The patch clamp recording technique in vivo is a blind patch clamp recording methods to record the current of the spinal or cereral neurons of anaesthesia ( or awake) animals. This technique can be used to study the synaptic function and plasticity in central nervous system in vivo in order to understand the physiological properties of the ion channels from an integrated point of view. The advantage of this technique has already presented itself in the study of the synaptic transmission and nervous network. Nowadays, in vivo patch whole-cell recording technique in combination with other techniques is becoming a common method in the research fields.

BACKGROUNDEndothelial progenitor cells (EPCs) transplantation is a promising therapeutic strategy for ischemic retinopathy. The current study aimed to establish a simple, reliable and fluorescent labeling method for tracking EPCs with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) in laser-injured mouse retina.

METHODSEPCs were isolated from human umbilical cord blood mononuclear cells, cultivated, and labeled with various concentrations of CFSE. Based on fluorescence intensity and cell morphology, a 15 minutes incubation with 5 µmol/L CFSE at 37°C was selected as the optimal labeling condition. The survival capability and the apoptosis rate of CFSE-labeled EPCs were measured by Trypan blue staining and Annexin V/PI staining assay respectively. Fluorescence microscopy was used to observe the label stability during the extended culture period. Labeled EPCs were transplanted into the vitreous cavity of pigmented mice injured by retinal laser photocoagulation. Evans Blue angiography and flat mounted retinas were examined to track the labeled cells.

RESULTSEPCs labeled with 5 µmol/L CFSE presented an intense green fluorescence and maintained normal morphology, with no significant changes in the survival capability or apoptosis rate after being labeled for 2 days, 1 and 4 weeks. The fluorescence intensity gradually decreased in the cells at the end of 4 weeks. Evans Blue angiography of the retina displayed the retinal capillarity network clearly and fluorescence leakage was observed around photocoagulated spots in the laser-injured mouse model. One week after transplantation of labeled EPCs, the fluorescent cells were identified around the photocoagulated lesions. Four weeks after transplantation, fluorescent tube-like structures were observed in the retinal vascular networks.

CONCLUSIONEPCs could be labeled by CFSE in vitro and monitored in vivo for at least 4 weeks, and participate in the repair of injured retinal vessels.

Animals ; Cells, Cultured ; Endothelial Cells ; chemistry ; cytology ; Fluoresceins ; chemistry ; Fluorescent Dyes ; chemistry ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Microscopy, Fluorescence ; Retina ; cytology ; Stem Cells ; chemistry ; cytology ; Succinimides ; chemistry

OBJECTIVETo observe the intervention and mechanism of Weinaokang capsule (WNK) on cognitive impairment rats induced by chronic cerebral hypoperfusion.

METHODThe bilateral common carotid arteries of rats were ligated. Rats were intragastrically administrated with WNK 4 weeks after the ligation. After 8 weeks of administration, persistent time of rats on finding the platform in MORRIS water maze was measured, meanwhile, content of acetylcholine (ACh) in brain tissue was measured by HPLC-ECD, activity of acetylcholine esterase (AChE) and superoxide dismutase (SOD), content of malonaldehyde (MDA) was measured by colorimetric method.

RESULTAfter 4-8 weeks of administration, the time of finding the platform in WNK group was significantly shorter than in the model group, activity of AChE in brain tissue was significantly reduced (P < 0. 05), content of ACh was significantly increased (P < 0.05-0.01), activity of SOD was significantly reinforced (P < 0.05).

CONCLUSIONWNK capsule can improve the cognitive impairment induced by chronic cerebral hypoperfusion, and its mechanism may be associated with reinforcing the capability of scavenging free radical and protecting the cholinergic system.

Acetylcholine ; metabolism ; Acetylcholinesterase ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; Cerebrovascular Disorders ; drug therapy ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Maze Learning ; drug effects ; Memory ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo observe the effects of maternal exposure to nano-alumina during pregnancy on the neurodevelopment in offspring mice.

METHODSFemale ICR mice began to be exposed to nano-alumina 10 d before mating, and the nano-alumina exposure lasted till offspring mice were born. All the female mice were randomly divided into 5 groups: solvent control group (saline), nano-carbon group (11.76 mg/ml), micro-alumina group (50 mg/ml), 50 nm alumina group (50 mg/ml), and 13 nm alumina group (50 mg/ml). All the mice were treated by nasal drip (10 µl/time) 3 times daily till offspring mice were born. Physiological indices, reflex and sensory function test, endurance test, Morris water maze test, positioning and navigation test, and open field test were used to evaluate the neurodevelopment of newborn mice.

RESULTSOn day 28, the body weight of 13 nm alumina group (16.73±4.04 g) was significantly lower than that of solvent control group (20.45±2.50 g) (P<0.01); the 13 nm alumina group had significantly delayed time to ear opening compared with the solvent control group (4.91±0.78 d vs 4.45±0.50 d, P<0.01); compared with the solvent control group, the nano-carbon group, micro-alumina group, 50 nm alumina group, and 13 nm alumina group had significantly delayed time to eruption of teeth (10.05±0.23 d vs 10.32±0.48 d, 10.75±0.45 d, 10.32±0.47 d, and 10.79±0.49 d, P<0.05 or P<0.01). On days 4 and 7 after birth, compared with the solvent control group, other groups had significantly decreased proportions of mice which passed the cliff avoidance test (P < 0.05 or P < 0.01). On days 12 and 14 after birth, compared with the solvent control group, the nano-carbon group, 50 nm alumina group, and 13 nm alumina group had significantly reduced pre-suspension time in the endurance test (P < 0.05 or P < 0.01). The Morris water maze and positioning and navigation tests showed that the 13 nm alumina group had a significantly increased 5 d incubation period compared with the solvent control group (P < 0.05); compared with the solvent control group, other groups had significantly reduced numbers of platform crossings (P < 0.05 or P < 0.01). The open field test showed that the nano-carbon group and 13 nm alumina group had reduced numbers of rearings compared with the solvent control group (P < 0.05); compared with the solvent control group, other groups had significantly reduced numbers of modifications (P < 0.01).

CONCLUSIONMaternal exposure to nano-alumina (13 nm) during pregnancy has inhibitory effects on the physical development and early behavioral development in newborn mice and can also inhibit the learning and memory abilities and adaptability to new environment in offspring mice. The neurodevelopmental toxicity of nano-alumina to newborn mice increases as the particle sizes of nano-alumina decrease, which has been demonstrated by the endurance test and number of rearings.

Aluminum Oxide ; toxicity ; Animals ; Animals, Newborn ; Behavior, Animal ; Body Weight ; Female ; Maternal Exposure ; Maze Learning ; Mice ; Mice, Inbred ICR ; Motor Activity ; Nanostructures ; toxicity ; Pregnancy

OBJECTIVETo evaluate the clinical efficacy of intradermal injection of methylene blue for treatment of moderate to severe acute thoracic herpes zoster and prevention of postherpetica neuralgia in elderly patients.

METHODSSixty-four elderly patients with herpes zoster were randomized to receive a 10-day course of intradermal injection of methylene blue and lidocaine plus oral valaciclovir (group A, 32 cases) and intradermal injection of lidocaine plus oral valaciclovir (group B).Herpes evaluation index, pain rating index, incidence of postherpetic neuralgia, and comprehensive therapeutic effect were compared between the two groups at 11, 30 and 60 days after the treatment.

RESULTSThe baseline characteristics were comparable between the two groups (all P>0.05). Compared with that in group B, the time for no new blister formation, blister incrustation and decrustation, and pain relief was significantly shortened in group A (P<0.05) with also obviously lower pain intensity after the treatment. The incidence of postherpetic neuralgia was significantly lower in group A than in group B at 30 days (P<0.05), but not at 60 and 90 days after the treatment. The total clinical response rate was 93.8% in group A, much higher than that in group B (62.5%, P<0.05).

CONCLUSIONIntradermal injection of methylene blue can effectively shorten the disease course, reduce the pain intensity and prevent the development of postherpetic neuralgia in elderly patients with herpes zoster.

Acyclovir ; administration & dosage ; analogs & derivatives ; therapeutic use ; Aged ; Herpes Zoster ; complications ; Humans ; Incidence ; Injections, Intradermal ; Lidocaine ; administration & dosage ; therapeutic use ; Methylene Blue ; administration & dosage ; therapeutic use ; Neuralgia, Postherpetic ; therapy ; Pain Measurement ; Valine ; administration & dosage ; analogs & derivatives ; therapeutic use

OBJECTIVETo explore the effects of silencing HERC4 on the proliferation, apoptosis, and migration of cervical cancer cell line Hela and the possible molecular mechanisms.

METHODSThree HERC4-specific small interfering RNAs (siRNAs) were transfected into Hela cells, and HERC4 expression in the cells was examined with Western blotting. CCK-8 assay, annexin V-FITC/PI assay, and wound healing assay were used to assess the effect of HERC4 silencing on the proliferation, apoptosis and migration ability of Hela cells. The expression levels of cyclin D1 and Bcl-2 in the cells were detected using Western blotting.

RESULTSTransfection of siRNA-3 resulted in significantly decreased HERC4 protein expression (P<0.01). HERC4 silencing by siRNA-3 markedly suppressed the proliferation and migration of Hela cells, increased the apoptosis rate (P<0.01) and reduced the expression levels of cyclin D1 and Bcl-2 (P<0.01).

CONCLUSIONSilencing of HERC4 efficiently inhibits the proliferation, migration, and invasion of Hela cells in vitro, and the underlying mechanisms may involve the down-regulation of cyclin D1 and Bcl-2.

Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclin D1 ; metabolism ; Down-Regulation ; Female ; HeLa Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Ubiquitin-Protein Ligases ; genetics ; metabolism ; Uterine Cervical Neoplasms ; pathology

OBJECTIVETo investigate the relationship between occupational stress and depression in migrant workers.

METHODSMigrant workers in the textile industry were selected as subjects, and the self-made Occupational Stress Questionnaire and Self-rating Depression Scale were used to investigate the sex, age, seniority, educational level, and marital status of these subjects. Data analysis was performed by independent-samples t test, analysis of variance, Spearman rank correlation analysis, and stepwise multiple regression analysis.

RESULTSSex, seniority, and educational level were not influential factors for depression scores. The lower age group had a higher moderate depression score than the higher age group; the unmarried group had a higher moderate depression score than the married group. Severe depression was negatively correlated with decision-making power, psychological job demands, social support, and external pay-return, but positively correlated with skills and internal input; moderate depression was positively correlated with psychological job demands and external pay, but negatively correlated with other factors; mild depression was negatively correlated with all factors. The stepwise regression analysis showed that the influential factors for depression included, from major to minor, supervisor support, skills, internal input, and colleague support, according to the standardized regression coefficients; internal input was the contribution factor, and the remaining ones were negative factors.

CONCLUSIONAmong migrant workers, certain mental health problems exist, and occupational stress is associated with depression.

Adolescent ; Adult ; Aged ; Depression ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; epidemiology ; Stress, Psychological ; epidemiology ; Surveys and Questionnaires ; Transients and Migrants ; Young Adult

OBJECTIVETo establish a rabbit model of silicotic pulmonary fibrosis and to investigate the effect of cordyceps sinensis in this model.

METHODSThirty healthy male white rabbits were randomly divided into control group, silicosis model group, and intervention group. The rabbits in silicosis model group and intervention group received endotracheal perfusion of silicon dioxide suspension (120 mg/kg), and the control group was treated with the same volume of saline. All the rabbits were sacrificed 30 days later. The lung coefficient was calculated by comparing the lung weight and body weight; the right lung tissue was stained with hematoxylin-eosin (HE). The content of hydroxyproline in lung tissue was measured by alkaline hydrolysis. The mRNA levels of transforming growth factor beta 1 (TGF-β₁) and mothers against decapentaplegic homolog 7 (Smad7) in rabbit lung sections were determined by real-time PCR.

RESULTSNo abnormalities were observed by HE staining in the lung tissues of control group, while fibrosis and silicotic nodules were discovered in the silicosis model group and intervention group. The lung coefficient and the content of hydroxyproline in lung tissue were significantly higher in the silicosis model group than in the control group and intervention group (P < 0.05 or P < 0.01). Compared with the control group, the silicosis model group and intervention group had significantly increased TGF-β₁ mRNA levels but significantly reduced Smad7 mRNA levels (P < 0.02). Compared with the silicosis model group, the intervention group had a significantly reduced TGF-β₁ mRNA level but a significantly increased Smad7 mRNA level (P < 0.05).

CONCLUSIONCordyceps sinensis is able to reduce the expression of TGF-β₁ mRNA and increase the expression of Smad7 mRNA in lung tissues of rabbits with silicotic pulmonary fibrosis, and thus postpone the progression of fibrosis.

Animals ; Cordyceps ; chemistry ; Disease Models, Animal ; Lung ; metabolism ; Male ; Pulmonary Fibrosis ; drug therapy ; metabolism ; Rabbits ; Silicosis ; drug therapy ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo compare the therapeutic effects of the greenlight photoselective vaporization of prostate (PVP) and transurethral electrovaporization resection of prostate (TUVP) for the treatment of symptomatic benign prostatic hyperplasia (BPH).

METHODSOne hundred and sixty-three cases of BPH were treated with PVP and TUVP. All patients were followed up with International Prostatic Symptom Score (IPSS), quality of life (QOL), blood loss, operative time, indwelling catheterization, mean Qmax, residual urinary volume (RUV) and operative complications.

RESULTSIPSS, QOL, Qmax and RUV were significantly improved after either of the procedures (P < 0.05), no significant difference in the improvement of subjective symptoms and objective signs had been noted with the different procedure (P > 0.05). Mean operative time was (37 +/- 15) min for TUVP and (45 +/- 28) min for PVP, the resection time was longer for PVP than TUVP (P > 0.05), but the intraoperative bleeding and catheterization time were less for PVP than TUVP (P < 0.05). Postoperative complications were less for PVP than TUVP (P < 0.05). The incidence of hematuria in TUVP group had been 41.4%, and urinary irritation after PVP group was 55.2% (P < 0.05).

CONCLUSIONSPVP has the same therapeutic effect as TUVP and less adverse side effects than TUVP. It is a new technique for the treatment symptomatic BPH.

Aged ; Aged, 80 and over ; Follow-Up Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Prostate ; pathology ; surgery ; Prostatic Hyperplasia ; surgery ; Quality of Life ; Transurethral Resection of Prostate ; methods ; Treatment Outcome

Objective To analyze the general characteristics of voluntary blood donors, and to provide the basis for establishing effective recruitment mechanism. Methods The information of all whole blood donors (including mobile blood collection cars on the streets , blood donation houses and team blood donors) from 2006 to 2015 was obtained from the Zhejiang blood management information system. The sex, age, occupation, weight, and the amount of blood donation at a time of blood donors were descriptively analyzed. Results There were 5299700 frequencies of donating blood in Zhejiang Province from 2006 to 2015. The annual frequency was on the rise, with an average annual growth rate of 2.99％. There were 3098500 frequencies of male and 2201300 of female with the ratio of 1.4 :1. The frequency of female has declined since 2010. The majority of voluntary blood donors ranged from 18 to 45 years old, and the percentage was 91.44％ . The ratios of staff, students and workers were 14.61％, 12.27％, 10.09％, respectively. The annual frequency of staff was on the rise, with an average annual growth rate of 7.71％. The frequency of workers was on the decline, and the frequency of students fluctuated over the years. The ratio of blood donation 400 mL at a time was 41.23％. For 400 mL blood donation at a time, the ratio of male donors (48.86％) was significantly higher than the female donors (30.50％, P<0.001) . In addition, the ratio of blood donation 400 mL at a time was proportional to age and body weight (P<0.001) . Conclusion In Zhejiang Province, the frequency of blood donation has increased year by year. Staff, students and workers with 18 to 45 years old are the main force of blood donation. The frequency of female blood donation has a tendency of dropping, and the ratio of blood donation 400 mL at a time is relatively low. It's necessary to strengthen the propaganda of blood donation.