Objective To study the action of puerarin on Caspase-3 and Bcl-2 expression of hippocampal CA1 neurons in ovariectomized rats, and explore its mechanism. Methods SD female rats were randomly assigned into sham operation group, model group, premarin group and puerarin groups (120, 60, 30 mg/kg). The model group and sham operation group were injected NS intraperitoneally, other groups were treated with corresping drugs for 30 d. Immunohistochemistry and RT-PCR were used to determine Caspase-3 and Bcl-2 expression of hippocampal CA1 neurons. Results Caspase-3 expression of hippocampal CA1 neurons was significantly decreased in high-dose puerarin group (P <0.05). Bcl-2 expression of hippocampal CA1 neurons was significantly increased in high- and medium-dose puerarin groups (P<0.05, P<0.01). Bcl-2 mRNA level of hippocampal CA1 neurons in high-dose puerarin group was significantly increased (P<0.05). Conclusion Puerarin can decrease Caspase-3expression and increase Bcl-2 expression of hippocampal CA1 neurons in ovariectomized rats, and has protective effect on neuronal structure.

BACKGROUND:Immortalized cervical epithelial cels H8 can become cancerous under the induction of carcinogenic agent, and may cause cervical cancer when there is a cofactor interaction. However, there is stil a lack of effective intervention for female patients with precancerous lesions, and this treatment is blank in the clinic. OBJECTIVE: To explore the effects and mechanism of astragalus injection on apoptosis of human immortalizedcervical epithelial cels H8. METHODS: This study contained two groups: astragalus drug group and the blank control group. (1) Enzyme linked immunosorbent assay (ELISA) was used to detect DNA fragments of apoptotic H8 after astragalus injection. (2) Enzyme-labeling instrument was used to analyze the changes in caspase-3 and caspase-9 activities a fter astragalus injection. (3) Western blot assay was used to detect the protein expression changes of caspase-3, caspase-9 and PARP in H8 cels after astragalus injection. RESULTS AND CONCLUSION: (1) ELISA results showed that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, DNA fragments were gradualy increased with time prolonged in a time-dependent effect (P < 0.05). (2) Enzyme-labeling instrument demonstrated that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, caspase-3 and caspase-9 activities increased in a time-dependent manner (P < 0.05). (3) At 0, 6, 12 and 24 hours after 20 g/L astragalus injection, the expression of cleaved caspase-3 and cleaved caspase-9 were gradualy increased in H8 cels (P < 0.05). Cleaved PARP protein expression was gradualy decreased (P < 0.05). These findings indicate that astragalus injection could obviously induce H8 apoptosis, which may be associated with the upregulated protein expression of caspase-3 and caspase-9.

Objective To discuss the effects of Radix Hedysari flavonoids on microvessel density (MVD) of pulmonary interstitial fibrosis model rats, and provide evidence for its development. Methods SPF Wistar rats were divided into blank group, model group, prednisone group, and Radix Hedysari flavonoids high-, medium- and low-dosage group. Pulmonary interstitial fibrosis model was established by intratracheal instillation of bleomycin. From the second day after model established, each drug treatment group was administered with corresponding drugs intragastrically at different points in time for 14 and 28 days. MVD was detected by immunohistochemistry. Results Neomicrovessel was increased significantly in pulmonary tissue of model rats of 14 days treatment, but slightly decreased in 28 days. MVD in 14, 28 days model group was significantly more than blank group (P<0.01). MVD in 14, 28 days Radix Hedysari flavonoids high dosage group was significantly less than model group (P<0.01). MVD in Radix Hedysari flavonoids medium-dosage group was between high-and low-dosage group, and MVD in medium-dosage group was similar with that in model group. Conclusion Radix Hedysari flavonoids could inhibit the formation of neomicrovessel in a dose dependent manner.

Botulinum toxin type A (BTX A) has gain widespread use in various neurological conditions, characterized by safe injection and less side-effect. This review will examine the current research of BTX A in the following aspects: mechanism, location, dose and outcome, which manifests the safety and efficacy data of BTX A injection treatment to the symptoms of Parkinson disease, including refractory tremor, focal dystonia, loss of postural reflexes, sialorrhea, gastrointestinal symptoms and depressive disorder.

Radiation-induced bystander effect (RIBE) refers to that irradiated cells release signaling factors and induce responses in nonirradiated cells.In other words, it is the communication between irradiated and nonirradiated cells by intracellular signals. RIBE could influence the efficacy of tumor radiotherapy, but also has potential risk to the normal tissues outside of radiation field. Studies have found that ionizing radiation can induce the alteration of miRNA expression not only in the irradiated cells but also in adjacent nonirradiated tissues, and miRNAs may play an important role in the regulation of signaling pathways between irradiated and nonirradiated bystander cells. This article reviewed the roles of miRNAs in RIBE.

OBJECTIVETo observe the effect of Pingxian granules on the protein expression of apoptosis regulatory genes Bcl-2 and Bax in the hippocampus of epileptic model rats and study the molecular biological mechanism of the anti-epileptic effect of Pingxian granules.

METHODTotally 60 45-days-old Wistar rats were selected and then randomly assigned into 5 groups: the normal control group, the model group, the positive control group, the Pingxian high dose group and the Pingxian low dose group, with 12 in each group. Except the normal control group, all the groups were intraperitoneally injected with 35 mg x kg(-1) pentylenetetrazol to establish the models of epilepsy. The Pingxian high dose (1.66 g x mL(-1)) and low dose (0.42 g x mL(-1)) groups were intragastrically infused with Pingxian granules 2 mL x d(-1). The positive control group received 3.6 g x L(-1) phenobarbital suspension by gastric perfusion. The normal group and the model group were drenched with distilled water, 2 mL x d(-1), for 5 weeks. Bcl-2 and Bax protein positive cells were labeled with immunohistochemical SABC at 3, 5 w.

RESULT(1) Rats in the model group appeared the epileptic behavior at the 1st week, and became serious with the kindle frequency; grade VIepileptic behavior appeared at the 4th week. The attack frequency and grade of the Pingxian group were less and lower, the highest grade were only IV, and there were no significant differences in the attack grade and frequency. (2) With the increase in kindle frequency, the model group showed a notable decrease in the Bcl-2 expression compared with the normal control group at the 3rd and 5th weekend (P < 0.01), but a significant increase in Bax protein expression (P <0. 01). The number of the Bcl-2 protein expression in Pingxian groups and the positive control group were obviously more than the model group (P < 0.01); and the number of the Bax protein expression in Pingxian groups and the positive control group were obviously less than the model group (P < 0.01).

CONCLUSIONPingxian granules may decrease neuronal cell apoptosis by improving the protein expression of apoptosis regulatory gene Bcl-2 and inhibiting the protein expression of apoptosis regulatory gene Bax with a view of anti-epilepsy.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Epilepsy ; chemically induced ; drug therapy ; Female ; Hippocampus ; drug effects ; metabolism ; Immunohistochemistry ; Male ; Pentylenetetrazole ; toxicity ; Rats ; Rats, Wistar

Objective To compare the cumulative analgesic effects of electroacupuncture at Sanyinjiao (SP6), Xuanzhong (GB39) and non-acupoint in treating primary dysmenorrhea. Method By adopting a multi-centered randomized controlled study method, 501 patients recruited from Dongzhimen Hospital of Beijing University of Chinese Medicine, China-Japan Friendship Hospital, Beijing Hospital of Traditional Chinese Medicine of Capital Medical University, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Huguosi Hospital of Chinese Medicine of Beijing University of Chinese Medicine and the Outpatient of Shandong University of Traditional Chinese Medicine were randomized into a Sanyinjiao group, a Xuanzhong group, and a non-acupoint group, 167 subjects in each group. The electroacupuncture intervention was applied when dysmenorrhea flared up and the Visual Analogue Scale (VAS) ≥40 mm, with frequency at 2/100 Hz and intensity during patient’s endurance, 30 min each time, once a day, and for successive 3 d. Before the first treatment, 30 min after the first treatment, and respectively prior to the second and third treatment, VAS was used to measure the pain intensity. Meanwhile, the Retrospective Symptom Scale (RSS-COX 2) was investigated before the first treatment, right after the removal of needles for the first treatment, before the second and third treatment. Result The decrease of VAS in Sanyinjiao group was more significant than that in Xuanzhong group and non-acupoint group (MD=﹣2.92 mm, P=0.028; MD=﹣3.47 mm, P=0.009), while there was no significant difference between Xuanzhong group and non-acupoint group (MD=﹣0.56 mm, P=0.674); there were no significant differences in comparing the RSS-COX2 total score among the three groups (P=0.086). Conclusion Sanyinjiao (SP6) can produce a more significant cumulative analgesic effect for primary dysmenorrhea patient than Xuanzhong and non-acupoint, and the effects of Xuanzhong and non-acupoit are equivalent.

ObjectiveTo explore the effects of modified Danggui Beimu Kushen pills on tumor growth and T-cell subsets in H22 hepatocellular carcinoma-bearing mice and to provide an experimental basis for the treatment of hepatocellular carcinoma with modified Danggui Beimu Kushen pills combined with immune checkpoint antibodies. MethodA H22 hepatocellular carcinoma-bearing mouse model was established. The modeled mice were randomized into model， cisplatin， low- （4 g·kg^-1·d^-1）， medium- （8 g·kg^-1·d^-1）， and high-dose （16 g·kg^-1·d^-1） modified Danggui Beimu Kushen pills groups. After continuous administration for 14 days， the mice were sacrificed on day 15. The tumor volume was measured on days 0， 4， 8， 12， 15 of drug administration. Tumors were weighed and thymus index and spleen index were calculated. Spleen lymphocytes were co-cultured with H22 hepatoma cells， and the tumor cell-killing rate was detected by the cell counting kit-8 （CCK-8）. Real-time polymerase chain reaction was carried to determine the mRNA levels of programmed cell death protein-1 （PD-1） and lymphocyte activation gene-3 （LAG-3） in spleen and tumor tissues. The number of CD4⁺ and CD8⁺ T cells and the expression of PD-1 and LAG-3 were detected by immunohistochemistry （IHC）. ResultOn day 8 of drug administration， tumor volumes in all treatment groups decreased compared with that in the model group. On day 15， both tumor volume and tumor weight were significantly lower in the treatment groups than in the model group （P<0.01）， with the cisplatin group showing the most pronounced reduction. Compared with the model and cisplatin groups， medium- and high-dose modified Danggui Beimu Kushen pills increased the thymus index （P<0.01）. Compared with the model group， all treatment groups showed increased spleen index （P<0.05， P<0.01）， with the cisplatin group showing the most significant increase. Compared with the model and cisplatin groups， all the groups of modified Danggui Beimu Kushen pills demonstrated increased number of CD4⁺ and CD8⁺ T cells and tumor cell-killing rate in the spleen and tumor tissues （P<0.01） and down-regulated mRNA and protein levels of LAG-3 （P<0.05， P<0.01）. The high-dose group of modified Danggui Beimu Kushen pills had lower mRNA level of PD-1 in the tumor tissue than the model and cisplatin groups （P<0.01）. ConclusionModified Danggui Beimu Kushen pills may promote the proliferation and tumor microenvironment infiltration of CD4⁺ and CD8⁺ T cells in H22 tumor-bearing mice by down-regulating LAG-3 expression， thereby improving T-cell immune activity and inhibiting tumor growth. This study provides an experimental basis for the combination of modified Danggui Beimu Kushen pills and immune checkpoint antibodies in the treatment of hepatocellular carcinoma.