[Effect of electroacupuncture on behavior and cell apoptosis of brain tissue in traumatic brain injury rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on behavior, pathomorphology and brain cell apoptosis in traumatic brain injury (TBI) rats, so as to explore its mechanisms underlying treatment of TBI. METHODS: Male SD rats were randomly divided into control (n=10), sham (n=30), model (n=30) and EA (n=30) groups, the latter three groups were further divided into 3, 7, 14 d subgroups （n=10）. TBI model was established by using a free fall brain injury striking device after exposing the local cranial bone (to induce the left parietal cerebral contusion). Twenty-four hours after modeling, EA (2 Hz, 1 mA) was applied at "Quchi"(LI11), "Neiguan"(PC6), "Zusanli"(ST36) and "Yongquan" (KI1) combined with acupuncture at "Shuigou"(GV26) and "Baihui"(GV20) for 15 min, once a day for 14 consecutive days. After 3, 7, and 14 days treatment, the behavioral function (balance, walking, nerve, and limb retraction) of the rats was evaluated. The histopathological changes of the injured brain tissue were observed by HE staining. TUNEL method was used to detect the apoptosis of cells in the brain injury area. RESULTS: After modeling, the scores of balance and walking in the model group were higher than those in the sham group (P<0.01, P<0.05), the neurological function score and the right limb retraction force were lower than those in the sham group(P<0.01). After 3 days treatment, the neurological function score and the right limb retraction force in the EA group were higher than those in the model group (P<0.05). After 7 and 14 days treatment, the scores of balance and walking function in the EA group were lower than those in the model group (P<0.05, P<0.01), while the scores of nerve function and right limb retraction force were higher (P<0.05, P<0.01). HE staining showed that modeling induced pathological changes such as the inflammatory cell infiltration, interstitial edema, necrosis, nuclear pyknosis and nuclear lysis were relatively milder in the EA group on the 7th and 14th day. Compared with the sham group, the cell apoptosis of brain injury area was higher in the model group (P<0.01); while it was lower on day 7 and 14 in the EA group in comparison with the model group (P<0.01, P<0.05). CONCLUSION: EA can improve the behavioral function, reduce the apoptosis of brain cells in the injured area, and promote the rehabilitation of craniocerebral injury.

[Analysis on the rule of clinical acupoint selection in treatment of puerperal insufficient lactation with acupuncture and moxibustion].

The rule and characteristics of clinical acupoint selection were explored in treatment of puerperal insufficient lactation with acupuncture and moxibustion. The clinical articles on the treatment of puerperal insufficient lactation with acupuncture and moxibustion were retrieved from the databases of CNKI, VIP and Wanfang from the date of establishment to June 1, 2019. The articles were screened in accordance with the inclusion and exclusion criteria. The prescriptions of acupuncture and moxibustion were extracted and normalized. Using Microsoft Excel 2016 software, the use frequency, meridians involved and acupoint distributions were analyzed. Using SPSS Statistics 21.0 software, the cluster and factor analyses were conducted. Totally, 102 articles were included,108 acupoint prescriptions were extracted, 65 acupoitns were designed and the total use frequency was 654 times. The top three acupoints used in treatment of puerperal insufficient lactation were Danzhong (CV 17), Rugen (ST 18) and Shaoze (SI 1). The most frequently involved meridians were the stomach meridian, the conception vessel, the small intestine meridian and the liver meridian. The acupoints were mainly distributed in the chest and four extremities. It was showed in cluster analysis that Rugen (ST 18), Shaoze (SI 1), Zusanli (ST 36) and Danzhong (CV 17) represent 3 clusters and a total of 7 common factors were extracted. The acupoint selection is based on syndrome differentiation in treatment of puerperal insufficient lactation with acupuncture and moxibustion, of which, the local acupoints are predominated and the distal acupoints are combined.

[Electroacupuncture Intervention Reduces Mammary Gland Hyperplasia by Regulating Serum Estradiol and Progesterone Levels and Their Mammary Receptor Expression in Female Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on hyperplastic mammary glands, serum estradiol (E2) and progesterone (P) contents, estrogen receptor alpha (ERα) and progesterone receptor (PR) expression of breast tissues in mammary gland hyperplasia (MGH) rats, so as to reveal its mechanisms underlying improvement of MGH. METHODS: Sixty female SD rats were randomly divided into control, model, EA, EA＋ovariectomy(OVX) and EA＋sham-OVX groups (n＝12 in each). The MGH model was established by injection of Estradiol Benzoate injection (0.5 mg/kg, once daily for 25 d) and P injection (0.5 mg/kg, once daily for 5 d after estradiol injection) into the medial hind-leg muscle. After model establishment, bilateral OVX was performed for rats of the EA＋OVX group, and sham OVX (only exposure of ovaries) was performed for rats of the EA＋sham OVX group. EA (2 Hz, 2 mA) was applied to bilateral "Tianzong"(SI 11), "Ganshu"(BL 18), "Zusanli" (ST 36) for point group A, and "Wuyi"(ST 15), "Hegu"(LI 4), "Danzhong" (CV 17) for point group B for 20 min, once daily, 5 days a week for 4 weeks. The two acupoint groups were used alternately. The height of the rats' nipples (the 2nd pairs) were measured. Serum E2 and P levels were assayed by ELISA. The expression of ERα and PR in mammary tissues was detected by immunofluorescence staining and Western blot, separately. RESULTS: The height of nipples, serum E2content, and mammary ERα protein expression level were significantly higher in the model group than in the control group (P<0.01), while serum P content and PR expression were remarkably lower in the model group than in the control group (P<0.01). Following EA intervention, the height of nipples, serum E2content and mammary ERα protein expression level in both EA and EA＋sham OVX groups, and ERα expression in the EA＋OVX group were considerably lower in comparison with those of the model group(P<0.01), and serum P content and PR expression in the 3 EA groups were significantly higher than those in the model group (P<0.01，P<0.05). But there were no significant changes in the nipple height and serum E2 levels in the EA＋OVX group relevant to the model group (P>0.05), suggesting an important role of the ovary in producing EA's effect. CONCLUSION: EA intervention can improve hyperplastic mammary glands by down-regulating serum E2content and mammary ERα protein expression, and up-regulating serum P content and PR expression in MGH rats, which has a close relation with the intact ovaries.

Simultaneous determination of scopoletin, psoralen, bergapten, xanthotoxin, columbianetin acetate, imperatorin, osthole and isoimperatorin in rat plasma by LC-MS/MS for pharmacokinetic studies following oral administration of Radix Angelicae Pubescentis extract.

A rapid and sensitive bioassay based on liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed and validated for the simultaneous determination of eight coumarins in rat plasma. The liquid-liquid extraction method with ethyl acetate was used to prepare the plasma samples after addition of warfarin as an internal standard (IS). Chromatographic separation was performed on an Eclipse plus C18 column (100mm×4.6mm, 1.8µm) using gradient elution when 1mM ammonium acetate aqueous solution - acetonitrile was used as the mobile phase. The lower limit of quantitation (LLOQ) of each coumarin was lower than 2.16ngmL(-1). Intra-day and inter-day precisions were less than 15%. The accuracies were in the range of 88.9-117%. The mean recoveries of coumarins and IS were higher than 84%. The method was successfully applied to a pharmacokinetic study of eight coumarins in rats after oral administration of radix angelicae pubescentis.

The experimental study of Astragalus membranaceus on meridian tropsim: the distribution study of astragaloside IV in rat tissues.

According to Traditional Chinese Medicine (TCM) theories, TCM with different meridian tropism have different therapeutic effects. In view of the meridian tropism of Astragalus membranaceus (Huangqi), astragaloside IV, one of the effective phytochemicals of Huangqi, was appointed and observed its distribution in rat tissues following a single intravenous (i.v.) dose. A simple and accurate LC-ESI-MS/MS method was developed and validated for astragaloside IV quantification in heart, liver, spleen, lung and kidney using warfarin as an internal standard (IS). Chromatographic separation was performed on a Eclipse plus C18 (4.6mm×100mm, 1.8µm) when the flow rate was set at 0.300mLmin(-1) and ammonium acetate aqueous solution - acetonitrile was used as mobile phase. The intra- and inter-day precisions of the quality control samples were within 15% and accuracies were within 90.0-110%. The recoveries were more than 90.0% at high, medium and low concentrations, respectively. This method was successfully applied for distribution of astragaloside IV after intravenous (i.v.) dose of 4mgkg(-1) astragaloside IV in rats. Astragaloside IV concentration was highest in liver and kidney and remained much higher than that in other tissues over the experiment course. Lung, heart and spleen were also detected to contain astragaloside IV. The results clearly demonstrated that astragaloside IV was one of the material bases of the meridian tropism of Huangqi.