Recently, the hepatotoxicity issue regarding to Psoraleae Fructus (PF) has attracted remarkable concerns, which highlights the urgent need to explore the toxicity attenuation method for PF. In this study, we proposed an alcohol soaking and water rinsing method for pre-processing PF based on the record in the classics - "Lei Gong Pao Zhi Lun", aiming to attenuate the potential hepatotoxicity of PF. The optimal pre-processing methods and parameters were investigated by U*12(10⁸) uniform design coupled with 3D-cultured human-derived liver organoids model and high-content imaging. The results showed that there were significant variations among the hepatotoxicity intensities of different pre-processed PF products. Four factors, including the concentration of alcohol, the ratio of material and alcohol in alcohol soaking, the time of alcohol soaking and the times of water rinsing, were found as independent significant factors (all P<0.01). The optimal pre-process parameters were further predicted and verified as follows: the alcohol concentration is 80%, the times of alcohol soaking is 3, the ratio of alcohol and material of alcohol soaking is 3, the time for alcohol soaking is 30 h, the ratio of water and material of water rinsing is 2, the times of water rinsing is 3, the time water rinsing is 12 h and the time of steaming is 5 h. This research demonstrated that the alcohol soaking and water rinsing method can effectively reduce the potential hepatotoxicity of PF. This method provides a reference for reducing the risk of PF liver injury from the perspective of Chinese medicinal materials pre-processing.

To investigate the effect of overexpression of microRNA-21-5p (miR-21-5p) on early apoptosis of type Ⅱalveolar epithelial cells (AECⅡ) in rats with hyperoxic acute lung injury (HALI). Methods The Sprague-Dawley (SD) rats were randomly divided into four groups: control group (CON group), hyperoxia group (H group), overexpression group (OE group) and empty vector group (EV group), with 20 rats in each group. HALI animal model was made by inhaling high concentration oxygen (oxygen concentration ≥90%); CON group was arranged to inhale room air. The miR-21-5p adeno-associated virus-6 (AAV-6) overexpression vectors or empty vectors were dripped into the lungs of OE group and EV group through tracheal tube, respectively. The hyperoxia model was prepared after 3 weeks of feeding. At 0, 24, 48 and 60 hours after making model, 5 rats were selected to detect lung injury parameters:oxygenation index (OI), respiratory index (RI), wet/dry ratio (W/D), pathological injury score of lung tissue; real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-21-5p in AECⅡ, and flow cytometry was used to detect the early apoptotic rate of AECⅡ. Results ① The lung injury parameters: in H group, the OI gradually decreased with time, but the RI, lung W/D ratio and pathological score increased gradually with time, the difference between CON group was statistically significant at 24 hours [OI (mmHg, 1 mmHg = 0.133 kPa):336.04±5.79 vs. 400.22±19.70, RI: 0.20±0.02 vs. 0.10±0.06, lung W/D ratio: 5.04±0.09 vs. 4.85±0.09, lung tissue pathological score: 0.13±0.01 vs. 0.07±0.01, all P < 0.05]. It indicated that HALI model could be successfully established by inhaling high concentration oxygen continuously. ② The expression of miR-21-5p: the miR-21-5p was gradually increased in H, OE and EV groups, and the expression of miR-21-5p was significantly higher than that in CON group at 24, 48 and 60 hours. Compared with H group, the expression of miR-21-5p was significantly increased further in OE group at 0, 24, 48 and 60 hours (2-ΔΔCt: 3.75±0.11 vs. 0.98±0.14, 3.98±0.12 vs. 1.18±0.13, 4.28±0.18 vs. 1.49±0.06, 4.66±0.12 vs. 1.80±0.12, all P < 0.05). ③ The early apoptosis of AECⅡ: the early apoptosis rate gradually increased with time in H, OE and EV groups, and the early apoptosis of AECⅡ was significantly higher than that in CON group at 24, 48 and 60 hours. Compared with H group, the early apoptosis rate was significantly decreased in OE group at 24, 48 and 60 hours [(1.22±0.63)% vs. (2.84±0.59)%, (5.76±0.18)% vs. (13.10±2.01)%, (29.48±0.48)% vs. (49.04±1.36)%, all P < 0.05]. ④ There was no significant difference in the expression of miR-21-5p and the early apoptosis of AECⅡ cells between EV group and H group at each time point. Conclusion Overexpression of miR-21-5p could inhibit the early apoptosis of AECⅡ in rats with HALI.

OBJECTIVE: To explore the clinical effect of acupuncture and the potential effect mechanism in patients with premature ovarian failure. METHODS: A total of 104 patients with premature ovarian failure were randomized into an acupuncture group and a western medication group, 52 cases in each one. In the western medication group, the conjugated estrogens tablets were prescribed for oral administration, 0.625 mg each time, once a day, consecutively for 21 days. On the 16th day of medication with conjugated estrogens tablets, the oral administration of medroxyprogesterone acetate tablets were supplemented, 10 mg each time, once a day, consecutively for 5 days, and then, these two kinds of western medication were discontinued for 1 week. A total of 3 cycles were required in treatment with 28 days as an artificial cycle. In the acupuncture group, acupuncture was applied. Two groups of acupoints were selected. The first group of acupoints were stimulated before ovulation and the acupoints were Guanyuan (CV 4), Guilai (ST 29), Taichong (LR 3), Taixi (KI 3), Xuehai (SP 10), Sanyinjiao (SP 6), Zigong (EX-CA 1), Yinlingquan (SP 9), Zusanli (ST 36), Shuidao (ST 28), Dahe (KI 12) and Tianshu (ST 25). The second group of acupoints were stimulated after ovulation and the acupoints included Ciliao (BL 32), Shiqizhui (EX-B 8), Ganshu (BL 18), Shenshu (BL 23), Geshu (BL 17) and Pishu (BL 20). The therapeutic effect was observed and compared in the patients between the two groups, as well as the expressions of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) and the levels of serum luteinizing hormone (LH), follicule stimulating hormone (FSH) and estradiol (E) before and after treatment. RESULTS: The total effective rate was 90.4% (47/52) in the acupuncture group, higher than 67.3% (35/62) in the western medication group (<0.05). After treatment, the expressions of IFN-γ and TNF-α in the acupuncture group were obviously lower than the western medication group (<0.05). Except for serum LH after treatment, at the end of treatment and in 30 days and 90 days after treatment, the levels of serum E in the acupuncture group were higher obviously than the western medication group and the levels of serum LH and FSH were lower obviously than the western medication group (all <0.05). CONCLUSION Acupuncture promotes the regular menstruation, effectively regulates the levels of serum LH, FSH and E and improves the pituitary gland and the ovary endocrine in the patients with premature ovarian failure. Such effect may be related to the the improvements in the expressions of IFN-γ and TNF-α, the inhibition of the apoptosis of ovarian granulosa cells, the recovery of ovarian function and the enhancement of reserve capacity.
Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Interferon-gamma ; blood ; Primary Ovarian Insufficiency ; blood ; therapy ; Tumor Necrosis Factor-alpha ; blood

Objective: To intensively study the chemical constituents from the seed cake of Camellia oleifera and its pharmacological activities,in order to provide scientific basic for its further development and utilization. Method: All kinds of column chromatography and spectral methods were employed to isolate and identify the monomeric compounds from its ethyl acetate portion of ethanol extract. The in vitro anti-inflammatory effects were evaluated by LPS-induced inflammatory model in RAW264.7 macrophages. Result: Eight phenolic acids and two flavonoids were isolated from the ethyl acetate soluble portion and identified as p-hydroxybenzoic acid(1),protocatechuic acid(2),gallic acid(3),methyl gallate(4),ethyl gallate(5),isovanillic acid(6),ethyl 3,4-dihydroxylbenzoate(7),2-(3',4'-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde(8),quercetin(9),rutoside(10). Among them, compounds 4-8 were first isolated from this plant. These compounds had good anti-inflammatory activities against NO production in LPS-stimulated RAW264.7 macrophages in an obvious dose-dependent manner. Among them, compound 8 showed a strongest activity. Conclusion: The above results show that the phenolic acids and flavonoids from seed cake of C. oleifera have good prospects for the development and application of anti-inflammatory drugs.

Objective:To explore the effect of Zhizhuwan on intestinal conduction and expressions of Phospholipase C-γ₁ (PLC-γ₁)/Phospholipase C-γ₂ (PLC-γ₂) signaling pathway of slow transit constipation (STC) with spleen deficiency syndrome. Method:Special pathogen free (SPF) healthy mice were randomly divided into normal group and model making group. Folium Sennae gavage was used to induce the spleen deficiency status, and then diet and drinking water were controlled to establish the mice model of spleen deficiency constipation. After the modeling, the mice in modelling group were randomly divided into model group, Zhizhuwan group and mosapride group. Zhizhuwan group was given drug at the dose of 9.0 g·kg^-1·d^-1, mosapride group was given 2.5 mg·kg^-1·d^-1, model group and normal group were given the equal dose of distilled water for 7 consecutive days. At the end of the treatment, the length of Indian ink in the colon was used to calculate the intestinal propulsion rate of the mice. The D-xylose kit was used to determine the content of D-xylose in serum of mice. The hematoxylin eosin (HE) staining was used to observe the pathological changes of colon tissues in mice. The immunohistochemistry and Western blot were used to detect the expression levels of PLC-γ₁ and PLC-γ₂ proteins in colon tissues of mice. Real-time PCR was used to detect the mRNA expression levels of PLC-γ₁ and PLC-γ₂. Result:Compared with normal group, the intestinal propulsive rate and the serum D-xylose value in model group were significantly decreased (P<0.01). Obvious hyperemia and edema of colonic mucosa were observed by HE staining. The protein and mRNA expression levels of PLC-γ₁ and PLC-γ₂ in colon tissues were significantly decreased (P<0.01). Compared with model group, the intestinal propulsion rate and the content of D-xylose in Zhizhuwan group and mosapride group were significantly increased (P<0.01). Colonic mucosal hyperemia, edema and inflammatory cell infiltration were improved according to HE staining. The protein and mRNA expression levels of PLC-γ₁ and PLC-γ₂ were significantly increased (P<0.01). Conclusion:Zhizhuwan can promote the intestinal movement in slow transit constipation model mouse with spleen deficiency syndrome, and alleviate the symptoms of constipation in mice. The related mechanism may be related to the increase of the expressions of PLC-γ₁ and PLC-γ₂ in colon tissues of mice with spleen deficiency and constipation.

Objective: To explore the patient and hospital related determinants of adherence to early antithrombotic therapy among patients with acute ischemic stroke (AIS). Methods: AIS patients aged 50 years old or above who were eligible for early antithrombotic therapy, were included from the China National Stroke Registry Ⅱ (CNSR Ⅱ) project. Characteristics related to patients and hospitals were collected. Univariate analysis method was conducted to explore the correlation between hospital or patient-related determinants and early antithrombotic therapy. A 2-level logistic regression model was set up to identify patient and hospital-related variables that were associated with the adherence to early antithrombotic therapy, with patient as level 1 and hospital as level 2. Results: A total of 16 910 patients were included in the study, with 14 332 (84.75%) of them having received early antithrombotic therapy. Results from the univariate analysis showed that the patient determinants to early antithrombotic therapy would include age, type of health insurance, average income and history of dyslipidemia. Hospital determinants would include factors as: level and region of the hospital, academic status, with/without stroke unit, quality control on single disease and the percentage of neurological beds in total beds (P<0.05). Data on multilevel model showed that the patient-related determinants on early antithrombotic therapy would include age, gender, average income, history of hypertension, National Institutes of Health Stroke Scale (NIHSS) score at admission while hospital related determinants would include percentage of neurological beds in total beds, and region of the hospital (P<0.05). Conclusions The quality of a hospital was associated with the adherence to early antithrombotic therapy. AIS patients at advanced age or with high NIHSS score at admission should be paid more attention.

Objective To study the effects of fluorocitrate (FC),astrocytic metabolic inhibitor,on early brain injury after subarachnoid hemorrhage (SAH)in rats so as to explore the mechanism of early brain injury mediated by astrocyte after SAH.Methods Thirty six SD male rats were randomly divided into sham group,SAH 3 d group and SAH 3 d+FC group,with 12 rats in each group.The model of SAH was established by an endovascular perforation technique. FC was injected into the lateral ventricle. Three days after SAH, the expressions of GFAP,Iba-1 and TNF-α were detected using immunohistochemistry in all rats.The expression and phosphorylation of NF-κB in the nucleus were detected by Western blot.The expressions of TNF-α,IL-1βand IL-6 were detected by ELISA.Cell apoptosis was detected by TUNEL.Results Neuronal swelling,nuclear anomalies, disorganization of micrangium,abnormality of endothelial cells appeared in SAH 3 d group,but not in sham group. The expressions of NSE,GFAP and Iba-1 were significantly increased in SAH 3 d group compared with those in sham group (all P＜0.05).The number of apoptotic cells was increased.The expression and phosphorylation of NF-κB were significantly increased.The expressions of TNF-α,IL-1β and IL-6 were significantly increased.However, neuronal injuries were alleviated by injecting FC.The expressions of NSE,GFAP and Iba-1 in SAH 3 d+FC group were inhibited.The number of apoptotic cells in SAH 3 d+FC group was decreased compared with that in SAH 3 d group.The expression and phosphorylation of NF-κB were decreased by FC.The expressions of TNF-α,IL-1β and IL-6 were significantly decreased by FC (P＜0.05).Conclusion Astrocytes may play an important role in early brain injury after SAH,underlying the mechanism that they released TNF-α,IL-1βand IL-6 and activate microglia by activating NF-κB signal.The inhibition of excess activation of astrocytes may plays a protective role in early brain injury after SAH.

A simple, sensitive and reliable method was developed for simultaneous determination of ten banned drugs residues including zeranols(ZALs),chloroamphenicol,pentachlorophenol,etc. in swine urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The urine samples were pretreated using lyophilization and QuEChERS procedures, respectively. Acetonitrile and ammonium acetate (5 mmol/L) were chosen as mobile phases. Target compounds were separated well in ZorbaxSB-C18by following the optimized gradient elution program and determined by LC-MS/MS in negative electrospray ionization mode. The linearity of the matrix-matched standard curve of ten analytes in two methods was good in the range of the experimental concentration with correlation coefficients more than 0.99. The recoveries of ten drugs were in the range of 80.7%-107.7% and 73.5%-103.3% at the spiked levels of 5,10 and 20 μg/L by lyophilization and QuEChERS methods,respectively. The coefficients of variation were less than 15%. The limits of detection (LOD) and the limits of quantification (LOQ) from lyophilization and QuEChERS method were 0.1 to 2.0 μg/L and 0.2 to 5.0 μg/L,respectively.

A method was established for the determination of chlorpyrifos metabolites employing QuEChERS method and ultra performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry (UPLC-QTRAP). The urine samples were extracted by acetonitrile and then cleaned up with PSA and GCB. The samples were separated with the gradient elution of acetonitrile-0.2% ammonia water on ZORBAX Eclipse Plus C18column. The analytes were detected by tandem mass spectrometry under negative ion mode with electrospray ionization (ESI) source and MRM-IDA-EPI mode. Under the optimized conditions, the calibration curve was linear in range of 1.0-100.0 μg/L,and the limits of detection were 0.10-0.73 μg/L. The average recoveries were 80.3%-90.1%, and RSDs were all within 10%. The developed method was simple,sensitive,accurate,and repeatable,and could avoid false positive result of samples effectively. The established method was successfully applied to determine the exposure level of chlorpyrifos metabolites in real samples of human health risk analysis. The results showed that the maximum concentration of chlorpyrifos metabolites was 54.6 μg/L. This method provided technic support for simultaneous identification and quantification of chemicals in complex matrix.

Objective:This article analyzes the cognition and satisfaction evaluation of patients in medical alli-ance. Under the medical alliance mode,the present study tries to explore the choice of medical treatment of patients, the first diagnosis of primary health care institutions and its influencing factors, in order to provide reference for the construction of medical alliance. Methods:With the help of stratified sampling method,eighteen (18) medical alli-ances were selected from 16 districts and counties in Beijing. The survey questionnaire was designed to be used as the tool for data collection. A total of 1 280 patients in medical alliance participated in this study. The Chi-square Test and binary logistic regression model were used for statistical analysis. Results:59.12% of patients understand the medical association and related policies. 39.12% of the patients are satisfied with the medical association,51.44% of patients are willing to take first diagnosis at primary health care institutions. The distribution of medical resources in various districts is uneven. The type of hospital visited,the degree of understanding of medical alliance,the expe-rience of medical alliance and the satisfaction toward medical alliance will have an impact on the willingness about basic initial diagnosis,and the influence make a significant statistical sense. Conclusions:The Medical alliance con-struction in Beijing has achieved a certain effect,as first visit at primary health care institutions will be the initial for-mation. But the unbalanced distribution of medical resources in various districts,the service capacity of primary care institutions which does not meet the needs of patients,the insufficient sensitization of relevant policy and the current health insurance policy are seen as limiting factors to the development of medical alliance.