From genomics to proteomics, and then to miRNA, all without exception showed that proteins are the substance which directly regulate life. Comparing with genomics, the study of proteins is more difficult for its great variety, complicated modification and complicated construction. Recently, some new techniques for protein assay and research have provided us, for example, Fluorescence labeling, SELDI, SPR and optical protein chip, of which, SELDI SPR and optical protein chip are label-free. SELDI protein chip technique and its newly developed applications are briefly reviewed here.

Objective To evaluate the efficacy of itopride hydrochloride combined with alpha-lipoic acid in treatment of diabetic gastroparesis.Methods One hundred and ten patients with diabetic gastroparesis were randomly assigned to two groups.Sixty patients received α-lipoic acid plus itopride (α-lipoic acid 0.6 g intravenously for 14 d plus itopride 50 mg orally t.i.d), and 50 cases received oral itopride only.The clinical symptoms including postprandial abdominal fullness, belching, acid reflux, anorexia, nausea and vomiting were observed;the gastric emptying rate was tested, and plasma motilin and serum ganstrin levels were measured groups, before and after two weeks of treatment.Results The clinical symptom scores of combination therapy group (4.62 ± 3.46 vs.12.26-± 3.43) and control groups (5.43-± 3.65 vs.12.31-± 3.37) were improved after treatment.The overall effective rate of combination group was 91.6％ and that of itopride therapy group was 78.0％ (x2 =4.098, P ＜ 0.05).Gastric emptying rate of combination group [(76.9 ± 9.7) ％ vs.(37.3 ± 10.2) ％] was increased more markedly than that ofitopridegroup[(57.2-± 10.4)％ vs.(36.8-± 11.2)％] (t =10.221, P ＜0.05).Plasma motilin and serum ganstrin levels in combination group [(640.2 ± 64.4) ng/L vs.(554.0 ± 67.6) ng/L, (152.6 ± 20.3)ng/L vs.(96.8 ± 22.2)ng/L] were reduced more markedly than those in itopride group [(636.3 ±66.4)ng/L vs.(589.4 48.3)ng/L, (153.5-± 19.4) ng/L vs.(120.9 ± 19.7)ng/L] respectively (t =3.100, 5.959, all P ＜ 0.05).Conclusion For patients with diabetic gastroparesis, α-lipoic acid and itopride combination therapy can significantly improve symptoms and gastric emptying rate, decrease motlin and gastrin levels;and the efficacy of combination therapy is better than that of itopride monotherapy.

Objective To evaluate the effect of sevoflurane on the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in hippocampal neurons of rats.Methods Forty-eight pathogen-free healthy adult Sprague-Dawley rats of both sexes,weighing 210-280 g,aged 3-4 months,were divided into 4 groups (n=12 each) using a random number table:control group (group C) and 1%,3% and 5% sevoflurane groups (S1,S2 and S3 groups).In S1,S2 and S3 groups,1%,3% and 5% sevoflurane were inhaled for 1 h,respectively.Morris water maze test was performed at 1 and 7 days after anesthesia to assess the cognitive function.Six rats in each group were sacrificed after the end of the test.The brains were then removed and the hippocampi were isolated for detection of the expression of α7nAChR (by Western blot) and contents of tumor necrosis factor-alpha and interleukin-1 (by enzyme-linked immunosorbent assay).Results Compared with group C,the number of crossing the platform was significantly decreased at 1 and 7 days after anesthesia in group S1,the time of staying at the target quadrant was significantly shortened,the number of crossing the platform was decreased,and the rate of time of staying at the original platform quadrant and rate of swimming distance at the original platform quadrant were decreased at 1 and 7 days after anesthesia in S2 and S3 groups,and the contents of tumor necrosis factor-alpha and interleukin-1 were significantly increased,and the expression of α7nAChR was down-regulated at 1 and 7 days after anesthesia in S1,S2 and S3 groups (P<0.05).Compared with group S1,the expression of α7nAChR was significantly down-regulated at 1 and 7 days after anesthesia in S2 and S3 groups (P<0.05).Compared with group S2,the expression of α7nAChR was significantly down-regulated at 1 day after anesthesia in group S3 (P<0.05).There was no significant difference in each parameter of Morris water maze test among group S1,group S2 and group S3 (P>0.05).Conclusion The mechanism by which sevoflurane induces inflammatory responses in hippocampi is related to down-regulation of α7nAChR expression in hippocampal neurons of rats.

Objective To review and analyze the prenatal diagnosis and treatment of fetal bronchopulmonary sequestration (BPS).Methods Data of 68 BPS fetuses in the Department of Fetal Medicine,Guangdong Women and Children Hospital and Health Institute from January 20,2009 to January 31,2013 were reviewed.All of the 68 fetuses were diagnosed by ultrasound and volume to head circumference ratio (CVR) was measured.Postnatal surgeries at neonatal or infantile period were performed and babies were followed-up.Results With antenatal ultrasonography,fetal chest solid or hyper-echoic mass was found in all 68 cases at an average of 24 weeks of gestation (18-30 weeks) and were diagnosed as BPS after at least twice ultrasound scans.CVR was measured in these 68 cases,among them 56 cases (82.4％) were ＜1.6,and 12 cases (17.6％) were ≥1.6.Among the 12 cases with CVR≥1.6,seven developed fetal hydrops.Except for two terminated pregnancies because of CVR≥ 1.6 with fetal hydrops,all of the rest 66 cases continued their pregnancies till birth and 58 (87.9％) neonates were asymptomatic and eight (12.1％) were symptomatic.The mass was undetectable with ultrasonography in 18 fetuses during the third trimester,however,the mass was identifed again under CT scan after birth in 14 cases (another four cases unexamined).There were totally 45 cases fully recovered after lobectomy or BPS resection after birth,among which five cases underwent emergency surgery in neonatal period.Pathology results showed that 12 cases were intra-lobar and 33 cases were extra-lobar type.Totally,six cases were diagnosed as BPS under diaphragm.Sixty cases were followed up at three months to two years after birth,and all developed well and CT scan indicated normal pulmonary function.Conclusions Most BPS fetuses without hydrops have a good outcome.Fetus with BPS could wait for full term delivery following professional evaluation and standard protocol.However,the prognosis of BPS with fetal hydrops depends on prenatal evaluation and early surgical intervention.And the effect of early surgical intervention in BPS neonates is satisfied.CVR is the most important index for prenatal evaluation of BPS and CT scan after birth is critical for those babies with undetectable mass by prenatal ultrasound.

Animals ; Drugs, Chinese Herbal ; pharmacology ; PC12 Cells ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; drug effects ; physiology ; Rats

OBJECTIVETo study the effect of Qinghuang Powder (QHP) combined Chinese herbs for Pi-strengthening and Shen-reinforcing (CHPSSR) on hypoxia-inducible factor 1alpha (HIF-1alpha) in bone marrow mononuclear cells of myelodysplastic syndrome (MDS) patients.

METHODSChanges of HIF-1alpha in bone marrow mononuclear cells of MDS patients were detected in 25 MDS patients treated by QHP combined CHPSSR using flow cytometry. Meanwhile, 13 healthy subjects were recruited as the control group. Changes HIF-1alpha levels in various serial bone marrow mononuclear cells were detected.

RESULTS(1) Among the 25 patients in the treatment group, there were 19 patients effective and 6 patients ineffective, with the total effective rate being 76%. (2) Compared with before treatment, levels of peripheral blood WBC, Hb, PLT, and ANC significantly increased in the treatment group after treatment, showing statistical difference (P < 0.05, P < 0.01). (3) Compared with before treatment, the HIF-1alpha mean fluorescence intensity was enhanced in bone marrow lymphocytes, monocytes, granulocytes, and nucleated red blood cells of the treatment group after treatment (P < 0.05, P < 0.01). Compared with the control group, the HIF-1alpha mean fluorescence intensity was weakened in bone marrow lymphocytes, monocytes, and nucleated red blood cells of the treatment group before treatment; while it was obviously enhanced in granulocytes (P < 0.01). But after treatment the HIF-1alpha mean fluorescence intensity increased more in the granulocytes of the treatment group than in those of the control group (P < 0.01), but there was no statistical difference in bone marrow lymphocytes, monocytes, or nucleated red blood cells.

CONCLUSIONQHP combined CHPSSR could improve HIF-1alpha levels in bone marrow lymphocytes, monocytes, granulocytes, and nucleated red blood cells of MDS patients, thus improving Hb levels of MDS patients, and improving their anemia and correlated symptoms.

Adolescent ; Adult ; Aged ; Arsenicals ; therapeutic use ; Bone Marrow ; Bone Marrow Cells ; drug effects ; metabolism ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Middle Aged ; Monocytes ; drug effects ; metabolism ; Myelodysplastic Syndromes ; drug therapy ; metabolism ; Phytotherapy ; Young Adult

Objective:To investigate the effect and mechanisms of Phloroglucinol ( PG) on renal ische-mia and reperfusion injury (IRI).Methods:Forty-eight male Wistar rats were divided into 3 groups (16 rats per group):sham operated, saline-treated I/R (I/R), and PG-treated I/R (PG).I/R model:Af-ter removing the right kidney , renal I/R injury was induced by clamping the left renal artery for 45 min followed by reperfusion.The rats were administered with PG (30 mg/kg, intraperitoneally) or saline 15 min before renal ischemia .The blood and kidneys were harvested 6 and 24 h after reperfusion .Renal function and histologic changes of serum creatinine (SCr) and blood urea nitrogen(BUN)were assessed. Malondialdehyde (MDA),catalase (CAT),superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were measured.Nuclear factor-kapa B ( NF-κB) and caspase-3 in the kidneys were also measured.Results:SCr and BUN were (103.9 ±10.4) μmol/L and (15.2 ±1.0) mmol/L in I/R group, and (81.8 ±13.4) μmol/L and (11.5 ±1.2) mmol/L in PG group 6 h after reperfusion .SCr and BUN were (154.9 ±12.1) μmol/L and (28.1 ±1.4) mmol/L in I/R group, and (103.8 ±5.9)μmol/L和(16.0 ±1.0) mmol/L in PG group 24 h after reperfusion.PG treatment significantly attenua-ted renal dysfunction and histologic damage caused by I /R injury(P<0.05).The I/R-induced elevation in kidney MDA level decreased , where as reduced kidney SOD ,CAT and GSH-Px were increased .What is more , the apoptotic tubular cells , the levels of active caspase-3 ,and active nuclear factor kappa B dra-matically decreased after PG treatment .Conclusion:PG protects murine kidney I/R injury by suppres-sing oxidative stress , inflammation , and cell apoptosis .

AIM: To construct an eukaryotic expression vector of human single-chain variable fragment ~against hepatitis B virus core protein (anti-HBc ScFv) and detect its expression in HepG2 cells. METHODS: Anti-HBc ScFv genes were amplified from the plasmids abstracted from positive clone and inserted into pEGFP-c1 vector that contained green fluorescent protein gene. The recombinant plasmids were transfected into HepG2 cells, and resistant clones were obtained by G418 selection. The expression of the gene of fusion protein was determined by fluorescent invert microscope and ELISA. RESULTS: Recombinant plasmids were successfully constructed. The plasmid transfected HepG2 cells were obtained by G418 selection. Specific fluorescence was observed in HepG2 cells 48 hours after transfection. ELISA analysis confirmed the expression of anti-HBc ScFv in the cells. CONCLUSION: The construction of human anti-HBc ScFv eukaryotic expression vector and its expression in HepG2 cells lay the foundation for advanced research of intracellular anti-HBc ScFv.

Aim To study the protective effect of CDPS on acute aging mouse model induced by D-galactose (D-gal) and its mechanism.Methods (1) The acute aging mouse model was induced by D-gal.After CDPS (25、50、100 mg·kg-1) treatment, the improving effect on learning and memory in mice was examined in vivo.(2) We also established the aging model on PC12 cells in vitro.After CDPS treatment (150、200 mg·L-1), the level of p-CREB in the nucleus was detected by Western blot, and the content of cAMP, PKA and brain derived neurotrophic factor (BDNF) levels were examined by the Elisa kits.Moreover, cAMP, PKA and BDNF were detected in PC12 cells under the condition that H89, the inhibitor of PKA, co-cultured with PC12 cells after CDPS treatment.(3) The UPLC/Q Exactive MS method was developed for determining the concentration of glutamic acid, dopamine and norepinephrine, which secreted in PC12 cells after CDPS treatment.Results (1) In vivo, CDPS significantly improved the memory impairment in aging mice induced by D-gal in the Morris assay.(2) In vitro, CDPS could significantly increase the expression of p-CREB (P<0.05), PKA, cAMP and BDNF (P<0.05).The H-89 abolished the increase of p-CREB (P<0.05), PKA, cAMP and BDNF (P<0.05) in PC12 aging cells induced by D-gal after CDPS treatment.(3) CDPS increased the release of dopamine, norepinephrine, and glutamate secreted in PC12 cells.Conclusion CDPS could significantly improve the learning and memory ability on aging mouse model in vivo, and reversed the damage in PC12 cells induced by D-gal by activating cAMP/PKA/CREB signal cascade, increase the expression of BDNF, and increasing modestly the release of excitatory neurotransmitter.

Objectives To analyze the incidence of cranial nerve injury in patients after receiving carotid endarterectomy ( CEA) in a single-center and to investigate its correlation with surgical experiences. Methods The clinical data of patients underwent CEA at Beijing Xuanwu Hospital, Capital Medical University from January. 2001 to December 2013 were analyzed retrospectively. Cranial nerve injury was assessed at day 7 after procedure according to the clinical symptoms,and they were followed up at 1,3,6, and 12 months. The incidence of permanent cranial nerve injury was evaluated. The incidences of permanent cranial nerve injury were further analyzed at two time periods ( from January 2001 to September 2011 and from October 2011 to December 2013 ) . Results ( 1 ) A total of 598 consecutive patients treated with CEA were enrolled,and 15 (2. 5%) of them had cranial nerve injury,including 2 (0. 3%) facial nerve injury,7 (1. 2%) hypoglossal nerve injury,and 6 (1. 0%) vagus nerve injury. Only 1 case (0. 2%) did not recover completely at 6 months after procedure. ( 2 ) The patients with cranial nerve injury were 10 (3. 2%,10/308) and 5 (1. 7%,5/290) respectively from January 2001 to September 2011 and from October 2011 to December 2013). There was no significant difference (P < 0. 05). Conclusion The incidence of cranial nerve injury was low after CEA,and most patients could recover completely. The increased surgical experiences did not show the reduction of cranial nerve injury rate evidently after CEA.