BACKGROUND: Chronic obstructive pulmonary disease(COPD) is characterized of chronic inflammation in airway, pulmonary parenchyma and pulmonary vessels. The mechanism of the increment and activity changes of these inflammatory cells is unclear at present.OBJECTIVE: To study the apoptotic character of peripheral blood polymorphonuclear neutrophils(PMNs) and its relationship with COPD to provide a reference for early intervention and function surveillance for COPD patients.DESIGN: An observatory comparative study based on COPD patients and healthy population as controls.SETTING: Department of pulmonary medicine in a military medical university of Chinese PLA affiliated hospital.PARTICIPANTS: Totally 98 COPD patients were admitted by the Department of Pulmonary Medicine, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA between February 2003 and December 2003 due to COPD acute attack. Eighteen patients including 12 males and 6 females aged between 48 and 70 years old [mean of(56 ± 7) years old]were randomly selected into COPD group according to random number table.Totally 14 healthy adults including 10 males and 4 females aged between 50 and 70 years old [mean of (59 ± 8) years old] who were individuals came for physical check up in our hospital were selected in control group.METHODS: PMNs were separated from peripheral blood by density gradient centrifugation. The dynamic changes of PMNs apoptosis in peripheral blood was observed by flow cytometer and TUNEL method.MAIN OUTCOME MEASURES: Comparison of PMNs apoptotic rate in peripheral blood among groupsRESULTS: As indicated by flow cytometric analysis, PMNs apoptotic rate at early apoptotic phase in COPD patients at paracmasis was(8.5 ± 1.3)%,which was significantly lower than(12.5 ± 1.8)% of normal control group( t=6.25, P ＜ 0. 01); PMNs apoptotic rate was(5.1 ±0.6)% at acute aggravation stage, which was significantly lower than that of paracmasis group ( t =5.66, P ＜001). As indicated by TUNEL analysis, PMNs apoptotic rate at paracmasis was(12.42 ±2.7)% , which was significantly lower than (21.5±4.8)% of normal control group(t=5.76, P ＜ 0.01); PMNs apoptotic rate was(4. 9 ±0.4)% at acute aggravation stage, which was significantly lower than that of paracmasis group( t = 6. 12, P ＜ 0. 01 ) . PMNs changes at the late phase of apoptosis/necrosis had a contrary tendency, i. e.,PMNs rate at late apoptotic phase/necrosis was(2. 8 ± 0.5)% in COPD patients at paracmasis, which was significantly higher than(1. 3 ±0.4)% of normal control group ( t= 6. 37, P ＜ 0. 01 ); PMNs rate was (3.7 ± 0. 3) % at acute aggravation stage, which was significantly higher than that of paracmasis group(t=5.81, P ＜0.01).CONCLUSION: PMNs abnormal apoptosis might be an important reason that induces PMNs aggregation in airway and lung tissues in COPD. This process might have an important significance in the generation and development of chronic airway inflammation, which provides an etiologic basis for the primary rehabilitative intervention of COPD.

Objective To evaluate the application of CT perfusion in the assessment of muscle perfusion in ischemic rabbit hindlimb treated by implantation of bone marrow mesenchymal stem cells.Methods Twenty rabbits were divided into two groups randomly (each n =10).Bone marrow mesenchymal stem cells were injected into the left ischemic hind limbs of animal models of intravenous injection group,with same volume of normal saline injected into the control group.Twenty-eight days later,the collateral circulation and blood vessel neogenesis were examined by Toshiba Aquilion One 320-MSCT and pathological approaches.t test and Pearson test were used to compare the parameters.Results The results show that rAF,rBV,rClearance,rMVD were 1.15 ±0.67,1.19 ±0.32,0.62 ±0.20,and 1.34 ±0.28 in intravenous injection group and 0.57 ±0.17,0.74 ±0.19,2.06 ±0.15,0.62 ±0.19 in the control group respectively.There was significant difference of rAF,rBV,rClearance and rMVD between the intravenous injection group and the control group(t =5.75,5.01,-2.81,6.43 respectively,P ＜ 0.01).There was significant correlation between rBV,rAF,rC and rMVD (r =0.857,0.811,0.615,P ＜ 0.01).Conclusion CT perfusion imaging is a relatively accurate technique to assess changes of muscle perfusion in ischemic rabbit hindlimb treated by implantation of bone marrow mesenchymal stem cells.

Objective To study the risk factors of intracranial hemorrhage after endovascular therapy of cere-bral arteriovenous malformations embolization.Methods the clinical records and images of 230 patients with cAVMs were retrospectively reviewed.Patients with intracranial hemorrhage after endovascular therapy in 3 days were recorded and the risk factors of hemorrthage by using multivariate logistic regression for guiding the treatment strategy were studied.Results There were 15 patients occurred intracerebral hemorrhages.After treatment the area of cAVMs was significantly smaller than that before treatment[(2.18 ±0.91)cm vs (6.67 ±1.56)cm],the difference lvas a statis-tically significant (t =9.627,P <0.05).A partial AVM reduction of ≥36.5%(P =0.031)and hypertension(P =0.025)were considered as risk factors for hemorrhage after endovascular therapy of cAVWs.Conclusion Patients receives a partial AVMreduction of ≥36.5% or with hypertension history have a tendancy of increasing hemorrhage after endovascular therapy of cAVMs.It's helpful for preventing intracerebral hemorrhage if the arterial blood pressure maintains after endovascular therapy.

Objective To evaluate the effects of fluid resuscitation and large-volume hemofiltration (HVHF) on the Alveolar-arterial oxygen exchange in patients with refractory septic shock. Method A total of 89 intensive care patients with refractory septic shock treated with fluid resuscitation and/or HVHF were enrolled between August 2006 and December 2009. All the patients were randomly divided into two groups. In group A, patients were treated with fluid resuscitation, n = 41 cases) and in group B, patients were treated with large-volume hemofiltration and fluid resuscitation, n =48). The O2 content of central venous blood(CcvO2), arterial oxygen content (CaO2), Alveolar-arterial oxygen pressure difference (P(A-a)DO2), the ratio of arterial oxygen pressure/alveolar oxygen pressure (PaO2/PAO2), respiratory index (RI) and oxygenation index (OI) were checked. The levels of oxygen exchange in two groups were detected by arterial blood gas analysis before treatnent, 24 hour, 72 hour and 7 days after treatment. The APACHE Ⅱ scores in patients with refractory septic shock were measured before and the 7th day after treatment with HVHF and/or fluid resuscitation respectively. Data were analyzed by using t -test and chi-square test to compare the differences and ratio between two groups and were expressed in mean ± standard deviation, and the analysis of variance was done with SPSS version 12.0 software. Results ① The differences in CcvO2 and CaO2 between two groups were[(0.60±0.24) vs. (0.72±-0.28), P ＜0.05 and (0.84±0.43) vs. (0.94±0.46), P ＜0.05]; and the oxygen extraction rates (O2ER) were significantly different between two groups [(28.7±2.4) vs. (21.7±3.4), P<0.01];② The levels of P(A-a)DO2、ratio of PaO2/PAO2、RI and OI in group B were reduced more significantly than in group A (P<0.05 or P<0.01);③The APACHE Ⅱ scores in both groups were gradually reduced after treatment for 7 days, and the APACHE Ⅱscore in group B on the 7th day of treatment were lower than that in group A[(17.2 ± 6.8) vs. (8.2 ± 3.8), P ＜ 0.01]. Conclusions Fluid resuscitation and HVHF could improve alveolar-arterial-oxygen exchange in patients with refractory septic shock, and at the same time decreased the APACHE Ⅱ scores, improving the survival rate of patients.

Aim To study the function of VR1 in chronic pain, to construct VR1 siRNA expression vectors and to study their silencing effect in the DRG neurons of rats were detected.Methods The hairpin sequences of siRNAs targeting VR1 gene of rat were designed, and two pairs of oligonucleotide sequence were synthesized. The annealed oligonucleotide fragments were cloned into linearized pRNAT-U6.2/Lenti expression vector and identified by PCR and DNA sequencing.Then, they were co-transfected by lipofectamine into 293T cells.The silencing effects of the lentivector-mediated VR1 siRNAs on the expression of VR1 mRNA were determined by RT-PCR after intrathecal injection in rats.Results DNA sequencing showed that the oligonucleotide fragments were correctly cloned into linearized pRNAT-U6.2/Lenti expression vector and the expression of VR1 mRNA in L4-L6 DRG neurons was inhibited significantly by pRNAT-U6.2/Lenti-siVR1 after intrathecal injection in rats.Conclusion The lentivector-mediated siRNAs are successfully constructed and they inhibit the expression of VR1 mRNA in the DRG neurons of rats, which may provide a potential tool for the further study and treatment of chronic pain.

AIM:To investigate the mechanism of 9-cis-RA inhibiting the growth of lung adenocarcinoma cells, and we detect the expressional changes of cyclinD1 and cdk4 in lung adenocarcinoma cells PG, A_(549), SPC-A_1 before and after being treated with 9-cis-retinoic acid(9-cis-RA). METHODS: RT-PCR was used to analyse the transcriptional changes of cyclinD1 and cdk4 in PG, A_(549) and SPC-A_1. RESULTS:9-cis-RA decreased the transcription of cyclinD1 in PG and A_(549)(P

Objective To investigate clinical value of endoscopic ultrasonography (EUS) on diagnosis and treatment of esophageal leiomyoma.Methods The clinical feature of patients diagnosed as esophageal leiomyoma by EUS was analyzed.All patients underwent conventional gastroscopy and EUS examination as well as blood test.The EUS findings were compared with histopatholoical diagnosis for endoscopically or surgically resected specimens.All patients were followed-up with EUS for 14.3 months (range 2-36 months) for leiomyoma recurrence.Results A total of 191 patients were diagnosed with esophageal leiomyomas by EUS.Clinical manifestation,serum examination,conventional gastroscopy and spiral CT were not helpful for diagnosing esophageal leiomyoma,One hundred and sixteen patients with esophageal leiomyomas were treated by either endoscopic resection (111 cases) or surgical excision (5 cases).of which,101 resected specimens were pathologically confirmed as leiomyomas.The diagnostic accuracy of EUS was 87%.The follow-up study showed that there was no change of leiomyomas in 75 untreated patients and no recurrence was found in 116 surgically treated patients.Conclusion Esophageal leiomyoma is a benign tumor,EUS plays a very important role in diagnosis and treatment of this disease.

This study is to observe the effect of N-(3-phenylallylidene)-6-fluoro-1, 8-(2, 1-propoxy)-7-(4-methylpiperazin-1-yl)-quinolin-4(1H)-one-3-carbonyl hyarazine (FQ16) on apoptosis of hepatocarcinoma SMMC-7721 cells in vitro. With different concentrations of FQ16 at different times used to treat SMMC-7721 cells in vitro, the proliferation of the cells and the inhibition effect of FQ16 on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by Hoechst 33258/PI fluorescence staining, TUNEL and agarose gel electrophoresis method. The effect of FQ16 on topoisomerase II activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Mitochondrial membrane potential (MMP, delta psi m) was measured by high content screening image system. The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression changes of Bcl-2 mRNA and Bax mRNA. The caspase-9, caspase-8, caspase-3, p53, Bcl-2 and Bax protein expressions were detected by Western blotting analysis. The results showed that the cell proliferation was inhibited by FQ16 at 0.625 - 10 micromol L(-1) in a time-dose dependent manner. Treatment of SMMC-7721 cells with different concentrations of FQ16 for 24 h increased the percentage of the apoptosis cells obviously (P<0.05), the typical ladder DNA in apoptotic cells and a concomitant dissipation of the mitochondrial membrane potential. Compared with control group, FQ16 influenced obviously DNA topoisomerase II activity, stimulated DNA cleavage and inhibited DNA reunion mediated by topoisomerase II. In addition, FQ16 (3 - 7.39 micromol L(-1)) increased mRNA expression of Bax and protein expression of p53, Bax, caspase-9, caspase-3, separately, and induced cytosolic accumulation of activities caspase-9 and caspase-3, whereas the mRNA and protein expression of Bcl-2 decreased with no change of caspase-8. Therefore it can be concluded that the effects of inhibited topoisomerase II and mitochondrial-dependent pathways were involved in FQ16 induction of apoptosis of SMMC-7721 cells.

OBJECTIVE:To observe the effect of Compound glycyrrhizin on serum VEGF of patients with alopecia areata. METHODS:36 alopecia areata patients were selected as trial group and treated with Compound glycyrrhizin(50 mg/time,3 times/d) for 3 months. Double-antibody sandwich ELISA method was adopted to determine the content of VEGF in serum before and after treatment. 36 healthy volunteers were included in control group,and serum content of VEGF in control group was compared with trial group. RESULTS:Compared with control group,the serum content of VEGF was lower in trial group,with statistical signifi-cance(t=5.863,P<0.05). After treatment,the serum VEGF levels of trial group increased,with statistical significance(t=4.351, P<0.05);there was no statistical significance in VEGF levels between trial group and control group after treatment(t=1.678,P>0.05). CONCLUSIONS:Compound glycyrrhizin can cure alopecia areata through increasing serum level of VEGF.

Objective: To observe the effect of balancing yin-yang needling manipulation on post-stroke upper limb spasticity and changes of electromyography (EMG) after treatment.
Methods: A total of 60 eligible cases were randomly allocated into an observation group and a control group, 30 in each group. Based on routine medication, cases in the control group were treated with conventional needling manipulation, whereas cases in the observation group were treated with balancing yin-yang manipulation. After the courses of treatment were completed, the therapeutic efficacies were evaluated using modified Ashworth scale and clinical spasticity index (CSI), coupled with the integrated electromyography (IEMG) and root mean square (RMS) value of biceps in passive flexion of the elbow joint during isokinetic testing recorded with the surface EMG.
Results: The total effective rate in the observation group was 86.7%, versus 53.3% in the control group, showing a statistical significance (P < 0.05). After treatment, there was between-group statistical significance in severity of elbow spasticity (P<0.05); there were intra-group (P<0.01, P<0.05) and inter-group (P<0.05) statistical significances in CSI index;and there were intra-group (P<0.01, P<0.05) and inter-group (P<0.01, P<0.05) statistical significances in IEMG and RMS values.
Conclusion:Balancing yin-yang and conventional needling manipulations can both improve upper limb spasm and reduce CSI as well as IEMG and RMS values in stroke patients;however, balancing yin-yang manipulation is better than conventional manipulation in clinical effect.