Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

The paper reviewed clinical reports on the treatment of hypermenorrhea, menostaxis, and uterine bleeding with Ancong Decoction and modified Ancong Decoction. Although there were few reports concerned with clinical usage, Ancong Decoction has sound therapeutic effects, with more than 90% effective rate.

Objective To study the treatment effect on H22 tumor-bearing mice using ultrasound (US) combined with hematoporphyrin-loaded PLGA ultrasound contrast agents (HP-PLGA). Methods Hematoporphyrin-loaded PLGA ultrasound contrast agents were prepared using the technique of double emulsion, and the properties of the contrast agents were detected. Then 30 H22 tumor-bearing mice were averagely divided into 5 groups for different treatment. The growth curve of H22 tumor was depicted within 15 days after treatment. The inhibition rate of tumor quality on the 15th day and the index about apoptosis and proliferation on tumor of each group were measured, respectively. Results Self-made hematoporphyrin-loaded PLGA ultrasound contrast agents had mean size of 602.3 nm with encapsulation efficiency of 63.50% and drug loading of 2.15%. After treatment, tumors of the E group (US+HP-PLGA) grew most slowly compared with other groups. The inhibition rate of tumor quality and the apoptotic index in E group were significantly higher than those in other four groups (P＜0.05), while the proliferation index in E group was significantly lower than that in other groups (P＜0.05). Conclusion Ultrasound combined with hematoporphyrin-loaded PLGA ultrasound contrast agents could inhibit growth and promote the apoptosis of H22 tumor in mice, providing a novel strategy for sonodynamic therapy on the tumor.

Objective To optimize the formulation of a new kind of ultrasound contrast agents carrying the sensitizer of hematoporphyrin(HP)with[Poly(lactic-co-glycolic acid),PLGA]for material.Methods The technique of double emulsion was applied to produce HP loaded PLGA ultrasound microbubbles,which was optimized through orthogonal test using encapsulation efficiency for the detected index.Then morphology and distribution of HP-PLGA microbubbles were observed through light microscope and scaning electron microscope.The size,Zeta potential and the properties of releasing behavior and ultrasound imaging in vitro of Hp-PLGA contrast agents were detected.Results The optimization parameters were picked out as 10 mg/ml for concentration of HP,40 mg for PLGA,and 1/5 for volume ratio of water inside to dichloromethane.The optimized HP-PLGA contrast agents were spheric with the mean size of 602.3 nm,and Zeta potentiaI of-(17.1±1.6)mV.The drug loading and encapsulation efficiency of HP-PLGA were(2.15±0.15)%and(63.5±2.6)%,respectively.And the releasing behavior of HP-PLGA contrast agents in vitro was that after an obvious release of about 35.1%in former 24 h,there were 86.5%HP-PLGA released within 14 days.The ultrasound imaging of HP-PLGA could be enhanced obviously in vitro.Conclusions The self-made HP-PLGA ultrasound microbubble might be a useful tool for delivering sensitizer and thus provide a novel strategy for sonodynamic therapy on tumor.

To observe the clinical effect of Yisui decoction plus western medicine in treating multiple system atrophy patients, totally 65 patients from China-Japan Friendship hospital during 2008-2012 with complete clinical data and received consecutive traditional Chinese medicine and western medicine treatment for more than 3 months were observed changes of traditional Chinese medicine symptom score, part 1 of unified multiple system atrophy rating scale, orthostatic hypotension before treatment and after 3 months treatment. After 3 months treatment, total effective rate of traditional Chinese medicine symptom was 70.8%. Compared with before treatment, score of part 1 of unified multiple system atrophy rating scale was obviously reduced after 3 month treatment (P < 0.001). Ex- cept swallow function without significant improvement, the remaining projects of unified multiple system atrophy rating scale were im- proved obviously (P < 0.05), of which the most obvious differences were orthostatic symptoms, falls and intestinal function (P < 0.001). Orthostatic hypotension after 1 month treatment and 3 month treatment was obviously better than before treatment (P < 0.001). There was no significant difference in orthostatic hypotension between 1 month treatment and 3 month treatment. The research results show that Yisui decoction plus western medicine has a certain effect on improving clinical symptoms of multiple system atrophy patients, especially has a significant effect on orthostatic hypotension, and can maintain a stable clinical effect in a certain period of time.
Adult ; Aged ; Humans ; Hypotension, Orthostatic ; drug therapy ; Male ; Medicine, Chinese Traditional ; adverse effects ; methods ; Middle Aged ; Multiple System Atrophy ; drug therapy ; Retrospective Studies ; Treatment Outcome

Objective To investigate the CT signs of small intracranial aneurysm and the application value of multi-slice spiral CT angiography (MSCTA) before and after clipping surgery and in follow-up.Methods MSCTA imaging data of 81 patients with highly suspected small intracranial aneurysms from March 2013 to March 2015 were analyzed retrospectively.The confirmed patients underwent clipping surgery of small intracranial aneurysms.The imaging findings before and after clipping surgery were analyzed and compared with those by DSA and what observed during surgery.Results The relative accuracy,sensitivity and specificity of MSCTA over DSA and surgical diagnosis were 93.83％,93.75％ and 94.12％respectively,and MSCTA gained high relative consistency over DSA and surgical diagnosis (Kappa=0.825).The accuracy of MSCTA for diagnosing small intracranial aneurysm was not significantly different from those by DSA and surgical diagnosis.MSCT plain scan mainly showed subarachnoid or intracranial hemorrhage,and found 71 rhomboic or saccular aneurysms in 60 patients before operation.Totally 64 patients went through clipping surgery,60 ones had satisfactory results in the reexamination by MSCTA 2 weeks after surgery,and there were no abnormality found in 36 patients in 6 to 12-month follow-up.Conclusion MSCTA can provide abundant information for clipping surgery of small intracranial aneurysms,and can be used as the preferred imaging method for postoperative evaluation and follow-up.

ObjectiveTo investigate the application of dexmedetomidine attenuating hemodynamic fluctuation in patients with cerebral aneurysm during anesthesia induction.Methods Sixty patients undergoing elective interventional procedure with cerebral aneurysm were divided into low dose of fentanyl group (group LF),high dose of fentanyl group (group HF),and low dose of fentanyl and dexmedetomidine combination group(group FD) by random digits table method with 20 cases each.Before induction of anesthesia,the patients in group FD received dexmedetomidine 1 μ g/kg for 10 minutes,and the others received 0.9％ sodium chloride with the same volume.During anesthesia induction,fentanyl 3 μ g/kg in group LF and group FD,and fentanyl 5 μ g/kg in group HF.Other anesthetics were equalized.Systolic blood pressure (SBP),diastolic blood pressure (DBP) and heart rate (HR) were monitored and recorded at 3 min utes after lying on table (T0),before intubation (T1),immediately and 3 minutes after intubation (T2,T3).The differences between the maximum and the minimum of SBP,DBP and HR were calculated in these time points as fluctuation values,named as △SBP,△DBP and △HR.Results△SBP,△DBP and △HR in group FD [ (26.9 ± 14.8) mm Hg( 1 mm Hg =0.133 kPa),(10.7 ± 8.9) mm Hg,(12.5 ± 4.3 ) times/min ]were lower than those in group LF [ (40.4 ± 15.6) mm Hg,(20.3 ± 9.4) mm Hg,(30.1 ± 15.0) times/min ](P ＜ 0.05 ),as well as △SBP and △HR in group HF [ (29.8 ± 16.8 ) mm Hg,( 19.5 ± 7.4) times/min ]were lower than those in group LF(P ＜ 0.05).While △HR in group FD was lower than that in group HF (P ＜0.05).The usage of atropin in three group had no statistical significance during anesthesia induction (P =0.364),but more ephedrine was used in group HF than in group LF [30％(6/20) vs.5％(1/20),P=0.032 ].Conclusion Dexmedetomidine 1μ g/kg injected before anesthesia induction,which could prevent intubation reaction,blood pressure serious falling after intubation,and provide more stable hemodynamics,is particularly applicable for anesthesia induction in patients with cerebral aneurysm.

OBJECTIVETo explore the repair of Xiangsha Liujunzi Decoction (XSLJZD) on interstitial cells of Cajal (ICC) and gap junction (GJ) in the gastric muscular layer of rats of Pi-qi deficiency syn- drome (PQDS).

METHODSPQDS was established using purgative method with bitter and cold drugs in 30 healthy Wistar rats. After successful modeling they were randomly divided into the treatment group and the model group, 15 in each group. Another 15 healthy Wistar rats were recruited as the healthy control group. Rats in the treatment group were gastric administered with XSLJZD at 2 mL/100 g body weight, once daily for 14 successive days. Equal volume of normal saline was gastrically administered to those in the healthy control group and the model group. The gastric muscle tissues were taken out before modeling, before intervention, and after intervention, respectively. Ultrastructural changes of ICC and GJ were observed using transmission electron microscope (TEM). The number and distribution of Connexin43 (Cx43) were detected using immunohistochemistry.

RESULTSResults of TEM indicated that compared with the healthy control group, both ICC and GJ in the model group showed obvious injury. ICC and GJ were apparently repaired after intervention in the treatment group. Compared with the same group before modeling, the integrated optical density (IOD) of the Cx43 expression significantly decreased in the model group before and after intervention (P <0.05). Compared with before intervention, the IOD of the Cx43 expression significantly increased in the treatment group (P <0.05). Compared with the healthy control group, the IOD of the Cx43 expression significantly decreased in the model group before and after intervention (P <0.05). Compared with the model group, the IOD of the Cx43 expression significantly increased in the treatment group (P <0.05).

CONCLUSIONSUltrastructures of ICC and GJ in the gastric muscular layer of rats of PQDS were obviously damaged. XSLJZD could repair the structural damage of ICC and GJ in the gastric muscle tissues of rats of PQDS.

Animals ; Connexin 43 ; Drugs, Chinese Herbal ; pharmacology ; Gap Junctions ; Interstitial Cells of Cajal ; drug effects ; Leydig Cells ; Male ; Muscle, Smooth ; Qi ; Rats, Wistar ; Syndrome

BackgroundDry eye is a multi-factorial-induced tear film and ocular surface disorder.Immunoinflammation plays a key role in the pathogenesis of dry eye.As inhibitor of the cyclo-oxygenase pathway,nonsteroidal anti-inflammatory drugs play an anti-inflammatory and anti-hypersensitivity role,and it can be a potential treatment for dry eyes.ObjectiveThis study was to investigate the effectiveness of nonsteroidal anti-inflammatory drugs (0.1％topical pranoprofen) on moderate to severe dry eyes and its mechanism.MethodsThis was a small sample of randomized controlled clinical trial.Thirty right eyes of 30 patients with moderate to severe dry eyes were included in the study according to the diagnosis criteria and randomized into two groups.The patients of the trial group received topical administration of 0.1％ pranoprofen plus 0.1％ sodium hyaluronate,and those of the control group received the topical 0.1％ sodium hyaluronate only.Ocular surface inflammation index scores (OSDI) and ocular surface fluorescine staining (OSS) scores were measured under the slit lamp,and tear film break-up time (BUT),Schirmer Ⅰ test values were evaluated.The expression of human leucocyte antigen-DR (HLA-DR) and CD11b in conjunctiva epithelial cells were detected by impression cytology and flow cytometry (FCM).All the indexes were compared between the two groups before and after treatment.Informed consent was obtained from all patients.ResultsThere were no significant differences in terms of age and gender and their baseline values between the trial group and control group (t=0.412,P=0.684;x2=0.240,P=0.624),and so were all the indexes (P＞0.05).Compared with the control group,the OSDI,OSS scores and cells positive for HLA-DR were lowered but the BUT was delayed in the trial group on day 15 ( t=2.43,P=0.03;t=2.83,P=0.01;t=3.29,P=0.00;t=3.23,P=0.00 ).No significant differences were found in the Schirmer Ⅰ test value and CD11b expression between these two groups (t=0.17,P=0.87;t=0.28,P=0.79).The OSDI,OSS scores and BUT were significantly improved,and the number of cells positive for HLA-DR were reduced 15 days after administration of drugs in comparison with before treatment in the trial group ( t =12.30,10.70,6.10,7.92,P =0.00 ).However,there were no comparable alteration seen in these indexes before and after the usage of drugs in the control group ( P＞0.05).Positive correlations were found in HLADR expression with OSDI and OSS ( r =0.601,P =0.018 ; r =0.586,P =0.022 ) and a negative correlation in HLADR expression with BUT (r=-0.697,P=0.004) on day 15 in the trial group.ConclusionsTopical usage of 0.1％ pranoprofen is beneficial for remitting the ocular signs and symptoms in moderate to severe dry eyes.This study illustrates that topical usage of 0.1％ pranoprofen can down-regulate the expression of inflammatory markers in conjunctival epithelial cells.

Objective To investigate public network attention to air pollution and their desire to prevent haze. Methods Daily Air Quality Index(AQI) and PM_2.5 concentration data and daily Baidu Index data were obtained in 31 provinces of China from January 1^st, 2015 to December 31^st, 2017.The Baidu indexes of air pollution and its protection were used to compose public′s network attention index for analysis on its spatial distribution in the 31 provinces and for linear regression research on air pollution, its protection and its network attention impact factors. Results With Baidu search index, it was noted that the network attention to air pollution was significantly higher than to air pollution protection.And a significant difference was found in the public network attention to air pollution between different provinces.It was found that education and consumption level of residents significantly affected public network attention to air pollution, while AQI and consumption level of residents significantly affected public network attention to air pollution hazards protection behaviors. Conclusions Even though the public has already paid high attention to air pollution, the awareness of air pollution hazards protection behaviors is still weak.Well-designed education campaigns are needed to enhance public awareness of air pollution and motivate the public to take actions to reduce pollution air exposure, especially in those provinces with a high AQI and PM_2.5concentration.