Objective To observe the effect of transcranial low frequency electrical stimulation on the contents of monoamines in ischemic area of rats with middle cerebral artery occlusion(MCAO).MethodsPermanent MCAO model of Wistar rat was established with silk thread enveloped with polyammoniacum.The ischemic areas received various intensity of transcranial low frequency electrical stimulation for 1 hour in rats underwent 1 hour of ischemia.Then the affected tissue was processed with fluorospectrophotometry to determine the contents of dopamine(DA),noradrenalin(NE) and 5-hydroxytryptamine(5-HT).ResultsCompared with the sham-operation group,the contents of DA,NE and 5-HT in ischemic area of MCAO model rats decreased obviously(all P<0.01),while all three monoamines investigated in the sham-operation group with transcranial low frequency electrical stimulation had no significant change.In the MCAO groups stimulating with lower(10 V) and middle(30 V) intensity transcranial low frequency electrical field,the contents of DA,NE and 5-HT in ischemic area had no significant increase.But in the MCAO group stimulating with high(50 V) intensity transcranial low frequency electrical field,the contents of DA,NE and 5-HT in ischemic area increased significantly(P<0.05).ConclusionSome degree of intensity transcranial low frequency electrical field stimulation can increase the contents of DA,NE and 5-HT in ischemic area of rats subjected to MCAO.

Objective:To analyze the levels of individual doses to radiation workers receiving medical radiation exposure in hospitals in Jiangxi province, so as to provide reference for radiation protection.Methods:The radiation workers of different types in medical institutions in Jiangxi were investigated from 2014 to 2018, involving diagnostic radiology, radiotherapy, nuclear medicine and interventional radiology. Statistical analyses were made of the individual doses to medical radiation workers in the province.Results:The total number of monitored workers were 23 833 with average annual individual effective dose of 0.316 mSv. NR1 and NR5, the ratios of the number of workers receiving annual individual doses exceeding 1 and 5 mSv to the total monitored workers were 4.32% and 0.10%, respectively. During the five-year period, the average annual individual effective dose showed a upward and then downward trend, with peak appearing in 2017. A total of 17 909 diagnostic radiologists were monitored, accounting for 75.14% of the total number of monitored workers. The average annual individual effective dose received by workers was 0.329 mSv, the highest, in interventional radiology, followed by diagnostic radiology 0.318 mSv, nuclear medicine 0.283 mSv, and radiotherapy 0.269 mSv, respectively. There were statistically significant differences among radiation workers of different types ( χ2=489.39, P<0.001). Conclusions:The average annual individual effective dose to the radiation workers in medical institutions in Jiangxi province meets the relevant national standards, but with an upward trend in nuclear medicine. The higher levels of individual doses to radiation workers are seen in primary and secondary hospitals. It should be strengthen the oversight of radiation workers in nuclear medicine and in primary and secondary hospitals.

Objective To investigate the expression of nucleotide-binding oligomerization domain protein 2 (NOD2)in serum of patients with hepatocellular carcinoma (HCC),and to analyse the roles of NOD2 in HCC development and its clinical diagnostic value.Methods This study including 66 patients with HCC in the hospi-tal from March 1,2013 to December 31,2014 and 61 healthy controls.Serum NOD2 levels were determined by enzyme-linked immunosorbent assay (ELISA).Analysis of significance was performed with rank sum test using SPSS statistical 16.0 software.Results Serum levels of NOD2 in HCC patients were 171 pg/ml,significantly higher than that of healthy controls(95 pg/ml,Z =-5.00,P =0.00),and the serum NOD2 levels were correla-ted with clinical stage of HCC (H=56.26,P =0.00).Compared with the serum NOD2 levels in stageⅠ,Ⅱpatients (106 pg/ml)and healthy controls (95 pg/ml),the serum NOD2 level in stage Ⅲ and Ⅳ (220 pg/ml) were significantly increased (χ2 =31.24,P =0.00;χ2 =47.23,P =0.00),but the expression of NOD2 in stageⅠandⅡwere nearly equal to that of the healthy controls (χ2 =0.36,P =0.83).The ROC analysis revealed that the best diagnostic cutoff-point of serum NOD2 levels for predicting the Ⅲ and Ⅳ stages of HCC was 148.78 pg/ml,meanwhile corresponding sensitivity was 89.1% and specificity was 77.0%.Additionally,corre-lation analysis demonstrated that there was no significant correlation between NOD2 and alpha-fetal protein (r =0.44,P =0.14).Survival curves obtained that the survival time of HCC patients with NOD2 serum concentrations≥ 200 pg/ml was significantly less than that <200 pg/ml (χ2 =15.32,P <0.05).Conclusion NOD2 is highly expressed in the serum of HCC patients,especially in advanced patients,which is possibly involved in the development of HCC and has the potential to become an effective marker used for HCC diagnosis.

Objective:To study the enhancing effect of bFGF-targeted antisense phosphorothioate oligonucleotide (APO)on the chemosensitivity of human laryngeal squamous carcinoma cell line Hep2 to Doxorubicin,5-Fluorouracil, and Cisplatin.Methods:bFGF-specific APO was designed,constructed and transfected into Hep2 cells with jetPEI (polyethyleneimine).Expression of bFGF mRNA was evaluated by semi-quantitative RT-PCR after transfection;immuno- cytochemical method was used to examine the expression of bEGF expression before and after transfection of Hep2;the in- duction of cell apoptosis was analyzed by flow cytometry;cell proliferation was then analYzed by MTT assay after treatment with bFGF-specific APO or chemotherapeutic drugs,or a combination of both.Results:bFGF-specific APO inhibited the growth of Hep2 cells in a dose-and time-dependent manner,with the peak inhibitory rate being 25.5%.The expression of bFGF mRNA and protein decreased by 52.0% and 41.1%,respectively.The apoptosis rate of Hep2 cells was 20.5% after transfection,bFGF-specifie APO reduced the 50% inhibitory concentration of Doxorubicin,5-Fluorouracil,and Cis- platin in Hep2 cells by 75.5%,83.5% and 65.4%,respectively.Conclusion:bFGF-specific APO can enhance the chemosensitivity of Hep2 cells,which paves a new way for potential biologic chemotherapy of laryngeal squamous carcino- ma.

Objective To explore the clinical outcome of improved technique of Gamma nail in the treatment of intertrochanteric hip fracture of the elderly patients. Methods From March 2002 to October 2006.39 patients with intertrochanteric hip fracture were operated by improved technique of Gam-ma nail.There were 18 males and 21 females at average age of75.7 years(67_98 years).There were 6 patients with type A1 fracture,24 with type A2 fracture and 9 with type A3 fracture according to AO/ASIF classification.Of all.36 patients(92.3%)had osteoporosis.The operation improvements included the following points:(1)The patients were placed at the lateral decubitus position with the fractured limb on the uppermost,with flexion of knee and hip of 60°.The normal hip and knee were flexed as possible.(2)One-off indirect traction reduction was used after general anesthesia. no requirement of continuous mechanical traction.(3)C-arm image intensifier was employed to obtain normal and lateral projections.Results Of all,35 patients were followed up for a mean period of3 years and 2 months, ranging from 6 months to 5 years and 2 months.Operation data showed incision length of(4.3±1.2)cm,mean opera-tion time of(46±10)minutes,intraoperative bleeding volume of(65±26)ml and intraoperative X-ray exposure of(3.0±2.1)times.Postoperative recovery data showed survival in one-year follow up,with ambulation time of(10.5±3.6)days and fracture union time of(10.9±2.1)weeks.Mean Parker's score wag(6.9±3.2)points 6 months after operation. Conclusions Improved technique of Gamma nail can shorten operation duration,reduce operative trauma and bleeding,reduce X-ray exposure and im-prove success rate of surgery.as facilitates early pest-operative recovery and reduces the perioperative mortality rate of the elderly.

Objective To observe the change of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression during 3T3-L1 adipocyte differentiation as well as other reference gene expressions.Methods The mRNA expressions of several common reference genes were detected by real time-PCR on day 0,1,3,5,and 7 of 3T3-L1 adipocyte differentiation.Western blot was used to confirm the protein expressions of three common reference genes.Results (1) GAPDH and transferrin receptor(TFRC) mRNA expressions were significantly increased during adipocyte differentiation.GAPDH mRNA level was increased by 5.7,7.6,22.0,and 24.5 folds on day 1,3,5,and 7 after induction of adipocyte differentiation,but no apparent changes of β-actin,α-tubulin,peptidylprolyl isomerase A (PIPA),and 18S mRNA expressions were detected.The expression changes of key transcript factors for adipocyte differentiation such as PPARγ2,C/EBPα,and C/EBPβ were under-estimated by real time-PCR if GAPDH was chosen as the reference gene.Western blotting results showed that the GAPDH protein level increased gradually during adipocyte differentiation,especially on day 5 and 7 after adipocyte differentiation.There were no obvious changes of β-actin and α-tubulin protein expressions.(2) Berberine significantly inhibited mRNA and protein expressions of GAPDH in the process of adipocyte differentiation.GAPDH mRNA levels were reduced by 68.1％ and 66.3％ on day 5 and 7 after induction of adipocyte differentiation,but with no significant change in other reference genes.Conclusion It is not suitable for GAPDH to be used as an endogenous reference gene during 3T3-L1 adipocyte differentiation.

OBJECTIVETo investigate the effects of exogenous TGF-β3 on the expression of endogenous TGF-b3 in hepatic stellate cell (HSC).

METHODSHSCs were cultured and divided into two groups: TGF-β3 group and blank control group, the cells of TGF-β3 group were exposed to TGF-b3 (10 ng/ml), whereas the blank control group was not treated. The cells were incubated in the presence of exogenous TGF-β3 and then (1) were harvested at 0h, 1h, 2h, 4h, 12h, 24h, and real time PCR was performed to detect the mRNA expression of endogenous TGF-β3. (2) The cells were collected at 0h, 1h, 6h, 12h, and western-blot was used to detect the protein synthesis of endogenous TGF-β3 in HSC; (3) The cell culture supernatant was harvested at 0h, 1h, 2h, 4h, 8h, 14h, 24h, and ELISA was performed to measure the total protein of extracellular TGF-β3; HSCs were treated with TGF-β3 (10 ng/ml) for 2h. The cells were then incubated in serum-free medium and the cell culture supernatant was harvested at 2.25h, 2.5h, 3h, 4h, 6h, 10h and 14h. ELISA was used to detect the extracellular secret ion of endogenous TGF-β3 by HSCs.

RESULTS(1) Exogenous TGF-β3 treatment induced a marked increase in TGF-β3 mRNA expression. By 2h of exogenous TGF-β3 treatment, maximal TGF-β3 mRNA expression levels (2.796 ± 0.518) of 2.74 fold above control values (1.022 ± 0.038) was reached (P < 0.05). Thereafter, TGF-β3 mRNA expression level declined, and the expression level was maintained at level of 1.45-fold for at least 10h and was 1.18-fold above control values by 24h TGF-β3 treatment (P < 0.05); (2) No significant difference about the intracellular protein expression level of endogenous TGF-β3 was found between two groups. (P > 0.05); (3) The total expression level of TGF-β3 reached a peak [(18.931 ± 2.904) ng/ml] at 4h after TGF-β3 treatment (1.89-fold higher than basic TGF-β3 (10 ng/ml). After that, it slowly declined. The expression peak [(0.835 ± 0.027) ng/ml] induction of extracellular secreted TGF-β3 was at 3h (32.12-fold higher than control [(0.026 ± 0.022) ng/ml], (P < 0.05). Thereafter, TGF-β3 slowly decreased after the peak time, and their expressions were still statistically significant as compared to the control (P < 0.05).

CONCLUSIONExogenous TGF-β3 could increase the expression of endogenous TGF-β3 mRNA and extracellular secreted TGF-β3 protein obviously.

OBJECTIVETo investigate the effects of propofol combined with indomethacin on the contractile function of isolated human pulmonary arteries.

METHODSHuman pulmonary artery preparations were obtained from patients undergoing surgery for lung carcinoma. The intrapulmonary arteries were dissected and cut into rings under microscope for treatment with propofol or propofol combined with indomethacin. In each group, the rings were divided into endothelium-intact and endothelium-denuded groups and mounted in a Multi Myograph system. In propofol group, the rings were preconstricted by U46619 to induce a sustained contraction, and propofol (10-300 mmol/L) was then applied cumulatively. In the combined treatment group, the rings were pretreated with indomethacin (100 µmol/L) for 30 min before application of U46619 to induce sustained contraction, and propofol (10-300 µmol/L) was added cumulatively after the tension became stable.

RESULTSPropofol (10-100 µmol/L) induced constrictions at low concentrations and caused relaxations at higher concentrations (100-300 µmol/L) in the pulmonary artery rings with prior U46619-induced contraction. Propofol caused stronger constrictions in endothelium-intact rings [EC=4.525∓0.37, Emax=(30.44∓2.92)%] than in endothelium-denuded rings [EC=4.699∓0.12, Emax=(31.19∓5.10)%, P<0.05]. Pretreatment of the rings with indomethacin abolished constrictions, and the relaxation was more obvious in endothelium-intact group [pD=3.713∓0.11, Emax=(98.72∓0.34)%] than in endothelium- denuded group [pD=3.54∓0.03, Emax=(94.56∓0.53)%, P<0.05].

CONCLUSIONPropofol induces constriction at low concentrations and relaxation at high concentrations in human intrapulmonary arteries with U46619-induced contraction. Indomethacin abolishes the constriction induced by propofol in isolated intrapulmonary arteries, suggesting that propofol potentiates U46619-mediated pulmonary vasoconstriction by promoting the concomitant production of prostaglandin by cyclooxygenase in pulmonary artery smooth muscle cells, and the mechanism for its relaxation effect may partly depend on the endothelium.

In order to produce recombinant human anti-HBsAg Fab antibody in Pichia pastoris, the recombinant yeast was fermented using fed-batch system in a 30 L bioreactor. The fermentation temperature was 30 degrees C, the pH was 5.0 approximately 5.3, and the DO was 20% approximately 30%. The recombinant Fab antibody was purified from crude culture supernatant by ion exchange and analyzed by SDS-PAGE and western blot and ELISA. When the absorbance (OD600) of broth reach 300 at the end of fed-batch phase, the induced phase was initiated. The results showed that recombinant human anti-HBsAg Fab antibody was high-level expressed in recombinant Pichia pastoris using a fed-batch fermentation system. Both chains of the Fab were successfully expressed upon methanol induction. After 192 h of induction, the expression level of recombinant Fab (soluble) reached 412 mg/L. The recombinant Fab antibody was purified effectively by ion-exchange chromatography from the fermentation supernatant to a purity of 95%. And the affinity activities of the purified recombinant Fab antibdy and fermentation supernatant were detected, and both of them showed high affinity activities. The results demonstrated that recombinant human anti-HBsAg Fab antibody could be high level produced by fed-batch fermentations in Pichia pastoris. Which can be efficiently used in industrial production.
Fermentation ; Hepatitis B Antibodies ; biosynthesis ; isolation & purification ; Hepatitis B Surface Antigens ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; isolation & purification ; Pichia ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification

Postmortem chemistry is becoming more and more essential in routine forensic pathology and has made considerable progress over the past years. Biochemical analyses of vitreous humor, blood, urine and cerebrospinal fluid may provide important information in determining the cause of death or in elucidating forensic issues. Postmortem chemistry may be essential for the determination of cause of death when morphological methods (diabetes mellitus, alcoholic ketoacidosis and electrolytic disorders) cannot detect the pathophysiological changes involved in the death process. It can also provide many information in other forensic situations, including myocardial ischemia, sepsis, inflammation, infection, anaphylaxis and hormonal disturbances. The most recent relevant research advances on glucose metabolism, liver function, cardiac function, renal function, sepsis, inflammation, infection, anaphylaxis and hormonal aspect are hereby reviewed.
Anaphylaxis ; Autopsy/trends* ; Biomarkers/analysis* ; Body Fluids/chemistry* ; Death ; Diabetes Mellitus ; Forensic Pathology/methods* ; Humans ; Postmortem Changes ; Sepsis ; Vitreous Body