Objective To investigate the effect of high concentration hydrogen gas on neurons in the rat hippocampus CA1 region during global cerebral ischemic/reperfusion injury (GCIR) Methods Four-vessel occlusion was used to establish rat model with GCIR injury. One hundred and five healthy male Sprague-Dawley rats were randomly divided into sham operation group(SH group, n = 15), model group(4-VO group, n = 45) and treatment group(4-VO+H2 group,n = 45). After 72 h and 9 d reperfusion, hippocampal CA1 region pyramidal neurons in every group were detected with Nissle staining , immunohistochemical neuron-specific nuclear protein (NeuN), specific protein antibody microglial cells (Iba1) staining and the relationship of position between neurons and microglia was observed through fluorescence double staining. We used Morris water maze to test the space orientation ability and the learning and memory ability in rats after 9 d reperfusion. Results Compared with those of 4-VO group,the neurons of hippocampus CA1 region were closer to normal in 72 h and 9 d in 4-VO+H2 group and neuron form and the number of neuron survival were increased significantly (P < 0.05);immunohistochemical staining showed that the number of neuron survival in 4-VO+H 2 group was obviously higher than that in 4-VO group (P < 0.05) and the number of microglia in 4-VO group was obviously higher than that in 4-VO+H2 group (P < 0.05). Water maze experiment showed that the swimming time in quadrant Ⅳ in 4-VO+H2 group was longer than that in 4-VO group (P < 0.05). Conclusion Inhalation of high concentration hydrogen gas has prominent protective effect on neurons of rat hippocampal CA1 region during reperfusion. The mechanism may be related with inhibiting the microglia excitation and activation during GCIR.

Objective To understand the prevalence and cause of eye diseases in the middle and old aged population in community was of important significance for the prevention and treatment of relevant diseases.The present study attempts to survey the prevalence of common eye diseases and conditions of blindness and low vision in 50 years and older population in community of Beijing.MethodsThis was an ophthalmologic epidemiology survey.The 2 833 target population was included and registered in Western Chang'an street community in door-to-door and questionnaire manner between October,2006-October,2007.Multiple factors,such as demography,general conditions,lifestyles,education degree,economical status,medical status and the awareness of relevant ophthalmic knowledge of participants were recorded and evaluated.Relevant eye examinations including uncorrected visual acuity,pinhole visual acuity,non-contact intraocular pressure test,slit-lamp,funduscopy and optometry examinations were performed in Beijing No.2 Hospital by trained ophthalmologists.Blindness and low vision were evaluated based on WHO criteria.The causes of leading-blindness and low vision were analyzed.Oral informed consent was obtained from all of the subjects.Results2 410 individuals of 2 833≥50-year-old participants received complete surveys according to the designed process in this study with an examined rate of 85.07%.The incidence of blindness and low vision in examinees was 0.79% and 5.64% respectively.The positive factors related to blindness and low vision appeared to be age,education and an awareness of relevant ophthalmic knowledge (χ~2=26.62,18.28,21.32,P＜0.001).The primary causes of blindness in examined population,in order,were cataract,glaucoma and retinopathy,and those of low vision were cataracts,refractive error and diabetic retinopathy.ConclusionIt is emphasized that early prevention and treatment for cataract,glaucoma,diabetic retinopathy and refractive errors can reduce the prevalence of blindness and low vision in people over 50 years in the Western Chang'an street community of Beijing.

OBJECTIVE:To provide reference for strengthening drug regulation in drug administration department. METH-ODS:Drug regulatory statistical yearbooks during 2011-2015 were collected. Literature analysis,content analysis,comparative analysis and secondary analysis were conducted to analyze and explore the drug production license,business license,advertising ap-proval,protection of TCM varieties,drug complaints,investigation and punishment of drug cases,etc. in statistical yearbook. RE-SULTS & CONCLUSIONS:The number of drug manufacturers and operating enterprises had been increasing year by year,while the retail chain stores in certified enterprises showed tendency to go beyond the single retail pharmacy. Compared with the down-ward trend of number of approved drug advertisements,the number of protected varieties of TCM decreased more obviously. The increase in number of drug complaints did not cause the number of investigated and punished drug cases at the same time,which showed a downward trend. It is suggested to further improve the quality and efficiency of drug regulatory work in China by strict drug production license approval,reforming drug advertising regulation and TCM varieties protection system,intensifying efforts to investigate and punish drug complaints and cases,and playing the supervision of public opinion role of the media and the masses.

Objective To investigate clinical efficacy and prognostic factors of hilar cholangiocarcinoma.Methods The retrospective case-control study was conducted.The clinicopathological data of 322 patients with hilar cholangiocarcinoma who were admitted to the Xiangya Hospital of Central South University between December 2005 and November 2015 were collected.Preoperative staging and classification of tumor and treatment planning were carried out according to the results of laboratory and imaging examinations.Observation indexes:(1) clinical features and results of assisted examinations;(2) treatments and results of pathological examination;(3) followup and survival;(4) prognostic factors analysis:gender,age,preoperative highest total bilirubin (TBil),preoperative carcinoembryonic antigen (CEA),preoperative CA19-9,preoperative CA242,preoperative CA125,treatment methods and TNM staging.The follow-up of outpatient examination and telephone interview was perfornmed to detect patients' survival up to November 2016.Survival curve was drawn using the Kaplan-Meier method.Survival and univariate analyses were done using the Log-rank test,and multivariate analysis was done using the Cox proportional hazard model.Results (1) Clinical features and results of assisted examinations:among the 322 patients,there were 301 patients with a chief complaint of jaundice.Of the 322 patients,the preoperative highest levels of TBil,DBil,ALT and AST in 322 patients were 3.9-785.2 μmol/L,1.6-410.2 μ mol/L,14.8-484.5 U/L and 21.4-539.8 U/L,respectively.Levels of ALP and GGT in 272 patients were 93.8-1 890.0 U/L and 2.0-1 832.8 U/L,respectively.Seventy-seven of 292 patients had an elevated CEA level,272 of 298 patients had an elevated CA19-9 level,153 of 260 patients had an elevated CA242 level and 86 of 260 patients had an elevated CA125 level.According to Bismuth-Corlette type,24 patients were detected in type Ⅰ,115 in type Ⅱ,55 in type Ⅲa,63 in type Ⅲb and 65 in type Ⅳ.(2) Treatments and results of pathological examination:Of the 322 patients,104 patients underwent radical resection,including 79 with hilar bile duct resection (9 combined with vascular resection and reconstruction) and 25 with extended hepatic lobectomy (16 combined with caudate lobectomy),and 218 patients underwent palliative treatments,including 134 with external biliary drainage and 84 with internal biliary drainage.Five patients were dead in the perioperative period,of which 2 died of acute liver failure,1 died of systemic infection and multiple organ failure,1 died of acute renal failure and 1 died of acute suppurative cholangitis,septic shock and disseminated intravascular coagulation.Of 263 patients receiving pathological examination,adenocarcinoma was detected in 253 patients (12 with high-differentiated adenocarcinoma,85 with moderate-differentiated adenocarcinoma,33 with low-differentiated adenocarcinoma and 123 with indefinite differentiation),mucinous adenocarcinoma in 5 patients,cholangiocarcinoma in 3 patients and neuroendocrine carcinoma in 2 patients.TNM staging of 322 patients:stage Ⅰ was detected in 8 patients,stage Ⅱ in 53 patients,stage Ⅲ in 132 patients,stage Ⅳ in 96 patients and indefinite stage in 33 patients.(3) Follow up and survival:among the 322 patients,296 were followed up for 12-132 months,with a median follow-up time of 65 months,including 94 with radical resection and 202 with palliative treatments.Among the 296 patients,the median survival time and 1-,3-,5-year survival rates were 10 months,47.1％,20.2％ and 9.5％,respectively.0f296 patients with follow-up,median survival time and 1-,3-,5-year survival rates were 31 months,84.0％,46.2％,25.0％ in 94 patients receiving radical resection and 7 months,29.9％,8.1％ and 2.3％ in 202 patients receiving palliative treatment,respectively,with a statistically significant difference between the 2 groups (x2=78.777,P＜ 0.05).Among the 94 patients receiving follow-up and radical resection,the median survival time and 1-,3-,5-year survival rates were 31 months,82.1％,45.1％,25.7％ in 73 patients undergoing hilar bile duct resection and 35 months,90.5％,49.8％,22.1％ in 21 patients undergoing hepatic lobectomy,respectively,with no statistically significant difference (x2=0.186,P＞0.05).Among the 73 patients undergoing hilar bile duct resection,median survival time and 1-,3-,5-year survival rates were 16 months,57.1％,0,0 in 7 patients combined with vascular resection and reconstruction and 34 months,84.6％,49.5％,27.5％ in 66 patients undergoing simplex hilar bile duct resection,respectively,showing a statistically significant difference (x2 =11.977,P＜ 0.05).(4) Prognostic factors analysis:results of univariate analysis showed that preoperative highest TBil,preoperative CEA,preoperative CA242,preoperative CA125,treatment methods and TNM staging were related factors affecting prognosis of patients with hilar cholangiocarcinoma (x2=25.009,18.671,9.359,33.628,94.729,77.136,P＜0.05).Multivariate analysis showed that preoperative highest TBil ≥ 342.0 μmol/L,preoperative CEA ≥ 5.00 μg/L,palliative treatments,TNM stage Ⅲ and Ⅳ were the independent risk factors affecting the poor prognosis of patients with hilar cholangiocarcinoma (HR =2.270,2.147,3.166,2.351,95％ confidence interval:1.587-3.247,1.446-3.188,2.117-4.734,1.489-3.712,P＜0.05).Conclusions Prognosis of hilar cholangiocarcinoma is still unsatisfactory.The R0 resection is the key in radical surgery.Preoperative highest TBil≥342.0 μmol/L,preoperative CEA ≥ 5.00 μg/L,palliative treatments,TNM stage Ⅲ-Ⅳ are independent risk factors affecting the poor prognosis of patients with hilar cholangiocarcinoma.

Objective To explore the protective effect of in-taking high concentration hydrogen gas on neurons and dendritic spines in hippocampus CA1 region of rats after globe cerebral ischemia/reperfusion (I/R) injury and its mechanism.Methods Four-vessel occlusion (4VO) was used to establish the models of global cerebral I/R injury in rats.One hundred and twenty healthy male Sprague-Dawley rats were randomly divided into 3 groups using a random number table:sham-operated group (inhaled 67％ N2 and 67％ O2,n=40),model group (inhaled 67％ N2 and 67％ O2 during reperfusion,n=40),and treatment group (inhaled 67％ H2 and 67％ O2 during reperfusion,n=40).After 72 h,5 and 9 d reperfusion,neuron-specific nuclear (NeuN) protein expression in the pyramidal neurons of the hippocampal CA1 region was detected with immumohistochemical staining and the positive cells were counted.And the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum were tested with colorimetry.Water maze test was used to measure the spatial orientation and memory function and Golgi staining to detect the number of dendritic spines in neurons 9 d after reperfusion.Results (1) Immunohistochemical staining of NeuN results showed that as compared with those in the model group,the neurons ofhippocampus CA1 region were significantly closer to normal with relatively intact structure,and the number of positive neurons was significantly increased in the treatment group 72 h,5 d,and 9 d after reperfusion (P＜0.05).With the reperfusion time being prolonged,the number of NeuN stained positive neurons at different time points of reperfusion in model group was gradually decreased (P＜0.05),and the numeric of the NeuN stained positive neurons at different time points of reperfusion in treatment group was slightly declined without significant difference (P＞0.05).(2) The serum SOD activity in the treatment group was significantly higher than that in the model group and sham-operated group (P＜0.05),while the MDA content in the treatment group was significantly lower than that in model group 72 h,5 d,and 9 d after reperfusion (P＜0.05).And with the reperfusion time being prolonged,the SOD activity at different time points of reperfusion showed no significant difference among the difference groups (P＞0.05).But the MDA content at different time points ofreperfusion between model group and treatment group was significantly different (P＜0.05);with the reperfusion time being prolonged,the MDA content was gradually decreased in both groups.(3) Nine d after reperfusion,water maze test found that the incubation period of treatment group was significantly shorter than that of model group (P＜0.05);the IV quadrant swimming time of space exploration in the treatment group was significantly longer than that in the model group (P＜0.05).(4) Golgi staining showed that the complexity of the neuronal dendrites branch and the number of dendritic spines of neurons in the hippocampal CA1 region of treatment group were increased than those in the model group;high-power oil microscopy indicated that the density of dendritic spines in the treatment group was significantly higher than that in the model group (P＜0.05).Conclusion In-taking of high concentrations hydrogen gas during reperfusion can definitely protect neurons in hippocampal CA1 region after globe cerebral I/R injury,and improve learning and memory function,whose mechanism may be related to hydrogen protecting the structure and function of neurons and dendritic spines,and inhibiting oxidative stress to reduce oxidative damage.

Objective: To compare the effects of different purification processes on the content of salidroside in the extract of Herba Rhodiola to provide reference for the large-scale production. Methods: Ethanol precipitation, clarifier precipitation and AB-8 macro-porous resin separation was respectively used to purify the extract of Herba Rhodiolae, and HPLC was used to determine the content of salidroside to screen the optimum purification process. Results: After the purification by AB-8 macroporous resin, the content of sali-droside was as high as 0. 272 5 mg·ml-1followed by that by ethanol precipitation (70% alcohol), and the clarifier precipitation lost more salidroside. Conclusion: The purification of salidroside by AB-8 macroporous resin separation is with high yield, good selectivi-ty, promising reproducibility and so on, which is suitable for the large-scale production of salidroside.

Objective　To establish Huashan diagnostic criteria and clinical classification system for Hirayama disease.Methods　Retrospective analysis 359 cases of puberty onset,upper extremity muscle atrophy as main clinical manifestations,and complete clinical data from September 2007 to August 2018.There were 348 males and 11 females(31.6:1 male and female)in this group.The average age of onset was 16.7±2.2 years,the average age of visits was 19.2±4.5 years,and the average duration of treatment was 29.3±45.4 months.Descriptive study of the clinical manifestations,radiologic and neurophysiological findings of this group of patients,the Huashan clinical diagnostic criteria of Hirayama disease were established by including 100% of the clinical manifestations,imaging and neurophysiological findings.According to the following parameters,the clinical classification system of Hirayama disease was proposed.The parameters specifically included:the muscle atrophy involves the upper limbs,whether the quadriplegia was active or hyperactive,the Babinski sign positive and other pyramidal tract damage,whether it was accompanied by sensory dysfunction such as upper limb numbness,muscle atrophy location,and the progress of clinical symptoms or electrophysiological examination within 6 months.Thirty patients were randomly selected from the above 359 cases.Four orthopedic surgeons who were not involved in the classification system completed the clinical classification within the specified time.The Kappa value was used for the credibility evaluation.Results　The Huashan diagnostic criteria of Hirayama disease included clinical manifestations,imaging examinations and neurophysiological examinations.The main diagnostic indicators were:1)occult onset puberty,more common in men;2)localized muscle atrophy and weakness in the upper extremities;3)compared with the cervical neutral MRI,the MRI of cervical flexion showed that spinal cord was significantly shift forward and the anterior spinal cord was narrowed or disappeared.4)MRI T2 weighting of the cervical flexion showed cyst-wall separation behind the spinal cord;5)Neurophysiological examination showed that the affected muscles were neurogenic damage.6)The affected parts are limited to the middle and lower neck segment.At the same time,it was necessary to combine imaging and neurophysiological manifestations to distinguish cervical spondylosis with upper limb muscle atrophy and motor neuron disease.According to the clinical characteristics of different patients,Hirayama disease can be divided into type I-III.Type I:72.1%,one-sided upper limb or one upper limb-based hand inner muscle and forearm muscle atrophy.According to whether progress of symptoms or electrophysiological examination was seen in the past 6 months,type I can be divided into:Ia.stable period.Regular follow-up assessment was recommended.If the disease progressed,to wear a cervical collar was suggested;surgery could be done if necessary;Ib.progression period,it was recommended to use a cervical collar,and regularly evaluate,if patients could not wear cervical collar for long,it was recommended to operate.Type II:14.2%,unilateral upper limb or one upper limb-based hand inner muscle and forearm muscle atrophy with pyramidal tract injury.Surgery was recommended.Type II:13.7%,atypical Hirayama disease,including upper limb proximal muscle atrophy,symmetrical double upper limb muscle atrophy,and sensory disturbances associated with upper limb numbness.Wear a cervical collar,and follow-up and assess closely,and choose surgical treatment if necessary.The credibility evaluation showed that the average Kappa value of the classification was 0.732(0.688-0.834),which is a basic credibility.Conclusion　The Huashan diagnostic criteria of Hirayama disease was conducive to the early diagnosis.The clinical classification system of Hirayama disease has good credibility and good clinical intervention guidance value.

In vitro multi-enzyme molecular machines that follow the designed multi-enzyme pathways, require the rational optimization and adaptation of several purified or partially purified enzyme components, in order to convert certain substrates into target compounds in vitro in an efficient manner. This type of molecular machine is component-based and modularized, so that its design, assembly, and regulation processes are highly flexible. Recently, the advantages of in vitro multi-enzyme molecular machines on the precise control of reaction process and the enhancement of product yield have suggested their great application potential in biomanufacturing. Studies on in vitro multi-enzyme molecular machines have become an important branch of synthetic biology, and are gaining increasing attentions. This article systematically reviews the enzyme component-/module-based construction strategy of in vitro multi-enzyme molecular machines, as well as the research progress on the improvement of compatibility among enzyme components/modules. The current challenges and future prospects of in vitro multi-enzyme molecular machines are also discussed.
Biotechnology ; Enzymes ; chemistry ; metabolism ; Multienzyme Complexes ; chemistry ; metabolism ; Synthetic Biology

Objective: To explore the changes of protein expression and activity of calcium/calmodulin-dependent protein kinase-II (CaMK-II) in myocardium nucleus and sarcoplasmic reticulum in experimental rabbits with heart failure (HF).
Methods: A total of 16 rabbits were divided into 2 groups: Sham group and HF group, the HF model was established by volume overload plus pressure overload.n=8 in each group and all animals were treated for 7 weeks. Left ventricular structure, hemodynamic parameters and protein expression and activity of CaMK-II in myocardium nucleus and sarcoplasmic reticulum were examined and compared between 2 groups.
Results: Compared with Sham group, HF group presented increased left ventricular mass index (LVMI) (1.32 ± 0.06) g/kg vs (3.61 ± 0.09) g/kg, LVEDP (-1.50 ± 0.50) mmHg vs (23.00 ± 2.37) mmHg, allP<0.05; while decreased left ventricular shorten fraction (37.83 ± 3.58) % vs (17.38 ± 3.13) % and LVEF (71.92 ± 4.56) % vs (38.50 ± 6.07) %, allP<0.05. The protein expression and activity of CaMK-II were both higher in HF group than Sham group, allP<0.05.
Conclusion: Increased protein expression and activity of CaMK-II in myocardium nucleus and sarcoplasmic reticulum might be one of the mechanisms for HF occurrence in experimental rabbits.

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Drug Synergism ; Escherichia coli ; genetics ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; Umbilical Cord ; cytology