Objective To evaluate the association between cytomegalovirus (HCMV) infection and atherosclerosis (AS) and explore the possible role of endothelin (ET) and serum tumor necrosis factor(TNF ?) in the pathogenesis of AS in diabetic patients and the relationship between HCMV infection and concentrations of ET and TNF ?. Methods Blood samples were collected from 21 patients with AS of type 2 DM, 47 patients with non AS of type 2 DM and 20 controls. Special antibodies to HCMV (IgM,IgG) were measured by enzyme linked immunosorbent assay (ELISA) . The concentrations of serum TNF ? and plasma ET were determined by RIA. Results (1) Type 2 diabetic patients, especially those with established AS, had a higher prevalence of active infection of HCMV. (2) The levels of plasma ET and serum TNF ? in type 2 diabetic patients were higher than those in normal people. In the pathogenesis of AS in diabetic patients, the ET played more important role than the TNF ?.(3) There was a significantly positive correlation between anti IgM to HCMV and the level of plasma ET in type 2 DM patients. It suggested that the injury of endothelium cell was related to the active infection of HCMV. Conclusion The study suggests that HCMV infection may be important in the pathogenesis of AS in type 2 diabetic patients and ET possibly plays an important role on the pathological course of AS.

Background and purpose:Oxaliplatin is a new cytotoxic platinum compound widely used in antineoplastic treatments.Peripheral neuropathy characterized by allodynia remains the most common way to limit the usage of oxaliplatin.Oxaliplatin-associated neuropathic pain is often resistant to standard analgesics.The effects of antidepressant agents such as desipramine and fluoxetine on chemotherapy-induced neuropathic pain were investigated so as to provide experimental evidence for clinical treatment.Methods:A single injection of oxaliplatin(30 mg/kg)intraperitoneal was injected into a test subject,a mouse that had chronic neuropathic pain.Using the von Frey filament as a touch stimulator,the mechanical withdrawal threshold(MWT)was measured when observing allodynia.The MWT was measured before and 1 h after the administration of desipramin and fluoxetine.Results:Desipramine and fluoxetine both have the potential to increase the MWT in mice with oxaliplatin-induced neuropathic pain.Pretreatment with antagonists such as an opioid receptor like naloxone could deepen their effects.Furthermore,when desipramine is combined with an opioid analgesic as buprenorphine,it causes an augmentation in the MWT.Conclusion:Antidepressants desipramine and fluoxetine antagonize oxaliplatin-induced neuropathic pain by inhibiting allodynia.Furthermore,the tricyclic antidepressing agent desipramine could enhance the effects of buprenorphine in subjects with oxaliplatin-induced pain,suggesting a synergistic effect for opioid analgesic.

Objective To explore neurobehavioral changes in rats with recurrent seizures and the prevention effect of melatonin.Methods 6-day-old (P6) SD rats were randomly divided into four groups of 24 (n =6):the control group (CONT),melatonin per se group (MEL),recurrent neonatal seizure group (RS) and melatonin administration prior to RS group (RS + MEL).Rats in RS group were subjected to 5 seizures with flurothyl during the first 14 days of life.In RS + MEL group,melatonin was injected at 8:00 before seizures were induced.Neurobehavioral tests including Plane righting experiment,Cliff avoidance test,the grip-strength test and negative geotaxis test were implemented on P24,while open field test on P35.Results (1) Plane righting experiment:the time of plane righting in RS group ((0.33 ± 0.51)s) was significantly shorter than that in the CONT group ((1.17 ± 0.40) s) and RS + MEL group ((0.50 ± 0.54) s) (P ＜ 0.05).(2) Cliff avoidance test:the time of cliff avoidance in RS group ((16.00 ± 6.32) s) was significantly longer than that in CONT group ((4.00 ± 2.60) s)(P ＜ 0.01),while the time of cliff avoidance in RS + MEL group ((7.67 ± 3.26) s) was shorter than that in the RS group (P ＜ 0.05).(3) The grip-strength test:compared with CONT group ((49.50 ± 28.96) s),the time needed to hold on wire in RS group((11.67 ± 7.58)s)was significantly shorter (P ＜ 0.05) and longer in RS+ MEL group ((24.83 ± 6.61) s) (P ＜ 0.05).(4) Negative geotaxis test:the time for rats to turn 180° upward in RS group((7.67 ± 1.36) s) was longer than that in the CONT group ((4.50 ± 2.66) s) and RS + MEL group ((6.17 ± 0.75) s) (P ＜ 0.05).(5) Open field test:the time for rats to begin to run in the RS group ((8.17 ± 3.86) s) was longer than that in the CONT group ((3.00 ± 1.41) s) (P ＜ 0.05).Conclusion The neurobehaviors are damaged following flurothyl-induced recurrent neonatal seizures,and melatonin can reduce the neurobehavioral injury.

Objective To investigate dynamic expressions of cortex clusterin (CLU) and intervention effect of ketogenic diet (KD) on neonatal rats with recurrent seizures.Methods Thirty-six-8-day postnatal SD rats were randomly divided into 3 groups:normal control group (NS + ND group,n =12),and the recurrent-seizure and normal diet group (RS + ND group,n =12),and the recurrent-seizure and KD group (RS + KD group,n =12).From 9 d,rats in RS + ND group and RS + KD group were subjected to recurrent seizures induced by volatile flurothyl 30 min each day for consecutive 8 days.Rats in NS + ND group were placed into the container for an equal amount of time to their counterpart without exposure to flurothyl.Scores on neurological behaviors at 35 days postnatally were examined.CLU protein levels in cerebral cortex were determined by Western blot at 58 days postnatally.Results Neurodevelopmental indicators analysis:in the plane righting experiment,there were significant differences between NS + ND group [(1.03 ± 0.54) s],R S + KD group [(0.89 ± 0.16) s] and RS + ND group [(0.64 ± 0.30) s] about the time of plane righting (all P ＜ 0.05) ; in the negative geotaxis reaction experiment,the rats of NS + ND group [(1.92 ± 0.90) s],and RS + KD group [(5.17 ± 0.72) s] about the time of negative geotaxis reaction were significantly different compared with RS + ND gouup [(7.33 ± 0.65) s] (all P ＜ 0.01).In the cliff avoidance test,there were significant differences between NS + ND group,R S + KD group [(4.33 ± 2.54) s,(8.75 ± 2.26) s] and R S + ND group [(16.58 ± 4.25) s] about the time of cliff avoidance (all P ＜ 0.01).Western blot showed that the expression of CLU in cerebral cortex of the RS + ND group [(2.24 ± 0.53) s] was obviously increased compared with NS + ND group [(1.44 ± 0.11) s] (P ＜0.01),and there also had significant difference between RS + KD group [(1.56 ±0.24) s] and RS + ND group (P ＜ 0.05).Conclusions It shows that the up-regulated expression of CLU in cerebral cortex may be associated with recurrent neonatal seizure-induced brain damage,while KD may protect them from recurrent neonatal seizure-induced brain damage by down-regulating expression of CLU.

The COI gene as DNA barcode was used to identify the Manis pentadactyla and its adulterants in order to provide a scientific basis for the molecular identification of M. pentadactyla. Genomic DNA was extracted from experimental samples using the DNA extraction kit. The COI genes were amplified using polymerase chain reaction (PCR) and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. The neighbor-joining (NJ) tree was constructed by MEGA 6.0. The results indicated that COI sequences were successfully amplified and NJ trees results indicated that M. pentadactyla and its adulterants can be easily identification. Therefore, the COI gene is an efficient barcode for identification of M. pentadactyla and its adulterants,which will provide a new technique for the market supervision.
Animals ; Cattle ; DNA Barcoding, Taxonomic ; methods ; Drug Contamination ; prevention & control ; Electron Transport Complex IV ; genetics ; Mammals ; classification ; genetics ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Sheep ; Swine

Objective: To study the effects of endostatin and doxycycline on microcirculatory patterns in the melanoma transplant model of the chorioallantoic membrane of the chicken embryo and the experimental evidences for melanoma therapy thereof. Methods: Endostatin was dripped on the chorioallantoic membrane after melanoma was transplanted 3 days (A group) and 5 days (B group). Doxycycline was dripped on the chorioallantoic membrane after melanoma was transplanted 3 days (C group) and 5 days (D group). The PBS solution was dripped on the chorioallantoic membrane in control group (E group). The hematoxylin-eosin（HE）staining and immunohistochemical staining were performed to observe microcirculation patterns in sections. Results: Compared with the E group, the area of endothelium-dependent vessels were significantly decreased in A group and B group (P ＜ 0.01). But the areas of vasculogenic mimicry were bigger in the two groups(P ＜ 0.05). Compared with E group, the areas of endothelium-dependent vessels and vasculogenic mimicry were significantly decreased in D group(P ＜ 0.05). Conclusion: Endostatin can inhibit angiogenesis. But it has no effect on vasculogenic mimicry in the transplant melanoma. And Doxycycline can inhibit vasculogenic mimicry and angiogenesis in the transplant melanoma. The results provided experimental evidences for melanoma therapy.

Objective To investigate the intervention effect of ketogenic diet (KD) on neurobehavioral demages after flurothyl-induced recurrent neonatal seizures in rats and on the expression of ApoE.Methods Postnatal day 8 (P8) SD rats (quantity:48) were randomly divided into two groups:the non-seizure group (NS group,n =24) and the recurrent-seizure group (RS group,n =24).From P9,rats in RS group were subjected to recurrent seizures induced by volatile flurothyl 30 min each day for consecutive 8 days.While rats in NS group were placed into the container for an equal amount of time to their counterpart without exposuring to flurothyl.At P28,each group was divided into two groups again:non-seizure and normal diet group(NS + ND group,n =12),non-seizure and ketogenic diet group(NS + KD group,n =12),recurrent-seizure and normal diet group (RS + ND group,n =12),recurrent-seizure and ketogenic diet group(RS + KD group,n =12).At P42,neurodevelopmental indicators were monitored.ApoE protein levels in cerebral cortex were determined by western blot at P58.Results Neurodevelopmental indicators were analyzed at P42:in the plane righting experiment,the rats of group NS + ND (1.0 ±0.14) about the time of plane righting was significant different comparing with group RS + ND ((0.75 ±0.32) s) (P ＜ 0.05).There was no significant difference between group RS + KD and group RS + ND about the time of plane righting(P＞ 0.05).In the negative geotaxis reaction experiment,the rats of groups NS + KD and RS + ND((3.17 ± 0.58)s,(6.75 ± 0.75)s) about the time of negative geotaxis reaction were significant different comparing with group NS + ND ((1.58 ±0.52)s) (P＜0.05).Compared with group RS + ND,the group RS + KD in the time of negative geotaxis reaction was obviously shortened (P ＜ 0.05).In the cliff avoidance test,there were significant differences between group NS + ND、RS + KD ((5.75 ± 2.90) s,(9.50 ± 4.36) s) and group RS + ND ((14.00 ± 4.79) s) about the time of cliff avoidance (P ＜ 0.05).In western blot,the expression of ApoE in cerebral cortex in the RS + ND group (1.26 ± 0.30) was obviously increased compared with group NS + ND (0.78 ±0.12) (P＜0.05),and there had also significant difference between group RS + KD (0.89 ±0.10) and group RS + ND (P ＜ 0.05).Conclusion The neuroprotective effects of ketogenic diet on recurrent neonatal seizure-induced neurobehavioral demages may be associated with the reduction of ApoE in cerebral cortex.

Objective To compare the effect of routine ketogenic diet and every other day ketogenic diet on neurobehavioral damage induced by recurrent seizures in neonatal rats.Methods 48 postnatal day 8 (P8) SD rats were randomly divided into four groups with 12 in each group:the control group (CONT),the recurrent seizure group(RS+ND),recurrent seizure + routine ketogenic diet group(RS+KD) and recurrent seizure+ every other day ketogenic diet group(RS+KOD).The recurrent seizures model was induced by flurothyl at p9 and last for 8 days.After a day of fasting the postnatal 28 day rats were placed on either ordinary or ketogenic diet according to packet design.Plane righting experiment,cliff avoidance test and negative geotaxis test were used to assess the neurobehavioral performance at p35.Results (1) Plane righting experiment:the plane fighting time of RS+ ND group ((0.17±0.39) s) was significantly shorter than that of NS+ND group ((0.67 ±0.49) s) (P＜0.05) ; and the plane righting time of RS+KD group((0.58±0.52) s) was significantly longer than that of RS+ND group ((0.17±0.39) s) (P＜0.05).There was no statistical significance between RS+KOD group((0.17±0.39) s) and RS+ND group ((0.17±0.39) s) (P＞0.05).(2) Cliff avoidance test:the cliff avoidance time of RS+ND group ((12.58±4.83) s)was significantly longer than that of NS+ND group ((1.92±0.90) s),RS+KD group((3.33± 1.50)s) and RS+ KOD group (P＜0.05) ;and the cliff avoidance time of RS+KOD group((5.58± 1.93)s) was significant longer than that of RS+KD group ((3.33± 1.50) s) (P＜0.05).(3) Negative geotaxis test:the negative geotaxis time of RS+NDgroup((3.17±1.70)s) was significantly longer than that of NS+ND group((1.42±0.67) s) and RS+KD group ((1.42±0.52)s) (P＜0.05).There was no statistical significance between RS+KOD group and RS+ND group(P＞0.05).Conclusion The ketogenic diet can improve neurobehavioral damage caused by flurothyl-induced recurrent seizures in neonatal rats.The every other day KD group shows a weak intervention effect comparing with the routine KD group.

Objective To investigate the effect of L-selectin in experimental allergic encephalomyelitis (EAE) of Wistar rats.Methods The rats were randomly divided into four groups;the normal group,the CFA group, the LMS group and the model group;The animal models were established in rats by immunization with myelin basic protein of spinal cord of guinea pig and complete Freund's adjuvant(CFA).The symptom of EAE was observed; pathological feature and myelin of brain and spinal were detected with HE stain and Loyez's stain respectively.The number of positive vessels of L-selectin expression was detected by immunohistochemistry.Results 100% experimental Wistar rats treated with MBP and levamisole developed EAE,but none of the other groups.The number of positive vessels of L-selectin expression was (31.86 ± 1.39) in model group, obviously higher than that of in the normal group (1.38 ±0.18) ,the CFA group( 1.50 ±0.27) and the LMS group(7.25 ±0.59) (all P <0.05) ;The inflammation cells were found around vessels and demyelination were seen in white matter of brain and spinal cords.Conclusion The expression of L-selectin should play an important role in EAE.

OBJECTIVE To observe the effects of RNA interference mediated PIK3CA gene silencing on the proliferation and invasiveness of laryngeal squamous cell carcinoma(LSCC) cells, and investigate the feasibility of PIK3CA gene as a potential therapeutic target in the treatment of LSCC. METHODS The lentiviral vector system expressing short hairpin RNA targeting PIK3CA gene(PIK3C-shRNA)was constructed and transfected subsequently into Hep-2 cells mediated by liposome in vitro. The expression of PIK3CA gene was detected by real-time RT-PCR and Western blot respectively. The proliferation of Hep-2 cells was measured by MTT, colony formation, and cell growth curve. The invasive power was determined by Boyden chamber model in vitro. RESULTS The lentiviral vector system expressing short hairpin PIK3CA- shRNA was constructed successfully. Compared with the control groups, the mRNA and protein expression of PIK3CA were significantly down-regulated(75% and 70% respectively)in the experimental group (P