OBJECTIVE:To provide new identification method for processed medicinal material Chinese polyphaga(Eupolyph-aga sinensis,Steleophaga plancyi) and their adulterants by establishing molecular identification method based on Cytb genes. METHODS:The total DNA of Chinese polyphaga and their adulterants was extracted using modified saturation sodium chloride method. The Cytb genes of all samples were amplified with PCR using general primers REVCB2H and REVCBJ. The phylogenetic tree of all samples was constructed with Neighbor-Joining(NJ)method using MEGA 5.1 software. The sequences of the Cytb gene of all sampled were compared by using DNAMAN sofetware. The difference between genuine product and their adulterants were analyzed,and the specific primers Esin-F and Esin-R were designed for molecular identification in different regions. RESULTS:DNA extracted from processed medicinal insects was successful to amplify Cytb gene segments. The phylogenetic tree of all sam-ples was consistent with their genetic relationship. A fragment was amplified only from genuine product but not from other adulter-ants with the designed specific primers Esin-F and Esin-R. CONCLUSIONS:DNA extraction method from processed Chinese polyphaga and their adulterants have been established. Designed specific primers are highly specific to genuine product Chinese polyphaga,and can be used for the identification of Chinese polyphaga and their adulterants.

Autoimmune Diseases ; chemically induced ; Female ; Humans ; Interferons ; adverse effects ; Purpura, Thrombocytopenic, Idiopathic ; chemically induced ; Young Adult

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.

Objective To elucidate the MRI appearance of prostatic abscess,the DWI and enhanced MRI features.Methods 12 cases of prostatic abscesses were retrospectively analyzed,the clinical symptom mainly manifested as lower urinary tract symptoms and fever.All of the patients were given routine MR examination including DWI sequence,6 patients received further enhanced MR examination.Results In the 12 cases,there were 4 cases behaved as single type,8 cases as multifocal type.The abscess showed iso-or slightly hypo-signal intensity on T1 WI,hyper-signal intensity on T2 WI,markedly high signal intensity on DWI and correspond-ing markedly low signal intensity on ADC.Complete abscess walls showed iso-or slightly hyper signal on T1 WI,hypo-signal inten-sity on T2 WI.The mature abscess walls were thin and smooth,which showed homogeneously ring enhanced in 4 cases.The imma-ture abscess walls showed uneven thickness and moderately enhanced in 2 cases.Septum in the abscess could be found in 4 cases, which showed similar enhancement to the abscess walls,while the abscess cavity showed non-enhanced.Abscesses involved the sur-rounding structures in 2 cases,the involved area showed obvious hyper-signal on T2 WI fat-suppression sequence.Conclusion DWI is the best sequence in the diagnosis of prostatic abscess,the markedly high signal intensity on DWI is the characteristic sign.The enhanced MRI showed the walls and septa clearly,the extent and involvement of adjacent structures.The multi-parametric MRI is a prominent procedure in the diagnosis of prostatic abcess.

Aim To investigate the interactions be-tween the neuroinflammation caused by lipopolysaccha-ride(LPS) and brain CYP2E1.Methods The human cholinergic neuroblastoma cell line IMR-32 was treated with LPS ( 0.1 mg · L-1 , 1.0 mg · L-1 ) , and the LDH and SOD activities were determined after 24 h in-cubation .In order to determine the roles of MAPK sig-naling pathway in the regulation of CYP 2E1 by LPS, the IMR-32 cells were treated with p38 pathway inhibi-tor SB203580 or ERK pathway inhibitor U 0126 for 45 min before the incubation with LPS .The human do-paminergic neuroblastoma cell line SH-SY5Y with CYP2 E1 over-expression was established . The LDH and SOD activities were determined in SH-SY5 Y cells over-expressed CYP2 E1 and control cells treated with LPS(0.1 mg· L-1 , 1.0 mg· L-1 ) for 24 h.Results
The levels of LDH in IMR-32 cells treated with high-dose LPS were increased by 1.38-fold ( P <0.01 ) compared with the control group , and the levels of SOD reduced by 15.0%( P <0.01 ) .Compared with the control, CYP2E1 mRNA and protein levels in IMR-32 cells treated with high-dose LPS were increased by 1.25-fold(P<0.01) and 1.19-fold(P<0.05).The up-regulation of CYP2E1 by LPS could be attenuated by SB203580 and U0126 pretreatment.Compared with the control cells, the CYP2E1 over-expression in-creased LDH levels by 1.28-fold ( P<0.01 ) and de-creased SOD levels by 3.53-fold ( P<0.01 ) after the low-dose of LPS treatment .The CYP2E1 over-expres-sion increased LDH levels by 1.54-fold ( P <0.01 ) and decreased SOD levels by 2.17-fold( P<0.01) af-ter the high-dose of LPS treatment , compared with the control cells.Conclusions LPS can induce CYP2E1 mRNA and protein levels , and the p38 and ERK sig-naling pathway may be involved in the regulation .The elevated CYP2 E1 levels aggravate the damage to neuro-nal cells caused by LPS .Brain CYP2E1 may be an im-portant contributing factor to the pathological process of neuroinflammatory injury .

Objective To study the effect of allicin on the growth and radiosensitivity of human pancreatic carcinoma BXPC3 cells.Methods BXPC3 cells were exposed to X-rays in the presence or absence of allicin.Cell proliferation was measured by MTT assay.Cell cycle distribution and apoptosis were detected by flow cytometry assay.Cell radiosensitivity and the influence of allicin on it was evaluated by colony formation assay.The expressions of Bax and Bcl-2 proteins were assayed by RT-PCR and Western blot.Results IC50 values of allicin on cell growth were 76.24,58.34 and 43.58 μmol/L under 12,24 and 48 h treatment,respectively.Treatment of cells with allicin obviously inhibited cell growth after irradiation and hence increased radiosensitivity (t =2.74,P ＜ 0.05).This treament also enhanced radiation-induced cell cycle arrest at G2/M phase (t =11.41,P ＜0.05),apoptosis induction (t =12.36,P ＜ 0.05),and Bax expression (t =4.83,P ＜ 0.05),but it decreased Bcl-2 expression (t =3.69,P ＜ 0.05).Conclusions Allicin could inhibit cell growth,induce cell cycle arrest and apoptosis via Bax/Bcl-2 pathway and hence increases radiosensitivity of BXPC3 cells.

Objective To investigate the influence factors of central venous catheter related infections (CVC-RI) in postoperative patients with gastric carcinoma treated with parenteral nutrition,in order to provide scientific basis for prevention and treatment.Methods Using the retrospective study method,150 patients who treated with parenteral nutrition after operation of gastric carcinoma from December 2013 to March 2014 were selected.Information including age,sex,anamnesis,catheter indwelling time,infusion rate of nutrient solution,and so on were collected.Patients with central venous catheter related infections were set as infection group,patients without infection as the control group,and the group logistic regression analysis was used to analyze the collected data.Results 21 patients in infection group had CVC-RI,and the incidence was 14.00％ (21/150),while 129 patients in control group had CVC-RI.Univariate analysis showed there was significant difference between infection group and control group in age,anamnesis,catheter indwelling time and infusion rate of nutrient solution (x2=17.126,5.447,12.795,7.973,P＜0.05),however there was no significant difference in sex,health insurance,the success rate of catheterization and whether glutamine was in nutrient solution (P＞0.05);the group logistic regression analysis showed that age (OR=15.927,95％CI 3.305-76.756,P＜0.05)and catheter indwelling time (OR=3.511,95％CI 1.028-11.992,P＜0.05) were independent risk factors for CVC-RI,however infusion rate of nutrient solution (OR=0.140,95％CI 0.033-0.597,P＜0.05) was a protective factor.Conclusions For the postoperative patients with gastric carcinoma treated with parenteral nutrition,CVC-RI is closely related to age,catheter indwelling time and infusion rate of nutrient solution.So reducing catheter indwelling time,restoring the enteral nutrition under condition permission as so on as possible can reduce infection of elderly patients.

Objective To compare the effect of 4 different cold and hot properties of traditional Chinese medicine (TCM) on body temperature and related factors of yeast induced fever rats,and discuss the thermoregulatory mechanism of cold and hot properties of TCM.Methods 108 male SD rats were randomly divided into a normal group,a yeast-induced group,a R.palmatum treated group,a C.chinensis treated group,a Euodia ruticarpa treated group,and a Alpinia officinarum Hance treated group,with 18 rats in each group.Pyrexia model was induced by injecting yeast suspension subcutaneously on rat.At the 4h,8h and 12h after injection of yeast,the rats were sacrificed,and the blood and hypothalamus were collected.The levels of prostaglandin E2 (PGE2),cyclic adenosine 3',5'-monophosphate (cAMP) and arginine vasopressin (AVP) in hypothalamus and plasma were detected by ELISA assay.Results At the 4h after injection of yeast,the temperature of rats in the model group began to rise,and it reached the peak at 8h,while RheumpalmatumL and Coptis chinensis could significantly reduce the body temperature of yeast-induced rat (P＜ 0.01 or P＜ 0.05).At 8h,the levels of PGE2 and cAMP in hypothalamus increased significantly [respectively (31.55 ± 9.88) pg/mg and (0.17±0.03) pmol/mg] compared with the normal group,while the level of AVP (0.14±0.02) pmol/ml in plasma reduced (P＜0.05).Compared with model group,at 8h RheumpalmatumL and Coptis chinensis could significantly lowered PGE2 [respectively (113.65± 18.60) pg/mg and (127.72 ± 15.75) pg/mg,P＜ 0.01 or P＜0.05],and cAMP [respectively (0.69±0.08) pmol/mg and (0.74±0.10) pmol/mg,P＜0.05] in hypothalamus,and increased AVP levels [respectively (1.08 ± 0.12) pmol/ml and (0.91 ±0.01) pmol/ml,P＜0.05 or P＜0.01] in plasma.Euodia ruticarpa and Alpinia officinarum had no significant effect on both body temperature and the levels of inflammatory factors.Conclusion The two cold property traditional Chinese medicines,R.palmatum and C.chinensis,could significantly reduced the body temperature of yeast-induced rats,which may be related to its effective regulation on levels of PGE2 and cAMP in hypothalamus and AVP in plasma,however,the two hot property traditional Chinese medicine,Euodia ruticarpa and Alpinia officinarum Hance,had no related effects.

Objective:To establish and evaluate a newly established method of enzyme-linked immu-nosorbent assay (ELISA) for measuring human autoantibody to folate receptor (FR).Methods: Folate receptor was extracted and purified from healthy woman placenta tissues .The protein was coated on 96-well plates.Goat monoclonal antibody was used as detecting antibody to set up the indirect ELISA proce -dure.The sensitivity, precision and linearity of the method were evaluated .Further, the method was compared with the ELISA method with commercialized bovine folate binding protein ( FBP) by determi-ning autoantibody levels in 24 individuals .Results:The measuring range of the standard curve was from 6 .25 ×10 -4 to 8 ×10 -2 ( the IgG concentration of pooled plasma from healthy donors was defined as 1 ) . The lowest detectable level was 3.13 ×10 -4 .The intra-and inter-assay coefficients of variations were 2.74%-8.07% and 4.16% -8.23%, respectively.Linearity test results were considered within acceptable limits.The data from FBP-ELISA and FR-ELISA were highly correlated ( r=0.954, P <0.001);The value from FR-ELISA was higher by 14% than that from FBP-ELISA.Conclusion: The ELISA method for measuring human autoantibody IgG to folate receptor was successfully established using human FR as coating protein .The method is sensitive and repeatable and can be used in large-scale population study .

Objective To analyze the clinical manifestations and electroencephalogram (EEG)characteristics of agyria-pachygyria for its early diagnosis,treatment and prognosis judgment in clinical practice.Methods The clinical manifestations and EEG features of twenty-four patients with agyria-pachygyria who were diagnosed by CT or magnetic resonance imaging(MRI) at Pediatric Neurology of Children's Hospital of Chongqing Medical University from July 2004 to July 2013 were retrospectively analyzed.Results Of twenty-four patients,eighteen cases were diagnosed as diffuse agyria-pachygyria and six cases were diagnosed as partial agyria-pachygyria.The clinical features were mainly manifested as mental retardation (twenty-four patients),and motor retardation (twenty-four patients),and epilepsy (eighteen patients).All of the twenty-four patients had abnormal EEG pattern which were mainly three tapes.Type Ⅰ had diffused high amplitude alpha and beta activity in all cortical regions,frontal-central,or parietal-occipital region (fourteen patients).Type Ⅱ showed alternating high amplitude bursts with sharp and slow waves (seven patients).Type Ⅲ was characterized by high amplitude spike or sharp wave activity generalized or multifocal distribution and δ,θ wave mixing graphics (twelve patients).Nine of twenty-four patients showed two or three EEG characteristic patterns in an awake-asleep EEG recording.During the follow-up of 1-8 years old,twelve of the thirteen patients who were diagnosed as epilepsy in diffuse agyria-pachygyria had refractory epilepsy,mainly with infantile spasms or Lennox-Gastaut syndrome.One of the five patients who was diagnosed as epilepsy in focal agyria-pachygyria had refractory epilepsy,mainly for partial epilepsy secondary generalized seizures.There was a significant difference between them (P =0.008).Eighteen of twenty patients who had moderate-severe mental retardation or dyskinesia were diagnosed as diffuse a gyria-pachygyria,while two were focal agyria-pachygyria.Both of them had a significant difference (P =0.005).Conclusions Agyria-pachygyria is a brain malformation caused by neuronal migration abnormality.Diffuse agyria-pachygyria is presented with serious clinical manifestations and poor outcome while the clinical manifestation of focal agyria-pachygyria is relatively mild and epilepsy could be controlled by antiepileptic drugs or epilepsy surgery.These characteristics of EEG patterns along with clinical findings could provide important evidence for early diagnosis,timely treatment and prognosis judgment of agyria-pachygyria.