AIM:To explore the protective effects of berberine (Ber) on islet beta cells and related possible mechanisms.METHODS:The injury of INS-1 cells was induced by treatment with alloxan ( Axn) .Berverine was then given at serial concentrations.The cells were divided into control group ( Con) , injury group ( Axn) , low-dose berberine group (LBer), medium-dose berberine group (MBer) and high-dose berberine group (HBer).Flow cytometry was em-ployed to detect the apoptosis.The activation of PTEN/PI3K/Akt and HNF-1α/PDX-1 pathways and the protein level of cleaved caspase-3 were evaluated by Western blotting.The insulin releases under normal or high-glucose stimulation were measured by ELISA.RESULTS:Compared with Con group, the apoptotic rate increased significantly in Axn group.Ber-berine treatment reduced the apoptotic rate in LBer group, MBer group and HBer group in a concentration-dependent man-ner.Compared with Con group, the protein levels of PTEN and cleaved caspase-3 increased, while PI3K and phosphoryla-tion of Akt decreased significantly in Axn group.However, this effect was reversed by berberine in a concentration-depend-ent manner.Compared with Con group, the activation of HNF-1α/PDX-1 signaling pathway was inhibited in Axn group but recovered by berberine administration.The abilities of releasing insulin under normal or high-glucose stimulation were im-paired in Axn group but recovered by berberine treatment in LBer group, MBer group and HBer group in a concentration-dependent manner.CONCLUSION:Berberine shows protective effects against alloxan-induced damage in beta cells by in-hibiting apoptosis and recovering insulin secretion, thus attenuating the activation of PTEN/PI3K/Akt and HNF-1α/PDX-1 signaling pathways.

Objective To detect estrogen receptor and DNA index in breast cancer by flow cytometry. Methods 32 cases of fresh specimens of breast cancer resected by operation were used in this study. Single cells suspension was prepared from those specimens, and estrogen receptor expression was analyzed by flow cytometry. At the same time, the status of ploid and S phase cell ratio were determined. Results Flow cytometry analysis could measure the expression level of estrogen receptor in the fresh breast cancer specimens, which was more sensitive and needed less volume of tumor tissue compared with immuohistochemical method. The information on the status of ploid and S phase cell ratio were simultaneously obtained. Conclusion The detection of estrogen receptor in breast cancers by flow cytometry was more helpful for evaluating prognosis and selecting treatment.

Objective To investigate the effects of sulforaphane (SFN) on the proliferation of prostate cancer DU145 cells and hypoxia-inducible factor-1α(HIF-lα) expression,and to explore the possibility of SFN in the prevention of prostate cancers.Methods The DU145 cells were treated with different doses and different time of SFN.The cell proliferation viability were measured by 3-(4,5)-dimethylthiahiazo (-zy1)-3,3,5-di-phenytetrazo`liumromide (MTT) proliferation and cytotoxicity assay kit.The hypoxia tumor microenvironment was stimulated by pretreating cells with 1％ O2,HIF-lα expression and vascular endothelial growth factor (VEGF) activitity were detected by Western-blot and ELISA,respectively.Results SFN had an inhibitory effect on the proliferation of human prostate cancer DU145 cells in a dose-and time-dependent manner.The cell proliferation inhibition rate exceeded 50％ in more than 20 μ mol/L SFN-treated groups after 48 hours.SFN could inhibit the expression of HIF-1α induced by 1％ O2 and VEGF activity.10 μmol/L SFN showed inhibitory effect on the expression of HIF-1α after 24 hours.20 μ mol/L SFN showed a significant inhibitory effect on the expression of HIF-1α after 12 hours.And peroxidase had synergistic inhibition on HIF-1α.Conclusions SFN can inhibit the proliferation of DU145 prostate cancer cells and HIF-lα expression.SFN can be used as a potential chemotherapeutic agent against human prostate cancer.

Since the HLA system is one of the most complex human genetic polym- orphisms,its application in forensic medicine included disputed paternity and criminal identification,have been fairly recognized. The present paper reported the results of our study about the HLA typing in human blood stain,serum and saliva,it was concluded that:(1).The existed strong anti-complementary activity in human blood stain when the amount of complement used in microlym-phocytotoxicity inhibition test(MLIT) was incresed to 10?l,it was found that the results of HLA-All,-B 5 typing in bloodstains were all correct,and the detectable period was at least 90 days; (2).The soluble HLA-A antigens in human serum could reliable detected with MLIT;(3).The soluble HLA-A antigens were also present in the human siliva.

OBJECTIVE:To investigate the effects of Wenxin granule on the related indexes of senile patients with primary hy-pertension and acute left cardiac failure. METHODS:160 senile patients with primary hypertension and acute left cardiac failure were randomly divided into control group(80 cases)and observation group(80 cases). Control group received 12.5 mg captopril, orally,twice to 3 times a day (if patients had taken plenty of diuretic drugs,in low sodium and/or low blood volume,patients with normal or low blood pressure,the initial dose was 6.25 mg,3 times a day). Observation group additionally given 9 g of Wenxin granule,3 times a day. The treatment course for both groups was 4 weeks. SBP,DBP,PP,SDANN,SDNN-Index,RMS-SD,PNN50,improvement of cardiac functions before and after treatment,and incidence of adverse reactions in 2 groups were ob-served. RESULTS:After treatment,the SBP,DBP and PP in 2 groups were significantly lower than before,and observation group was lower than control group,SDNN,SDANN,SDNN-Index、RMSSD and PNN 50 were significantly higher than before,and ob-servation group was higher than control group,the differences were statistically significant(P<0.05). And there was no significant difference in the incidence of adverse reactions(P>0.05). CONCLUSIONS:Based on conventional treatment,wenxin granule can significantly reduce the bloop pressure of senile patients with primary hypertension and acute left cardiac failure,improve cardiac functions and inhibit autonomic disorders,with good safety.

Objective To investigate effect of mild hypothermia on changes of somatosensory evoked potential (SEP) and synaptophysin mRNA level after traumatic brain injury (TBI) and determine hypothermia-induced neuroprotection.Methods Forty-five SD rats were allocated into mild hypothermia group,TBI group and sham operation group with 15 rats per group according to the random number table.Left-side fluid percussion impact was performed to induce models of TBI.Rats were exposed to hypothermia environment (32-35℃) for 6 hours in mild hypothermia group after TBI.Rats in sham operation group were treated by only drilling on left side of the head,rather than hitting.To evaluate function outcome,modified neurological severity score (mNSS),SEP and synaptophysin mRNA level were measured at 6 hours,24 hours and 7 days postinjury.Results The mNSS in mild hypothermia group lowered compared with TBI group,especially at 24 hours and 7 days (P ＜ 0.05).SEP in mild hypothermia group was significantly shortened at 6 and 24 hours compared with TBI group (P ＜ 0.05),but SEP revealed no significant difference among the 3 groups at 7 days (P ＞ 0.05).Level of synaptophysin mRNA in mild hypothermia group increased at 6 hours postinjury compared with TBI group [(0.08 ± 0.02) vs (0.12 ±0.04)],with further increase at 7 days postinjury[(0.06 ± 0.01) vs (0.33 ± 0.10)] (P ＜0.05).Conclusion The shortage of nerve conduction time of the injured side and promotion of nerve regeneration suggest the neuroprotective role of mild hypothermia following TBI.

Objective:To compare the diagnostic value of strain ratio correction in ultrasound elasticity imaging and ultrasound-guided percutaneous biopsy for the diagnosis of breast imaging reporting and data system (BI-RADS) 4 breast mass. Methods:From January 2014 to June 2016 in the Affiliated Hospital of Chengde Medical College, 120 patients with breast tumor resection and preoperative BI-RADS4 of all the masses were included in this study. The diagnostic value of contrast ultrasound elastography strain ratio correction and biopsy in ultrasound-guided percutaneous biopsy was evaluated with pathology as the gold standard. Results:In the 120 cases of grade BI-RADS4 mass by ultrasound elastography after correction, 46 cases were graded BI-RADS4 without changing;59 cases were downgraded to BI-RADS3, and 15 cases were upgraded to BI-RADS5. The pathology, ultrasonic elastography strain ratio correction, biopsy diagnostic sensitivity, specificity, and accuracy were 90.7%, 81.8%, 85.5%, 88.8%, 98.5%, and 95.0%, respectively. A significant difference was found between ultrasonic elastography strain ratio correction and ultrasound-guided biopsy for the diagnosis of breast malignant mass (P<0.05), whereas no significant difference was found for the diagnosis of benign mass (P>0.05). Conclusion:Ultrasound-guided puncture biopsy is highly valuable in qualitative diagnosis of BI-RADS4 breast mass. After the pathological diagnosis, ultrasound elastography strain ratio has certain clinical value in BI-RADS4 guided biopsy and correction of mass classification.

OBJECTIVE To analyze the sequence of IMP-4 metallo-?-lactamases(MBLs) encoding gene from clinical isolates of multiple-drug-resistant Klebsiella oxytoca strains and attempt to know the integrons composing the drug resistance gene box.METHODS The antibiotic sensitivity test of multi-resistant K.oxytoca strains was done according to Kirby-Bauer method of CLSI 2005,and the double disk synergy test and Etest were for detecting their MBLs.The Class 1 integrons were detected by PCR.The purified amplicons of Class l integrons were sequenced.The type and order of gene cassettes in integrons were analyzed by searching GenBank.RESULTS The K.oxytoca was resistant to carbapenems,the third-generation cephalosporins,cefoxitin,quinolones,cefoperazone/sulbactam,sulfamethoxazole/trimethoprim,amoxicillin/clavulanate,ticacillin/clavulanate,piperacillin,cefepime,rifampicin and piperacillin/tazobatam,only susceptible to amikacin and polymyxin B.The IMP-4 metallo-?-lactamases,aadA1,AmpC,CTX-M-14,qacE△1-sull and intI1 were positive.CONCLUSIONS Integrons are important molecular mechanism in the development of multidrug resistance.There are resistance gene boxes in them.

Objective To study the inhibition of anti-PDGF on proliferation of cultured human retinal pigment epithelial cells(hRPE) in vitro.Methods hRPE were cultivated and were exposed to different concentrations of anti-PDGF(0,1?10-6,5?10-6,1?10-5,5?10-5 and 1?10-4 mg?L-1) respectively .Growth curves were measured with cell counting and the vitalities of cells were examined by percentage of vital cells and total cells.Using MTT staining colorimetric to measure the inhibitory rate.The changes of cell cycle of hRPE were collected and their growth were detected with FCM analysis and the morphological changes of cells were observed by light microscope and electron microscope.Results Anti-PDGF of 1?10-6mg?L-1 stimulated hRPE proliferation slightly.AntiPDGF at dosages ranging from 5?10-6mg?L-1 to 1?10-4mg?L-1 inhibited cell proliferation effectively in a dose-dependent and time-dependent manner(P

BACKGROUND: Caspase-3 is well recognized as the key caspase carrying out apoptosis in animal and human brain. To date, a few studies revealed the expression of caspase-3 protein in brains of normal persons and Alzheimer patients but data obtained from rodents exhibited much discrepancy.OBJECTIVE: To investigate the different expression patterns of caspase-3in rodent and monkey brain, and the different expression of caspase-3 in different brain regions and during aging in monkeys.DESIGN: Parallel comparison between means of single variable.SETTING: Institute of Hepatobiliary Surgery, Chinese PLA General Hospital and Department of Anatomy, the Chinese University of Hong Kong.MATERIALS: The experiment was carried out from August, 2003 to February, 2005 in Institute of Hepatobiliary Surgery, Chinese PLA General Hospital and Department of Anatomy, the Chinese University of Hong Kong. Sprague Dawley rats, ICR mice and senescence-accelerated mice (SAM) with ages ranging from postnatal 2, 12, 24 to 48 weeks(n=5 for each age group of different rodents) were included in the present study. All of these animals were supplied by Laboratory Animal Services Center, the hinese University of Hong Kong. Totally 8 rhesus monkeys aged 4 years (n=4) or 20 years (n=4) were selected from the Laboratory Animal Center in Chinese PLA General Hospital [SCXK-(Beijing)2003-002]. Both ro dents and monkeys were female and were raised under standard conditions without any experimental interventions. METHODS: ①Brain tissue samples were taken freshly from both rodents and monkeys and made into homogenate. The expression of caspase-3 pro tein in brains of both rodents and monkeys was investigated with im munoblot. ② The expression levels in monkey brains were exhibited quantitatively with the same method in three brain regions, such as the frontal cortex, hippocampus and cerebellar cortex, for the two age-groups. In vivo distribution patterns of caspase-3-immunoreactive cells were further presented in 3 brain regions of monkeys through immunohistochemistry. MAIN OUTCOME MEATURES: ①Detection of caspase-3 protein with immunoblot in the brain of rodents and monkeys; ② Distribution patterns of caspase-3-immunoreactive cells in 3 brain regions of monkeys. RESULTS: ① Result of detection with immunoblot: The same pattern of caspase-3 protein expression in brain of three kinds of 2-week-old rodents. But the expression was not seen in any other brains of older ages. Caspase 3 was expressed in a relatively high level inboth adult and aged monkey brains, and the amount did not attain to the level in 2-week-old rodents. Caspase-3 Was expressed in the pattern of zymogen (Mr 32 000). The ex pressions of caspase-3 in brains of monkey were not different in ages and brain regions. ②Result of Immunohistochemistry: It was showed that most neurons in the frontal cortex lack detectable caspase-3 immunoreactivity, whereas low to moderate caspase-3 immunostaining be found mainly in pyramidal cells in CA1, CA3 and CA4 subfields of hippocampus. And in the cerebellum, a small number of Purkinje cells were strongly stained in their cytosol and dendrites. Age-related expression pattern of caspase-3 were not found except that in the motor cortex of aged monkeys in which there were a limited number of large pyramidal cells in layer Ⅴ that were strongly stained with caspase-3 antibody.③ Immunoblot procedure revealed that the caspase-3 protein expressed in monkey brains is in the form of zymogen (Mr 32 000) and there is no significant difference in caspase-3 expression level as a function of either brain region or age of animals.CONCLUSION: Unlike rodents in which caspase-3 protein rapidly drops to an undetectable level since animals grow up, the primate expresses caspase-3 constitutively in brain until the late period of lifetime. But there are no significant brain region- or age-related differences in the protein levels in monkey brain.