M were not intervened,and group AC was intervened with AC,once a day,for 7 consecutive days.The Zea Longa method was applied to assess neurological function;transmission electron microscopy(TEM)and Golgi staining were applied to visualize synaptic structures in the ischemic penumbra(IP);Western blot was applied to detect the protein expression of BDNF,SYN and GAP-43 in the IP cortex tissue;ELISA was applied to detect the content of IL-1β,TNF-α,IL-17 in the IP cortex tissue;16S rDNA amplification and sequencing method was applied to analyze the structure of rat intestinal flora.Results Compared with the S group,the M group had irregular synaptic morphology,blurred boundaries,thinned postsynaptic zona densities,higher neurological function scores,protein levels of BDNF,SYN,GAP-43 and content of IL-1β,TNF-α,IL-17(P<0.05),and lower density of spines(P<0.05).Compared with the M group,AC intervention could improve the synaptic morphology,reduce the neurological function score and content of IL-1β,TNF-α,IL-17(P<0.05),and increase the density of dendritic spines,BDNF,SYN,and GAP-43 protein levels(P<0.05).The intestinal flora analysis showed that compared with the S group,the M group had a reduced diversity and number of bacterial flora,at the phylum level,the M group had a lower relative abundance of Bacteroidetes,and higher F/B ratio(P<0.05);at the genus level,the group M had a higher relative abundances of Bacteroides and Rikenbacteria(P<0.05),and lower relative abundances of Lactobacillus and Lachnospiraceae_UCG-006(P<0.05).Compared with the M group,AC intervention could increase the diversity and quantity of the flora,increase the relative abundance of Bacteroidetes and reduce the F/B ratio at the phylum level(P<0.05),increase the relative abundances of Faecalibacterium and Lactobacillus and reduce the relative abundances of Riken,Escherichia-Shigella,etc.at the genus level(P<0.05).Conclusion Xingnao Kaiqiao acupuncture can restore the relative abundance of beneficial bacteria,reduce the relative abundance of harmful bacteria,attenuate brain inflammation,improve synaptic plasticity,and reduce neurological damage in stroke rats.

AIM　To investigate the effect of frozen recombinant staphylokinase on the hemostatic and fibrinolytic systems in healthy volunteers, in order to obtain reliable evidence for the possibility of further clinical application. METHOD　r-Sak had been taken intravenously by 20 cases of healthy volunteers in different dosages (1 mg, 2.5 mg, 5 mg, 10 mg, 15 mg). The clinical hemorrhagic manifestations were observed and a set of hemostatic tests(BT, BPC, ATPP, PT, TT, Fg) and fibrinolytic tests (PL∶A,α2-PI∶A, FDP, D-D) monitored before and after injection.　RESULT　Four of 20 volunteers showed slight hemorrhagic tendency on mucocutaneous area (3/4 from gingivea and 2/4 at the sites of injection). It stopped spontaneously. None of them showed visceral bleeding. There were no significant changes in hemorrhagic and coagulative phases. Only 4 of them showed slight abnormal changes in D-D. It was supported that r-Sak was a highly selective fibrirolytic agent without significant influence in human hemostatic and coagulatic system. CONCLUSION　The specific ranges of doseges, r-Sak is a relatively safe and well tolerated agent for healthy people. Further clinical study is still needed for the suitable dosage for clinical application.

OBJECTIVESTo summarize the experience in the treatment of 112 cases of complex bone nonunion from 1982 to 1999 in our department and introduce the technique of external skeletal fixation.

METHODSThe two fragment ends of all cases were fixed under pressure with half-ring sulcated external skeletal fixator. Those cases complicated by bone defect or limb shortening were operated on with epiphysiotomy to restore the length of the limb in the period of compressive fixation or after the occurrence of bone union according to the condition of complicated infection and the length of the limb shortened.

RESULTSThe nonunion of the 112 cases was united eventually. The infection in 34 cases was eradicated. Bone union in cases without infection took 3 approximately 7 months (average 5.2 months) and in cases with infection took 5 approximately 11 months (average 5.5 months). The length of the limb in 11 cases with bone defect was restored in the same period of compressive external fixation and another 8 cases achieved after bone union. The length between the injured and healthy limbs was balanced.

CONCLUSIONSWhen external skeletal fixation is employed to treat those troublesome cases of bone nonunion, the pins for fixation are inserted in sites far from the lesions and the non-united fragment ends are exposed only in the area without scars. Consequently, there is little interference with the blood circulation and the osteogenic potency of the fragment ends. The sclerotic bone tissue is not excised, the marrow cavity is not chased to be open and the fragment ends are only moderately modified. As a result, the stability of fixation is increased and further shortening of the limb avoided. External skeletal fixation using small pins with cross penetration results in plastic fixation and promotes bone healing. Bone lengthening with epiphysiotomy can restore the balance of the limbs.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Fracture Fixation ; Fracture Healing ; Fractures, Ununited ; Humans ; Male ; Middle Aged

Background:Cerebellar fastigial nucleus (FN)is involved in regulation of visceral activities such as cardiovascular, ingestion,respiratory,and acute gastric mucosal injury,yet it is unclear whether it participates in the regulation of visceral hypersensitivity and what is the possible mechanism. Aims:To investigate the effect and possible mechanism of glutamic acid (Glu ) injection into cerebellar FN on chronic visceral hypersensitivity in rats. Methods: Chronic visceral hypersensitivity rat model was established by neonatal colorectal distension (CRD). After 8 weeks,the rats were divided into CRD group,solvent group (0. 2 μL 0. 9% NaCl solution injection into cerebellar FN),high-,medium-,low-dose Glu groups (12,6,3 μg Glu injection into cerebellar FN,respectively),3-MPA +Glu group (12 μg Glu injection after glutamate decarboxylase inhibitor 3-MPA injection into cerebellar FN),Bic + Glu group (12 μg Glu injection into cerebellar FN after GABAAreceptor blocker Bic injection into lateral hypothalamic area). Pain threshold,abdominal withdrawal reflex (AWR)score and abdominal external oblique muscle electromyography (EMG)were used to detect visceral sensitivity,and malondialdehyde (MDA)content and superoxide dismutase (SOD)activity were measured. Results:Chronic visceral hypersensitivity rat model was successfully established. Compared with CRD group,pain threshold was significantly increased (P＜0. 05),AWR score,EMG amplitude,MDA content were significantly decreased (P＜0. 05 ),and SOD activity was significantly increased in a dose-dependent manner in Glu group (P ＜0. 05 ). Compared with 12 μg Glu group,pain threshold was significantly decreased (P＜0. 05),AWR score,EMG amplitude, MDA content were significantly increased (P ＜0. 05),and SOD activity was significantly decreased in 3-MPA +Glu group,Bic+Glu group (P＜0. 05). Conclusions:Glu injection into cerebellar FN can significantly reduce the visceral sensitivity in rats. The mechanism may be that Glu in cerebellar FN produces GABA via glutamate decarboxylase,and then binding GABAAreceptor in lateral hypothalamic area,resulting in increased intestinal mucosal antioxidant capacity, thereby reducing visceral hypersensitivity.

Objective: To investigate the effects of glutamate (Glu) injected into hypothalamic paraventricular nucleus (PVN) on visceral pain of chronic visceral hypersensitivity (CVH) rats and its possible mechanism. Methods: Newborn SD rats were given CVH rat model by colorectal distension (CRD) on the 8th, 10th and 12th day after birth. Thirty rats with successful CVH model were randomly divided into CVH model group (CVH group), CVH + injection of saline into PVN group (NS group), CVH+ injection of Glu into PVN (3, 6, and 12 μg Glu, namely G3, G6, and G12, respectively), 6 rats in each group, and 6 SD rats with matching body mass were taken as sham operation group (Sham group). The pain behavior of the rats was evaluated by pain threshold, abdominal withdrawal reflex (AWR) score, and abdominal external oblique muscle electromyography (EMG). The expression of arginine vasopressin (AVP) and the proliferation of colon tissue were detected by immunohistochemical staining. The apoptosis of colon tissue was detected by TUNEL. Results: Compared with the NS group, the pain thresholds of the G3, G6 and G12 groups increased, and the AWR scores and EMG amplitudes decreased. The differences were statistically significant(Pain threshold: t=7.65, 16.31, 24.78, both P<0.05; AWR scores: t=-2.98, -4.77, -7.29, both P<0.05; EMG amplitudes: t=-3.06, -5.75, -8.92, both P<0.05). Compared with the Sham group, the expression of AVP in PVN of the CVH group and NS group decreased ((42.63±5.20) %, (18.67±2.94) %, (17.53±2.47) %; t=6.95, t=7.56, both P<0.05). The expression of AVP was increased after different doses of Glu into PVN, and the AVP level in G12 group ((18.15±6.49)%) was higher than that of NS group, the difference was statistically significant (t=-4.21, P<0.05). Compared with the Sham group, the expression of PCNA in colonic mucosal cells of the CVH group and NS group decreased ((65.48±1.68) %, (18.39±1.67) %, (17.69±1.68) %; t=34.35, t=34.80, both P<0.05). The expression of PCNA was increased after different doses of Glu injected into PVN, and the PCNA level in G12 group ((59.91±5.63)%) was higher than that of NS group, the difference was statistically significant (t=-12.44, P<0.05). Compared with the Sham group, the expression of apoptotic cells in colonic mucosal cells of the CVH group and NS group increased ((23.38±11.40)%, (83.79±3.57)%, (80.91±2.47)%; t=-8.77, t=-8.54, both P<0.05). The expression of apoptotic cells was decreased after different doses of Glu into PVN, and the G12 group was ((18.15±6.49) %). Compared with NS group, the difference was statistically significant (t=15.65, P<0.05). Conclusion Injection of Glu into hypothalamic PVN can alleviate the visceral pain behaviors in CVH rats, and its mechanism may be related to arginine vasopressin.

OBJECTIVETo investigate the characteristics of the abnormalities of chromosome 17 in myeloid malignancies with complex chromosomal abnormalities (CCAs).

METHODSAbnormalities of chromosome 17 were analyzed in 73 patients with myeloid malignancies with CCAs showed by R banding and conventional karyotyping, including 21 acute myeloid leukemia (AML), 36 chronic myeloid leukemia (CML) and 16 myelodysplastic syndrome (MDS). All CCAs were further analyzed by multiplex fluorescence in situ hybridization (M-FISH).

RESULTSAmong the 73 myeloid malignancies with CCAs, chromosome 17 was the most frequently involved chromosome. It was found in 46.5% (34/73) of all cases, including 12 AML, 13 CML in blast crisis (BC) and 9 MDS. However, it was not found in the 9 CML cases in chronic phase (CP). The majority of changes were structural rearrangements which were identified in 43.8%(32/73)of all cases, among them the frequency was 52.4% (11/21), 33.3% (12/36) and 56.3% (9/16) in AML, CML and MDS, respectively. Numerical abnormalities were detected in 15.1% (11/73) cases, all were monosomy 17, and the frequency was 25.0% (3/12), 38.5% (5/13) and 33.3% (3/9) in AML, CML and MDS, respectively. Both numerical and structural abnormalities of chromosome 17 were found in 9 cases. Unbalanced translocations involving chromosome 17 were much more frequent than balanced ones. In the 3 groups, 16, 15 and 8 unbalanced translocations were found respectively. Only two kind of balanced translocations including t(15;17) in AML and t(15;17;22) in CML were found. All chromosomes were involved except chromosomes 5, 6 and 22 as partner chromosomes, the most common one was chromosome 15 (8.2%), followed by chromosome 2 (5.4%). Five of the 6 cases with translocation of chromosomes 15 and 17 were acute promyelocytic leukemia, the other case was CML-BC.

CONCLUSIONAbnormalities of chromosome 17 were the most frequently involved chromosomal aberrations in myeloid malignancies, and structural rearrangements were more common. All the numerical abnormalities were monosomy 17, unbalanced translocations were much more frequent than balanced ones.

Adolescent ; Adult ; Aged ; Child ; Chromosome Aberrations ; Chromosomes, Human, Pair 17 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics

Objective To evaluate the analgesic effect of tetrodotoxin (TTX) in four types of acute pain models and provide experimental support for its rational application. Methods Mice or rats were intramuscularly pretreated with morphine (1 mg/kg) or TTX (0, 0.5, 1, 2, 4 and 8 μg/kg) 40 min before acetic acid writhing test, formalin stimulation test, hot plate test or tail flick test. Pain response or pain threshold were recorded, and inhibition rate was calculated during the tests. The arachidonic acid of serum was determined by Elisa. Results Significant analgesic effects were observed with morphine in all four acute pain models. TTX dose-dependently reduced the number of writhing induced by acetic acid and inhibited the pain response induced by formalin during phase I and phase II, with the highest inhibition rate of more than 80.00% in two pain models. TTX showed analgesic effect in tail flick test and hot plate test, with the highest inhibition rate of 25.00% and 19.79%, respectively. Both acetic acid and formalin increased arachidonic acid in animal serum, but TTX had no significant inhibitory effect on the releasing of arachidonic acid. Conclusion TTX showed significant analgesic effect in the chemical stimulation pain models induced by acetic acid and formalin, but limited analgesic effect was observed on the physical stimulation pain model induced by heat (hot plate and hot water). TTX may produce analgesic effect by blocking the inflammatory mediators mediating pain response.