The toad, known for its various medicinal properties including parotid gland secretion (toad venom), dried skin, and gallbladder (toad bile), holds considerable medicinal applications as a valuable traditional Chinese animal medicine. Currently, in-depth attentions have been paid to the chemical composition and pharmacological properties of toad venom and skin; however, a lesser number of detailed analyses were concentrated on the toad bile. This review provides an overview of the chemical constituents in the bile of the Bufo genus, with a special focus on the cholestane and bufadienolides, and highlights the progress in their biosynthetic pathway and pharmacological activities. The analysis uncovers a distinct category of unsaturated Δ²² or Δ²³-C₂₇/C₂₈ bile acids in the toad gallbladder, potentially acting as key intermediaries in forming C-17 α-pyrone of bufadienolides. Furthermore, the high presence of 3α-OH configured bufadienolides in toad bile, in contrast to the common 3β-OH configured found in toad venom or skin, indicates a possible link between their minimal toxicity and the toad's self-defensive or physiological control. This review provides scientific basis for the development and utilization of toad bile resources, and provides useful reference for the discovery of lead compounds, analysis of the biosynthetic pathway of bufadienolides, and research on toad physiology.

[Abstract] Objective To investigate the appropriate X-ray angle which image can represent total proximal of femur according to the anatomic shape of the femoral neck section. Methods The anatomic parameter of the femoral neck section was observed and measured. Found the femoral neck was contained with three main surfaces of anterior surface, posterosuperior surface and posteroinferior surface. A model of surface fluoroscopy has been made by covering aluminum foil papers on the three surfaces of femoral neck. Taking the midpoint of the femoral neck as the fluoroscopy center, the C-arm was rotated at an interval of 10° between 0° and 170°, and the three aluminum foil paper models were taken for X-ray projection respectively. The X-ray angle with the smallest image of aluminum foil paper was taken as the appropriate X-ray angle of the surface. A penetrating model of the femoral neck internal fixation was made of three Kirschner wires penetrating through the anterior, posterosuperior and posteroinferior surfaces of femoral neck. The images of the penetrating Kirschner wires were observed through the traditional anteroposterior, lateral view and the three appropriate X-ray angle views obtained from the above test. Results The length of the three main surfaces accounts for 80. 95% of the circumference of the femoral neck. The anterior surface coronal angle was 18°, the posterosuperior surface coronal angle was 65°, and the posteroinferior surface coronal angle was 165°. The Appropriate X-ray angle of the three surfaces were 20°, 70° and 170° respectively. In the images of the three appropriate X-ray angles, the penetrating Kirschner wires of anterior surface, posterosuperior surface and posteroinferior surface of the femoral neck could be clearly observed respectively, while the penetrating Kirschner wire could not be completely found in the traditional anteroposterior and lateral images. Conclusion Traditional anteroposterior and lateral fluoroscopy can not accurately display the three-dimensional structure of femoral neck. By increasing the appropriate X-ray angle of 20°, 70° and 170°, the reduction quality and bony violation from internal fixation implants on the anterior surface, posterosuperior surface and posteroinferior surface of the femoral neck could be observed respectively.

Objective To explore protein extraction efficiency from formaldehyde-fixed paraffin embedded (FFPE) esophageal squamous cell carcinoma (ESCC) tissue samples with different protocols. Methods Six different lysis buffers with 100 °C or 105 °C. treatments were used for protein extraction, followed by evaluation of protein quantity and quality with Bradford, sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis, Western blotting and immunohistochemistry (IHC), using 8 FFPE samples of ESCC. Results The optimal method for protein extraction from FFPE ESCC tissue was Laemmli lysis buffer (Buffer 4) treated with 100 °C incubation, evidenced by highest amount of protein recovery. Western blotting and IHC method measured consistent 14-3-3σ expression in FFPE ESCC tissue samples. Protein precipitated by two volumes of acetonitrite acetonitrile(ACN) (0.1% trifluoroacetic acid) relative to protein amount reduced background staining on SDS-PAGE gels by commassie staining. Conclusion Laemmli lysis buffer combined with 100 °C incubation has the highest protein extraction efficiency from FFPE ESCC tissue samples for Western blotting measurement of protein biomarkers, and ACN protein precipitation can further eliminate residual cross- linked protein by FFPE.

Andrographis Herba is a commonly used plant medicine, and has been recorded in pharmacopeias of different countries. However, there are some differences in the quality standards. Based on this, this paper compare the quality standards of Andrographis Herba between Chinese Pharmacopoeia, Hong Kong Chinese Materia Medica Standards, United States Pharmacopoeia, European Pharmacopoeia and Indian Pharmacopoeia, including origin, botanical characteristics, identification(microscopic identification and chromatographic identification), content determination, specific test(such as impurities, loss on drying, extractives, pesticides, heavy metals, mycotoxins, and other items) and storage requirements, so as to provide a reference for studying international quality standards of Andrographis.
Andrographis ; Drugs, Chinese Herbal ; Materia Medica ; Reference Standards

OBJECTIVETo evaluate the clinical effects of French door segmented laminectomy decompression for severe cervical OPLL complicated with spinal cord injury.

METHODSThe clinical data of 38 patients with serious cervical OPLL complicated with spinal cord injury were retrospectively analyzed and these patients were treated with French door segmented laminectomy decompression and internal fixation from June 2012 to June 2014. There were 25 males and 13 females, aged from 42 to 78 years with an average of 58.2 years. Of them, 35 cases suffered from aggravating neurological symptoms with a definite precipitating factor. Spinal cord injury was related to minor injury of the neck, such as hyperextension of the neck in 3 cases. Preoperative Japanese Orthopaedic Score (JOA) was 8.1±1.7 and Neck Disability Index (NDI) was 19.8±4.4. Preoperative CT scans showed the range of OPLL was more than three segments. The spinal canal was occupied 50% to 85% with an average of 70.7%.

RESULTSAll the patients were followed up for 10 to 24 months with an average of 15.6 months. The operative time was 90 to 150 min with an average of 120 min and blood loss was 300 to 800 ml with an average of (480±80) ml. At final follow-up, NDI and JOA were 7.5±2.5 and 13.5±2.0, respectively, and they were obviously improved compared with preoperation. Preoperative cervical Cobb angle was (8.10±2.70)° and at final follow-up was (15.60±1.80)°, and there was significant difference between preoperative and postoperative (<0.05). Deep infection occurred in 1 case, epidural hematoma in 1 case, C₅ nerve root palsy in 3 cases, and axial symptom in 8 cases after operation. No serious complications, such as vertebral artery injury, cerebrospinal fluid leakage, deterioration of neurological dysfunction, or internal fixation failure was found.

CONCLUSIONSFrench door segmented laminectomy decompression is safe and feasible for severe cervical OPLL complicated with spinal cord injury, and it is worth to be popularized in future.

Objective To evaluate the clinical effect of Xuefu Zhuyu decoction combined with western medicine on elderly patients with refractory hypertension.Methods A total of 120 elderly patients with refractory hypertension were randomly divided into treatment group (n =60) and control group (n =60).Control group was given propranolol 5-10 mg orally,tid or qid;captopril 12.5 mg orally bid or tid;indapamide 2.5 mg orally qd.Treatment group was given Xuefu Zhuyu decoction 100 mL · d-1,bid on the basis of control group.All patients were treated for 90 d.The clinical symptoms and blood pressure were compared before and after treatment.Results The clinical efficacy was 95.00％ (57/60 cases) and 78.33％ (47/60 cases,P ＜0.05).After treatment,the systolic blood pressure (SBP) were (130.09 ±9.95),(113.08 ±15.24) mmHg in control group and treatment group,and diastolic blood pressure (DBP) were (84.40 ±9.05),(81.02 ±6.56) mmHg (P ＜0.05).There was 1 case of dizziness in control group,and no adverse drag reaction occurred in treatment group,with no significant difference (P ＞ 0.05).Conclusion Xuefu Zhuyu decoction combined with western medicine maybe an effective treatment on elderly patients with refractory hypertension.

Accumulative evidences have underpinned the nature candidates from Chinese medicine (CM), particularly CM served as blood activating and stasis resolving (BASR, Huoxue Huayu in Chinese) by targeting tumor-associated angiogenesis. However, recent experiment research on the therapeutic angiogenesis by BASR-CM attracts wide attention and discussion. This opinion review focused on the underlying link between two indications and anticipated that (1) BASR-CM might emphasize on a balanced multi-cytokines network interaction; (2) BASR-CM might address on the nature of diseases prior to differently affecting physiological and pathological angiogenesis; (3) BASR-CM might mainly act on perivascular cells, either promotes arteriogenesis by increasing arteriogenic factors in ischemic diseases, or simultaneously keep a quiescent vasculature to impede angiogenesis in tumor context.
Angiogenesis Inhibitors ; chemistry ; therapeutic use ; Animals ; Antineoplastic Agents ; therapeutic use ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Humans ; Models, Biological ; Neovascularization, Pathologic ; blood ; drug therapy

OBJECTIVETo study the effect of aqueous extract of several kinds of herbs on human platelet aggregation and expression of P-selectin in vitro.

METHODSBlood was collected from volunteers. Effects of the prepared water extracts of herbs on platelet aggregation were monitored on a Packs-4 aggregometer. The fluorescence intensity of water extracts of Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae on the expression of P-selectin in human platelets of healthy persons was measured with flow cytometry.

RESULTSOut of several herbs investigated, Flos Carthami and Rhizoma Curcumae potently inhibited platelet aggregation after incubation with platelet-rich plasma (PRP) for 15 min. Caulis Spatholobi Flos Carthami and Rhizoma Curcumae inhibited adenosine-5'-diphosphate (ADP) or platelet activating factor (PAF)-induced platelet aggregation in PRP in a dose-dependent manner. In contrast to Flos Carthami and Rhizoma Curcumae, Caulis Spatholobi could not inhibit thrombin-induced platelet aggregation. Despite its inability to inhibit thrombin-induced platelet aggregation in PRP, Caulis Spatholobi had a greater anti-aggregating activity in PRP induced by ADP or PAF. Caulis Spatholobi and Flos Carthami showed significant inhibitory effects on the expression of P-selectin.

CONCLUSIONSCaulis Spatholobi, Flos Carthami and Rhizoma Curcumae have potent anti-platelet properties, and their inhibitory actions are mediated via different mechanisms. Caulis Spatholobi inhibited ADP-induced platelet aggregation but not by thrombin, indicating that its mechanism of action might be independent of the thromboxane pathway. The effect of Caulis Spatholobi and Flos Carthami were associated with suppressing the expression of P-selectin.

Adult ; Blood Platelets ; drug effects ; metabolism ; Curcuma ; chemistry ; Fabaceae ; chemistry ; Humans ; P-Selectin ; metabolism ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Function Tests ; Water ; chemistry ; Young Adult

H5 subtype avian influenza (AIV-H5) is a major causative agent of animalloimia a rapid and sensitive molecular biological diagnosis is crucial to the control program of AIV-H5. AIV-H5 real-time fluorescent reverse transcription loop-mediated isothermal amplification (qRT-LAMP) was established by means of heat treatment of the samples. The sensitivity, specificity and repeatability of this method were assessed and the performance of Calcein,SYBR Green I,HNB,SYTO 81 in colorimetric detection was comparatively analyzed to screen the optimum dye. The results showed the sensitivity of this method was 100 times higher than that of standard real-time fluorescent RT-PCR, and the detection limit was one copy of the gene per reaction. This method had no cross-reactivity with other common avian respiratory tract infectious disease-related pathogens such as IBV and NDV. The present study suggested Calcein was the optimum dye. Small-scale tests suggested this method was reliable for survey monitoring of AIV-H5 on the spot, indicating its potential applications in field investigation.
Animals ; Chickens ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; diagnosis ; virology ; Poultry Diseases ; diagnosis ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods ; Sensitivity and Specificity

OBJECTIVETo investigate the expression of suppressor of cytokine signaling(SOCS)-3 and caspase-3 and their correlative significance in endometriosis.

METHODSImmunohistochemical EnVision method was used to detect the SOCS-3 and caspase-3 protein expression in ectopic and eutopic endometrium (n = 32) of patients with endometriosis, as well as normal endometrium (n = 30) of women without endometriosis.

RESULTSSOCS-3 and caspase-3 proteins were expressed in all three groups and not affected by the menstrual cycles. The expression of SOCS-3 in ectopic endometrium (5.54 ± 2.12) was significantly lower than that in eutopic (7.39 ± 1.09, P = 0.001) and control group (7.48 ± 1.26, P < 0.01), but without difference between the eutopic and control group (P = 0.756). SOCS-3 expression in ectopic and eutopic endometrium was significantly lower in III/IV stages than that in I/II stages of endometriosis (P < 0.05). Significantly lower expression of caspase-3 protein was found in ectopic (3.20 ± 1.24) and eutopic endometrium (3.88 ± 1.93) as compared with the control group (6.49 ± 1.85, P < 0.01), however ectopic and eutopic endometrium showed no significant difference (t = 1.66, P = 0.10). There was no significant difference of the expression of caspase-3 in ectopic and eutopic endometrium at different disease stages (P > 0.05). Positive correlation was found between the expression of SOCS-3 and caspase-3 proteins in ectopic endometrium (r = 0.655, P < 0.01).

CONCLUSIONSOCS-3 may be involved in the development of endometriosis through inhibition of apoptosis of ectopic endometrial cells.

Adult ; Caspase 3 ; metabolism ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Menstrual Cycle ; Middle Aged ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism ; Uterine Diseases ; metabolism ; pathology ; Young Adult