OBJECTIVETo study the effects of Kuntai Capsule (KTC), a Chinese herbal preparation, on the quality of life (QOL), breast distending pain and vaginal bleeding in women at early stage of menopause.

METHODSA total of 104 women at early stage of menopause, 54 had their uterus existed (Ue) and 50 in-existed (Ui), were enrolled, and they were randomized to the KTC group and the control group, with equal cases of Ue and Ui in each. The KTC group was treated with KTC 4 capsules twice a day; the control group treated with premarin 0.45 mg per day and for those of Ue 2 mg medroxyprogesterone additionally, with the remedies medicated orally for 1 year. All the testees were asked to record everyday their own condition of breast pain and vaginal bleeding and followed-up every 3 months to fulfill the Menopause Specific Quality of Life questionnaire. Ultrasonic examination on pelvis and breast as well as endocrine hormone assays of estradiol (E2) and follicle-stimulating hormone (FSH) were performed before and after the medication term.

RESULTSEffects of treatment in the two groups were different insignificantly in terms of QOL. The women were benefited in vasomotor and physical domains from the 3rd month of medication, and the psychosocial domain was also improved (for Ui initiating from the 3rd month and for Ue from the 6th month). In the domain of sexual life, KTC showed its favorable effect only on Ue beginning from the 9th month, but not on Ui; while all subjects in the control group had their sexual life improved from the 3rd month. In domain of breast pain, the occurrence at various time points between the two groups was insignificantly different, only that the severity in Ue of the control group was more significant from the 1st to 3rd month than that in the KTC group. As for the domain of vaginal bleeding, the uterine membrane was basically normal in both groups either before or after medication, but the incidence and lasting days from the 1st to 3rd month in Ue of the KTC group were significantly lower than those of the control group. Levels of E2 and FSH were not significantly changed after medication in the KTC group, while in the control group, E2 significantly increased and FSH significantly decreased in the women of Ue (P <0.05).

CONCLUSIONKTC could evidently improve the QOL of women at the early stage of menopause, and is of high safety, with less adverse reaction of breast pain and vaginal bleeding, and shows few impact on sexual hormones.

Adult ; Breast ; pathology ; physiology ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; pathology ; Humans ; Menopause ; drug effects ; psychology ; Middle Aged ; Pain ; drug therapy ; pathology ; Quality of Life ; Vaginal Diseases ; drug therapy ; pathology ; psychology

A stimulate method for determination of polybrominated diphenyl ethers ( PBDEs) and derivatives (OH-PBDEs and MeO-PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) in egg samples was developed by gel permeation chromatography ( GPC) and dispersive solid phase extraction ( DSPE) combined with liquid chromatography tandem mass spectrometric ( HPLC-MS/MS) and gas chroma-tography-negative chemical ionization mass spectrometry ( GC-NCI/MS ) . The analytes were extracted with mixture of hexane and dichloromethane (1∶1, V/V) by accelerated solvent extraction (ASE), and purified by 100 mg C18 dispersive solid phase extraction ( SPE) sorbents followed with gel permeation chromatography (GPC) , and then analyzed by liquid chromatography tandem mass spectrometric (HPLC-MS/MS) and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), respectively. The quantita-tion was carried out external standard method. The recoveries of objects were 64. 5%-97. 2% and 65. 6%-109 . 2% ( except BDE85 was 54 . 8%, OH-BDE-137 was 47 . 4%) spiked at 1 . 0 μg/kg or 5 . 0 μg/kg in egg white and egg yolk, respectively. The relative standard deviations (RSDs) were less than 20. 2%. The limits of quantitation (LOQ) for the object were 0. 01-0. 2 μg/kg.

Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS ( 1 μg/mL) and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay (ELISA), and the expression of miR-146a of cells was detected by using Real-Time PCR (TaqMan probe).Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student's t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge (P ＜ 0.01 ). The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells( P ＜0.01 ), and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.

Objective To develop a new method for the rapid determination of lomefloxacin,gatifloxacin,ciprofloxacin and oflox-acin in plasm and urine by solid phase extraction(SPE)and capillary electrophoresis.Methods The capillary was fused silica capil-lary with id/od of 75/365 μm and effective/total length of 40/47 cm.The running buffer was 40 mmol/L borate buffer at pH 9.0. Separation voltage was 13 kV.Temperature was 20 ℃.Detection wave-length was set at 280 nm.The sample was analyzed after the pretreatment of SPE.Results The analysis of lomefloxacin,gatifloxacin,ciprofloxacin and ofloxacin was completed in 6 minutes with satisfied accuracy and precision.Good linearity was found within the range of 1-40 μg/mL,and the r was 0.998 7,0.997 6, 0.998 3 and 0.994 2 respectively.The recoveries of four quinolones in plasm and urine ranged from 80.1% to 107.6%,and the rel-ative standard deviations(RSD)ranged from 2.1% to 6.2%.Conclusion This method is fast,simple,precise and it might be feasi-ble for the determination of lomefloxacin,gatifloxacin,ciprofloxacin and ofloxacin in plasm and urine samples.

Objective To investigate and analyze the capabilities of Plasmodium microscopic examinations in medical and health organizations in Dongxihu District,Wuhan City,so as to provide evidences for achieving the standard of malaria elimina?tion and malaria control in the future. Methods The basic information of the laboratorians and the hardware facilities in 15 mi?croscopic examination stations of Plasmodium were collected and analyzed from 2010 to 2015. Meanwhile,the quality of mak?ing,dyeing,cleanliness of all the negatives blood smears were evaluated,and all the positive smears were re?checked in the dis?trict during the period above mentioned. Results Totally,there were 28 laboratorians and 9 qualified microscopes in 15 micros?copy stations in Dongxihu District. The age of the laboratorians in the first?level and third?level organizations was mainly from 40 to 50 years old,but the age of those in the second?level organizations was from 20 to 30 years old. The years for microscopic ex?amination working of most of the laboratorians in the first?level and third?level organizations were more than 15 years,while the working years of those in the second?level organizations were mainly less than 10 years. The education level of the laboratorians in the first?level organization was mainly(50.00%)junior college,while the education levels of those in the second and third or?ganizations were mainly bachelor. From 2010 to 2015,8 561 blood smears were produced and 539 of them were reviewed,the co?incidence rates of review were all 100%,and the average qualified rates of making,dyeing,and cleanliness of the negative blood smears were 82.08%,79.92%and 83.33%,respectively. All the rates mentioned above in 2015 were higher than those in 2010(χ2=26.45,16.85,13.93,all P<0.01). Conclusion The capability of microscopic examinations of Plasmodium has improved obviously in Dongxihu District,but the human resources construction and hardware input in this district should be im?proved,meanwhile,the check on the quality control of microscopic examinations of Plasmodium should be continued,so as to guarantee the sustainable and healthy development of malaria control work in this district.

ObjectiveTo observe the effect of acetylcholine (ACh) on lipopolysaccharide (LPS) induced inflammatory model of rat alveolar macrophages, and to observe the effect of the acetylcholinesterase inhibitor physostigmine (Phy) on the anti-inflammatory effect of ACh.Methods The rat alveolar macrophages NR8383 were cultured in vitro, which were divided into five groups: blank control group, LPS group (stimulated with 1 mg/L LPS for 12 hours), LPS+ ACh group (0.01, 0.1, 1, 10, 100μmol/L of ACh were added for 5 minutes before LPS stimulation), LPS+ Phy group (1 mmol/L Phy was added for 5 minutes before LPS stimulation), and LPS+ ACh+ Phy group (1 mmol/L Phy and 10μmol/L ACh were added for 5 minutes before LPS stimulation). The supernatants were collected in each group, the enzyme-linked immunosorbent assay (ELISA) was used to assay the contents of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, and IL-6). The activity of acetylcholine esterase (AChE ) in the supernatant was also determined.Results① The contents of TNF-α (ng/L: 605.09±57.13 vs. 34.07±8.62), IL-1β (ng/L: 377.09±28.55 vs. 32.33±10.62) and IL-6 (ng/L: 558.04±77.45 vs. 42.62±11.21) in the LPS group were significantly higher than those in the blank control group (allP< 0.05). These results indicated that the inflammatory model of rat alveolar macrophages was constructed successfully.② ACh with the final concentrations of 0.01, 0.1, and 1μmol/L had less influence on the production of TNF-α, IL-1β and IL-6 in the culture supernatants of alveolar macrophages stimulated with LPS compared with LPS group (allP> 0.05). Nevertheless, 10μmol/L and 100μmol/L ACh notably reduced the production of TNF-α (ng/L: 451.19±30.67, 332.19±32.19 vs. 604.96±22.56), IL-1β(ng/L: 261.08±24.78, 143.98±28.39 vs. 367.06±10.44) and IL-6 (ng/L: 342.75±54.60, 235.48±29.75 vs. 562.69±63.34) in the culture supernatants compared with the LPS group (allP< 0.05).③ The activity of AChE in the LPS group was significantly higher than that in the blank control group (kU/L: 5.21±0.63 vs. 3.09±0.10,P< 0.05). The activity of AChE was successfully inhibited by 1 mmol/L acetylcholinesterase inhibitor Phy pretreatment compared with that in the LPS group (1.51±0.12 vs. 5.21±0.63,P< 0.05).④ The level of TNF-α (ng/L: 183.17±35.44 vs. 451.19±30.67), IL-1β (ng/L: 91.49±12.27 vs. 261.08±24.78) and IL-6 (ng/L: 108.17±22.82 vs. 342.75±54.60) in the culture supernatants of LPS+ ACh+ Phy group was significantly decreased as compared with LPS+ ACh group (allP< 0.05).Conclusions ACh with the final concentrations of 10μmol/L and 100μmol/L can inhibit the LPS induced inflammatory reaction in alveolar macrophages. The acetylcholinesterase inhibitor Phy can reinforce the ACh-mediated anti-inflammatory effect on alveolar macrophages inflammatory model.

Objective:To investigate the effect and mechanism of exosomes derived from human-induced pluripotent mesenchymal stem cells (iMSC-Exos) on alveolar macrophages (AM) pyroptosis.Methods:The exosomes in the culture supernatant of human-induced pluripotent mesenchymal stem cells (iMSC) were extracted by rotating ultrafiltration, and the extracted exosomes were identified by transmission electron microscopy, Western blotting and high-resolution adjustable resistance pulse. The rat alveolar macrophage cells (NR8383 cells) were cultured in vitro and the logarithmic growth phase cells were divided into three groups: the control group was added with an equal volume of phosphate buffered saline (PBS) in the AM supernatant; in LPS/ATP group AM cells were stimulated with 500 μg/L LPS for 23 hours and then 5 mmol/L ATP was added for 1 hour to induce pyrolysis; iMSC-Exos group was incubated with AM and 100 mg/L iMSC-Exos for 3 hours before giving LPS and ATP. The cytotoxic activity was detected by cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) analysis, the apoptosis and the expression of caspase-1 were observed by immunofluorescence, the levels of inflammatory factors interleukins (IL-1β and IL-18) released by AM were detected by enzyme linked immunosorbent assay (ELISA), the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the expression level of pyroptosis related protein gasdermin D (GSDMD) were detected by Western blotting. Results:The extracted exosomes were observed by transmission electron microscopy as round vesicles, expressing exosomal markers CD63 and CD9 showed by Western blotting, high-resolution adjustable resistance pulse showed the average diameter of the particles was 130 nm, and could be uptaken by AM. Compared with the control group, the cell activity decreased [(0.56±0.05)% vs. (1.06±0.07)%, P < 0.01], the release of necrotic substance LDH increased (U/L: 1 218.86±22.73 vs. 188.30±1.61, P < 0.01), the expression levels of inflammatory factors increased [IL-1β (ng/L): 958.91±32.78 vs. 194.63±5.14, IL-18 (ng/L): 870.89±21.86 vs. 288.85±24.48, both P < 0.01], and the apoptosis rate [(55.35±6.19)% vs. (12.01±1.32)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 41.06±3.65 vs. 2.80±0.54, P < 0.01) elevated in the AM after LPS/ATP stimulation, suggesting that LPS combined with ATP successfully induced alveolar pyroptosis. Compared with the LPS/ATP group, AM pretreated with iMSC-Exos showed increased cell viability [(0.81±0.05)% vs. (0.56±0.05)%, P < 0.01], decreased LDH secretion (U/L: 535.05±42.55 vs. 1 218.86±22.73, P < 0.01), decreased expression of inflammatory factors [IL-1β (ng/L): 381.82±19.50 vs. 958.91±32.78, IL-18 (ng/L): 533.77±31.54 vs. 870.89±21.86, both P < 0.01], and decreased apoptosis rate [(19.74±2.96)% vs. (55.35±6.19)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 12.16±1.31 vs. 41.06±3.65, P < 0.01). At the same time, the expression of NLRP3 inflammasome pathway [NLRP3 protein (NLRP3/β-actin): 0.62±0.06 vs. 1.89±0.11; cleaved caspase-1 protein (cleaved caspase-1/β-actin): 0.42±0.07 vs. 1.22±0.17, both P < 0.01] and pyrolysis-related protein was significantly inhibited [GSDMD protein (GSDMD/β-actin): 0.57±0.05 vs. 1.22±0.05, P < 0.01]. Conclusion:iMSC-Exos successfully reversed the AM pyroptosis and inflammatory factor expression induced by LPS/ATP, which may be due to the targeted inhibition of NLRP3 inflammasome pathway, suggesting that iMSC-Exos can exert anti-inflammatory effects by inhibiting the pyrolysis of AM.

AIM:To investigate the role of microRNA-132 (miR-132) on alveolar macrophage inflammation. METHODS: Rat alveolar macrophage cell line NR8383 was transfected with miR-132 mimic, mimic negative control ( NC) , miR-132 inhibitor, or inhibitor NC.The cells were divided into transfection group, transfection +lipopolysaccha-ride ( LPS) group, and transfection +LPS +acetylcholine ( ACh) group.The mRNA expression of acetylcholinesterase ( AChE) was detected by real-time PCR.The protein levels of AChE, signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) in the cells, and nuclear factor-κB (NF-κB) in the cytoplasm and nu-cleus were analyzed by Western blot.The activity of AChE in the culture supernatant was measured by AChE activity assay kit.The nuclear translocation of NF-κB was detected by immunofluorescence assay.RESULTS: Up-regulation or down-regulation of miR-132 had no effect on the mRNA expression of AChE.However, up-regulation of miR-132 decreased the protein level of AChE compared with mimic NC group (P<0.05).Transfection with miR-132 inhibitor increased the pro-tein expression of AChE compared with inhibitor NC group ( P<0.05 ) .In the alveolar macrophages treated with LPS+ACh, the inhibition of nuclear translocation of NF-κB p65 in miR-132 mimic group was more effective than that in mimic NC group ( P<0.05) .The inhibitory effect in miR-132 inhibitor group was weaker than that in inhibitor NC group ( P<0.05 ) .The inhibitory effect of miR-132 mimic on the protein levels of STAT3 and p-STAT3 was stronger than that of mimic NC (P<0.05).CONCLUSION:miR-132 in LPS-stimulated alveolar macrophages reinforced ACh-mediated anti-inflam-matory reaction by targeting AChE to suppress ACh hydrolyzation, which was related to the suppression of NF-κB and STAT3 activation.

OBJECTIVETo investigate the curative effect of antiphlogistic agent series on treating chronic nonbacterial prostatitis (CNP).

METHODSOne hundred and sixty patients were randomized into 4 groups for an 8-week clinical observation: group A (oral antiphlogistic medicinal granules only), group B (oral antiphlogistic medicinal granules + retention enema), group C (oral antiphlogistic medicinal granules + rectal), and group D (antiphlogistic medicinal granules + rectally + hip bath). Single blind trials were employed.

RESULTSThe curative rates of the 4 groups were 37.5%, 57.5%, 52.5% and 82.5% respectively, while the total efficacy rates were 42.5%, 82.5%, 77.5% and 92.5% respectively. Compared with groups A, B and C, the curative rate of group D was significantly higher (P < 0.05). The difference in efficacy rates was slight between groups B and D (P < 0.05), but significant between groups A and C (P < 0.05).

CONCLUSIONSCombined treatment therapy can improve the effect of CNP treatment and clear away heat and toxic material. The antiphlogistic agent series, with the effect of motivating blood circulation and removing blood stasis, turned out to be an effective traditional Chinese medicine in treating CNP.

Adult ; Chronic Disease ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Prostatitis ; drug therapy

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.