OBJECTIVE To investigate the effect and mechanism of modified Lichong decoction （MLCD） on the apoptosis of transplanted tumor cells in nude mice. METHODS Human gastric cancer AGS cells were cultured， and a nude mice transplanted tumor model was established. The nude mice were divided into the model group and MLCD low-， medium- and high-dose groups （150， 300， 600 mg/kg）. They were given distilled water or the corresponding drug solution by gavage once daily for four consecutive weeks. The size of transplanted tumors in nude mice was measured every six days， and the tumor volume was calculated. After the medication， the nude mice were sacrificed， and the transplanted tumor tissues were isolated. The contents of lactate dehydrogenase （LDH） and reactive oxygen species （ROS） in the transplanted tumor tissues were detected， and the changes in mitochondrial membrane potential were assessed. The pathological morphological changes were observed. The enzymatic activities of caspase-3， caspase-8， and caspase-9， as well as protein expressions of Fas and FasL and mRNA expressions of caspase-3， caspase-8， caspase-9， Fas and FasL in the transplanted tumor tissues， were detected. RESULTS Compared with the model group， the volume of transplanted tumors in nude mice from all MLCD dose groups was reduced to varying degrees. The contents of LDH and ROS， as well as the enzymatic activities of caspase-3， caspase-8 and caspase-9， were significantly increased/enhanced. The mitochondrial membrane potential was significantly decreased. The protein expressions of Fas and FasL， and the mRNA expressions of caspase-3， caspase-8， caspase-9， Fas and FasL were significantly up-regulated. Most of these differences were statistically significant （ P ＜0.05 or P ＜0.01）. Pathological results showed that with increasing doses of MLCD， the cellular density in the transplanted tumor tissues gradually decreased， and typical morphological features of apoptosis， such as loosening and increasing fragmentation， became more prominent. CONCLUSIONS MLCD can induce apoptosis in transplanted tumor cells of nude mice， and its mechanism may be related to the activation of the Fas/FasL pathway and the caspase apoptotic pathway.

Objective:To compare the effects of Feishu(BL13)combined with Tianshu(ST25)and Feishu(BL13)alone on pulmonary function and tissue inflammation in asthma model rats. Methods:Forty-eight Sprague-Dawley rats were randomly divided into a normal group,a model group,a lung treatment group,and a lung-intestine treatment group according to the random number table method,with 12 rats in each group.Except for the normal group,rats in the other three groups were sensitized by ovalbumin followed by atomization stimulation to establish the asthma model.After the model was made successfully,rats in the lung treatment group were treated with acupuncture at bilateral Feishu(BL13)for 30 min;rats in the lung-intestine treatment group were treated with acupuncture at bilateral Feishu(BL13)and Tianshu(ST25)for 15 min,respectively,for 30 min in total.Acupuncture was performed once a day for 14 d.Rats in the other two groups did not receive any intervention.After intervention,the pulmonary function of each group of rats was measured by a pulmonary function tester.The histopathological changes stained by hematoxylin-eosin staining and the collagen deposition degree stained by Masson in lung tissue,as well as the inflammatory cells in the bronchoalveolar lavage fluid(BALF)stained by Wright's-Giemsa staining were observed under a light microscope;the levels of interleukin(IL)-4,IL-5,IL-13,IL-17,IL-25,IL-33,leukotrienes(LT),thymic stromal lymphopoietin(TSLP),and prostaglandin D2(PGD2)were detected by enzyme-linked immunosorbent assay. Results:Compared with the normal group,the peak expiratory flow(PEF),the dynamic lung compliance(Cdyn),the forced expiratory flow 25%(FEF25%),the forced expiratory volume in the first second/forced vital capacity(FEV1/FVC),and the maximum mid-expiratory flow(MMEF)in the model group were significantly decreased(P<0.01 or P<0.05);the lung resistance(RL),the collagen deposition,the IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 levels,and the neutrophil proportion in the BALF were increased significantly(P<0.01 or P<0.05).Compared with the model group,the FEF25%and FEV1/FVC in the pulmonary function were significantly increased(P<0.01,P<0.05),while the collagen fiber deposition and the levels of IL-4,IL-5,IL-13,IL-17,LT,TSLP,and PGD2 were significantly decreased(P<0.01 or P<0.05)in the lung treatment group;the PEF,FEF25%,and FEV1/FVC in pulmonary function were significantly increased(P<0.01 or P<0.05),while the RL and the collagen fiber deposition,as well as the neutrophil proportion and the levels of IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 in the BALF were significantly decreased(P<0.01 or P<0.05)in the lung-intestine treatment group.Compared with the lung treatment group,the IL-5 level in the rat lung tissue of the lung-intestine treatment group was decreased(P<0.05). Conclusion:Acupuncture in the lung treatment and the lung-intestine treatment groups improved pulmonary function,lung inflammation,and inflammatory cells in the BALF in asthma model rats;the lung-intestine treatment was better than lung treatment in improving the IL-5 level.The combination of Feishu(BL13)and Tianshu(ST25)was superior to Feishu(BL13)alone in the treatment of asthma in trend.

Objective To analyze the secular trends of pancreatic cancer incidence and mortality in Changning district of Shanghai from 1974 to 2013. Methods We calculated the age-standardized rates of incidence and mortality and the average annual percent changes for pancreatic cancer using Segi's world standard population and the data from Shanghai Cancer Registry. Age-period-cohort model was constructed to further assess the effect of age, diagnosis period and birth cohort on the secular trends of pancreatic cancer incidence and mortality. Results During 1974-2013, the age-standardized incidence and mortality rates were 6.49/10⁵ and 6.01/10⁵ in male, 4.83/10⁵ and 4.57/10⁵ in female, respectively. The age-standardized incidence was increased by 0.8% per year in male during past 40 years, while there was no change in mortality. The age-standardized incidence and mortality rates were increased by 1.6% and 1.3% per year in female. After adjusting the effects of diagnosis period and birth cohort, the incidence and mortality rates of pancreatic cancer increased by about 11% every 5 years older in both male and female. Diagnosis period and birth cohort had no statistical impact on the incidence and mortality of pancreatic cancer. Conclusion The age-standardized incidence of pancreatic cancer shows significantly rising trends during 1974-2013 in both male and female in Changning district of Shanghai, as well as the age-standardized mortality in female. The incidence and mortality rates also increase with age.

Objective To investigate the effects of adoptive reinfusion of regulatory T cell (Treg) on the recovery of islet function and graft survival time after islet allograft transplantation. Methods The diabetic model was established using C57BL/6 mice as recipients, and Balb/c mice were chosen as donors for islet allografts transplantation beneath the renal capsule. The recipient mice were divided into 3 groups and 3 mice in each group according to different processing Methods: Treg experiment group (Treg group, 1×10⁶ Treg cells were injected via tail vein at 1 d before operation), positive control group [sirolimus (SRL) group, SRL at a dose of 300 μg/(kg·d) was intragastrically given every day from 1 d before operation] and blank control group (control group, an equivalent volume of normal saline was intragastrically given every day from 1 d before operation). Enzyme-linked immune absorbent assay (ELISA) was used to detect the changes of blood glucose and C-peptide in mice within 14 days after transplantation. In vivo imaging technique was used to dynamically monitor the survival of mice within 14 days after transplantation. Results In each group, the blood glucose levels at postoperative 3 d were significantly decreased compared with those before transplantation (all P < 0.001). At postoperative 1 d, the C-peptide levels showed an explosive rise to varying degree in each group. At postoperative 3 d, the C-peptide levels in each group were significantly higher than that before operation (all P < 0.001). At the end of the observation period at 14 d after operation, the C-peptide levels in the SRL and Treg groups were (427±50) pmol/L and (833±57) pmol/L, relatively higher than that in the control group. But the blood glucose levels were (14.5±0.5) mmol/L and (12.1±0.6) mmol/L, significantly lower than that in the control group (all P < 0.001). Compared with the SRL group, the explosive release amount of C-peptide was significantly lower, the declining trend was more remarkably stable, and the C-peptide level was considerably higher in the Treg group at the end of the observation period (all P < 0.001). At postoperative 14 d, the grafts were almost completely apoptotic in the control group, over 50% of the grafts survived in the SRL group, and over 80% of the grafts survived in the Treg group. Conclusions Adoptive reinfusion of Treg cells can effectively protect islet grafts, prolong the survival time of grafts, and maintain the normal levels of blood glucose and C-peptide in the recipient mice.

Ten compounds were isolated and purified from the ethyl acetate fraction of Houttuynia cordata by silica gel and ODS column chromatographies. The chemical structures of the compounds were identified on the basis of physicochemical properties as well as spectral data. These isolated compounds were elucidated as sequinoside L(1), a new phenolic glycoside, together with nine known compounds, including sequinoside K(2), methyl chlorogenate(3), chlorogenic acid(4), hydroquinone(5), vanillic acid(6), hyperin(7), isoquercitrin(8), quercetin(9), and quercitrin(10). In addition, the anti-inflammatory activities of compounds 1-10 were evaluated in LPS-induced RAW264. 7 macrophages. Among them, compounds 1, 2, 6 and 9 showed potent anti-inflammatory activity which was similar to that of positive control dexamethasone.

Objective To investigate the prognosis of patients underwent liver transplantation (LT) for end-stage autoimmune liver disease (AILD). Methods Clinical data of 48 patients with end-stage AILD undergoing LT from May 1996 to April 2013 in Affiliated Changzheng Hospital of the Second Military Medical University were analyzed retrospectively. The postoperative cumulative survival rates of the recipients were calculated,and the cause of death was analyzed. The postoperative rejections,new-onset viral hepatitis and AILD recurrence were analyzed. Results In 48 AILD recipients,38 cases survived and the postoperative 5-year cumulative survival rate was 76%. Causes of death for the 10 dead cases were multiple organ failure, liver graft failure,sepsis,pulmonary infection,hemorrhage,hepatic artery embolization and renal failure. In 48 AILD recipients,9 cases (19%) suffered acute rejection after operation,3 cases suffered new-onset hepatitis B infection in 1-2 years after operation,2 recipients suffered primary disease (primary biliary cirrhosis)recurrence 2 years after operation and all survived for a long term after positive treatments.Conclusions Most liver transplant recipients with end-stage AILD can obtain a long-term survival. Attentions should be paid on the immunosuppressive regimens in early period after LT,prevention of infection,rejection and postoperative new-onset viral hepatitis,timely diagnosis of primary disease recurrence.

BACKGROUND:Recently, scholars al over the world have made numerous studies on three-dimensional visualization model of reconstructing maxil ofacial bone tissue, skin, and muscle in the ordinary PC. However, few studies concerned the three-dimensional visualization model of constructing maxil ofacial bone tissue, skin, muscle and blood vessels. OBJECTIVE:To establish a three-dimensional visualization model of the complete maxil ofacial region using an ordinary PC based on spiral CT, MRI data, and three-dimensional reconstruction software. METHODS:One healthy male adult was chosen as the object in this study. CT and MRI were performed respectively to acquire the DICOM standardized image of the sample. Al CT and MRI images were imported to Mimics to construct three-dimensional visualization models of the bones in the maxil ofacial region, some masseter muscles, trigeminal cistern, internal carotid artery and internal jugular vein. Three-dimensional model of the bones in the maxil ofacial region was chosen as the reference. Models of muscles, blood vessels and trigeminal cistern established by MRI images were imported, and registration in space of three-dimensional model was performed. At last, three-dimensional visualization model of the complete maxil ofacial region was acquired. RESULTS AND CONCLUSION:The integrated three-dimensional models of the bones, muscles, skin, trigeminal cistern and blood vessels in the maxil ofacial region established accurately reflected the complex anatomic structure of maxil ofacial region, provided reliable anatomic data for clinical diagnosis and laid a good foundation for sham operations in the future.

10.3969/j.issn.2095-4344.2013.24.002

Objective To investigate the protective effect of a COX inhibitor,flurbiprofen (Flurb) on hepatic ischemia/reperfusion (IR) injury in rats and the action mechanism.Method C57BL/6 mice were randomized into sham,IR and Flurb (4 different doses) groups.The model of segmental (70％) warm hepatic ischemia was established in IR and Flurb groups.Flurbiprofen of different doses (5,7.5,10 and 15 mg/kg) was injected via the tail vein 20 min before ischemia.At different time points after reperfusion,liver cell necrosis and apoptosis were evaluated by HE and TUNEL staining.The COX and inflammatory cytokine gene expression was detected by using realtime PCR.Liver mitochondria were separated and mitochondrial permeability transition (MPT) pore sensitivity was examined by using swelling assay and fluorescence spectrophotometry assay.Result In flurbiprofen groups of different doses,the serum AST and ALT levels were significantly decreased at 6 h after reperfusion as compared with IR group.Moreover,10 mg/kg Flurb pretreatment significantly inhibited the mitochondrial permeability transition (MPT) pore opening,and thus alleviated liver cell damage and prevented mitochondria-related cell death and apoptosis by inhibiting COX-2 and inflammatory factor genes expression such as IL-1β,IL-6 and TNF-α.Conclusion Flurbiprofen protects mice from hepatic I/R injury possibly by inhibiting mitochondrial permeability transition and IL-1β,IL-6 and TNF-α expression,which may provide experimental evidence for clinical use of flurbiprofen to protect liver function in surgical settings other than its conventional use for pain relief.

BACKGROUND:Studies have shown that low-intensity and short-time vibration with a certain frequency can reduce the absorption of bone tissue and increase the quantity and quality of bone through promoting the proliferation and differentiation of osteoblasts.
OBJECTIVE:To investigate the effects of different frequencies of vibration strains on cycle, proliferation and differentiation potency of RAW264.7 cel s cultured in vitro.
METHODS:Passage 6 RAW264.7 cel s in good conditions were randomly divided into six groups, and each group was induced cultured with Dulbecco’s modified Eagle’s medium containing receptor activator of nuclear factor kappa B ligand. The final concentration of receptor activator of nuclear factor kappa B ligand was adjusted to 50μg/L, and then kept without changes. The non-loading group did not loaded with vibration strain, and the other five groups were loaded with 3-10 Hz, 15-35 Hz, 35-45 Hz, 50-70 Hz and 70-90 Hz vibration strains on the RAW264.7 cel s respectively. The other vibration parameters were consistent;the vibration time was 15 min/time with the vibration intensity of 0.3 g, twice per day. The cel cycle and cel proliferation were detected at 3 and 6 days after loading of vibration strains.
RESULTS AND CONCLUSION:After composite vibration loading for 6 days, the cel cycle phase in the vibration groups was changed to some extent when compared with the non-loading group. Compared with non-loading group, the cel number in the G 1 phase of the vibration group was significantly increased (P<0.01);the cel number in the S phase and G 2+M phase of the vibration group was significantly decreased (P<0.01);the vibration parameters in the vibration group were significantly decreased (P<0.01). The results indicate that different frequencies of vibration strains can affect the cel cycle and proliferation, and can inhibit the proliferation and differentiation of osteoblasts.