Natural products (NPs) are an important source of new drugs for the treatment of stroke. Identifying cellular targets for bioactive molecules is a major challenge and critical issue in the development of new drugs for stroke. Small-molecule probes play a unique role in target discovery. However, drawbacks to these probes include non-specificity, unstable activity, and difficulty in synthesis. Small-molecule probes based on NPs at least partially compensate for these shortcomings. NPs feature rich chemical and structural diversity, biocompatibility, and unique biological activities. These features could be exploited to provide new ideas and tools for target discovery. Small-molecule probes based on NPs provide a precise and direct search for interacting protein targets of NPs-active small molecules. This review explores the properties of small-molecule probes based on NPs and their applications in mechanistic studies of stroke and other diseases. We hope that this review will bring new perspectives to the mechanistic study of NPs-active small molecules and accelerate the translation of these ingredients into drug candidates for the treatment of stroke.

OBJECTIVETo study the inhibitory effect of CD133 monoclonal antibody labeled with ¹³¹I (¹³¹I-CD133mAb) on Huh-7 human liver cancer cell line overexpressing CD133 antigen in vitro and in mouse models bearing the tumor cell xenograft.

METHODS¹³¹I-CD133mAb was prepared by chloramines-T method and evaluated for its stability. Flow cytometry and immunohistochemistry were used to detect the expression of CD133 in Huh-7 cells and in Huh-7 cell-derived tumors, respectively. Huh-7 cells treated with ¹³¹I-CD133mAb plus cisplatin (DDP), ¹³¹I -CD133mAb, DDP, or no treatment (blank control) were examined for cell proliferation suppression by MTT assay with the IC₅₀ calculated. BALB/c mice bearing subcutaneous Huh-7 cell xenograft in the right forelegs were treated with ¹³¹I -CD133mAb, DDP, or both every two days for two weeks. The tumor size and volume were measured twice a week, and pathological examination of the tumor was carried out after the treatments. The tumor inhibition rate was calculated and tumor cell apoptosis observed with HE staining.

RESULTSThe labeling ratio of ¹³¹I-CD133mAb was 90.25% and the radiochemical purity was 97.78%. Huh-7 cells showed obviously higher CD133 expression than HepG2 cells. ¹³¹I-CD133mAb combined with DDP group resulted in a significantly higher tumor inhibition rate than other treatments in the tumor-bearing mice.

CONCLUSION¹³¹I-CD133mAb can inhibit the growth of liver cancer cells with a high CD133 expression both in vivo and in vitro.

AC133 Antigen ; Animals ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Apoptosis ; Carcinoma, Hepatocellular ; drug therapy ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cisplatin ; pharmacology ; Glycoproteins ; immunology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Peptides ; immunology ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the effect of amphotericinB (AmB) on migration and invasion of esophageal carcinoma Eca109 cells exposed to hypoxia and explore the molecular mechanisms.

METHODSRoutinely cultured esophageal carcinoma Eca109 cells were treated with 0, 1.25, 2.5, or 5 µg/ml AmB in hypoxic condition (3% O2, 5% CO2, and 92% N2) for 24 h. The cell migration and invasion were assessed by cell scratch test and Transwell chamber assay, respectively. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase-2 (MMP-2), and E-cadherin in the cells, respectively.

RESULTSCompared with the control cells, the cells treated with different doses of AmB showed attenuated ability of migration and invasion (P<0.05). AmB treatment resulted in significantly lowered mRNA and protein expressions of MMP-2 (P<0.05) and increased expressions of E-cadherin (P<0.05); the protein expression of HIF-1α decreased significantly in cells after AmB treatment (P<0.05) but its mRNA levels showed no significant changes (P>0.05).

CONCLUSIONAmB can suppress the migration and invasion of esophageal carcinoma Eca109 cells in hypoxic microenvironment possibly by regulating the expressions of HIF-1α, MMP-2 and E-cadherin.

Amphotericin B ; pharmacology ; Cadherins ; metabolism ; Cell Hypoxia ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Down-Regulation ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; RNA, Messenger

BACKGROUNDHemifacial spasm (HFS) is a facial nerve disorder characterized by episodic involuntary ipsilateral facial muscle contraction. Information on Chinese patients with HFS has not been well-characterized. This study aimed to evaluate the clinical feature and the treatment status of HFS across China.

METHODSA cross-sectional study including 1003 primary HFS patients had been carried out in 15 movement disorder clinics in China in 2012. The investigated information was acquired from questionnaires and medical records including demographic data, site of onset, aggravating and relieving factors, treatments prior to the investigation, etc.

RESULTSIn this study, the ratio of male to female was 1.0:1.8, the mean age at onset was (46.6 ± 11.5) years. About 1.0% patients were bilaterally affected. The most often site of initial onset was the orbicularis oculi muscle. The most often affected sites were orbicularis oculi, zygomatic, and orbicularis oris muscles. Stress/anxiety and relaxation were most often aggravating and relieving factors, respectively; 2.3% patients had family history, 28.4% cases were combined with hypertension, and 1.4% patients were with trigeminal neuralgia. Botulinum toxin type A (BTX-A) injection was the most commonly used treatment, followed by acupuncture and oral medication. BTX-A maintained the highest repeat treatment ratio (68.7%), while 98.4% patients gave up acupuncture. The mean latency of BTX-A effect was (5.0 ± 4.7) days, the mean total duration of the effect was (19.5 ± 11.7) weeks, and 95.9% patients developed improvements no worse than moderate in both severity and function. The most common side effect was droopy mouth.

CONCLUSIONSThe onset age of HFS in China is earlier than that in western countries. The most often used two treatments are BTX-A injection and acupuncture, while the latter kept the poor repeat treatment ratio because of dissatisfactory therapeutic effect.

Adult ; Botulinum Toxins, Type A ; therapeutic use ; China ; Cross-Sectional Studies ; Female ; Hemifacial Spasm ; diagnosis ; drug therapy ; Humans ; Male ; Middle Aged ; Neuromuscular Agents ; therapeutic use

Objective To investigate the effect of amphotericinB (AmB) on migration and invasion of esophageal carcinoma Eca109 cells exposed to hypoxia and explore the molecular mechanisms. Methods Routinely cultured esophageal carcinoma Eca109 cells were treated with 0, 1.25, 2.5, or 5μg/ml AmB in hypoxic condition (3%O2, 5%CO2, and 92%N2) for 24 h. The cell migration and invasion were assessed by cell scratch test and Transwell chamber assay, respectively. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase-2 (MMP-2), and E-cadherin in the cells, respectively. Results Compared with the control cells, the cells treated with different doses of AmB showed attenuated ability of migration and invasion (P<0.05). AmB treatment resulted in significantly lowered mRNA and protein expressions of MMP-2 (P<0.05) and increased expressions of E-cadherin (P<0.05); the protein expression of HIF-1α decreased significantly in cells after AmB treatment (P<0.05) but its mRNA levels showed no significant changes (P>0.05). Conclusion AmB can suppress the migration and invasion of esophageal carcinoma Eca109 cells in hypoxic microenvironment possibly by regulating the expressions of HIF-1α, MMP-2 and E-cadherin.

Objective To study the inhibitory effect of CD133 monoclonal antibody labeled with 131I (131I-CD133mAb) on Huh-7 human liver cancer cell line overexpressing CD133 antigen in vitro and in mouse models bearing the tumor cell xenograft. Methods 131I-CD133mAb was prepared by chloramines-T method and evaluated for its stability. Flow cytometry and immunohistochemistry were used to detect the expression of CD133 in Huh-7 cells and in Huh-7 cell-derived tumors, respectively. Huh-7 cells treated with 131I-CD133mAb plus cisplatin (DDP), 131I-CD133mAb, DDP, or no treatment (blank control) were examined for cell proliferation suppression by MTT assay with the IC50 calculated. BALB/c mice bearing subcutaneous Huh-7 cell xenograft in the right forelegs were treated with 131I-CD133mAb, DDP, or both every two days for two weeks. The tumor size and volume were measured twice a week, and pathological examination of the tumor was carried out after the treatments. The tumor inhibition rate was calculated and tumor cell apoptosis observed with HE staining. Results The labeling ratio of 131I-CD133mAb was 90.25% and the radiochemical purity was 97.78%. Huh-7 cells showed obviously higher CD133 expression than HepG2 cells. 131I-CD133mAb combined with DDP group resulted in a significantly higher tumor inhibition rate than other treatments in the tumor-bearing mice. Conclusion 131I-CD133mAb can inhibit the growth of liver cancer cells with a high CD133 expression both in vivo and in vitro.

Objective To investigate the effect of amphotericinB (AmB) on migration and invasion of esophageal carcinoma Eca109 cells exposed to hypoxia and explore the molecular mechanisms. Methods Routinely cultured esophageal carcinoma Eca109 cells were treated with 0, 1.25, 2.5, or 5μg/ml AmB in hypoxic condition (3%O2, 5%CO2, and 92%N2) for 24 h. The cell migration and invasion were assessed by cell scratch test and Transwell chamber assay, respectively. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase-2 (MMP-2), and E-cadherin in the cells, respectively. Results Compared with the control cells, the cells treated with different doses of AmB showed attenuated ability of migration and invasion (P<0.05). AmB treatment resulted in significantly lowered mRNA and protein expressions of MMP-2 (P<0.05) and increased expressions of E-cadherin (P<0.05); the protein expression of HIF-1α decreased significantly in cells after AmB treatment (P<0.05) but its mRNA levels showed no significant changes (P>0.05). Conclusion AmB can suppress the migration and invasion of esophageal carcinoma Eca109 cells in hypoxic microenvironment possibly by regulating the expressions of HIF-1α, MMP-2 and E-cadherin.

Objective To study the inhibitory effect of CD133 monoclonal antibody labeled with 131I (131I-CD133mAb) on Huh-7 human liver cancer cell line overexpressing CD133 antigen in vitro and in mouse models bearing the tumor cell xenograft. Methods 131I-CD133mAb was prepared by chloramines-T method and evaluated for its stability. Flow cytometry and immunohistochemistry were used to detect the expression of CD133 in Huh-7 cells and in Huh-7 cell-derived tumors, respectively. Huh-7 cells treated with 131I-CD133mAb plus cisplatin (DDP), 131I-CD133mAb, DDP, or no treatment (blank control) were examined for cell proliferation suppression by MTT assay with the IC50 calculated. BALB/c mice bearing subcutaneous Huh-7 cell xenograft in the right forelegs were treated with 131I-CD133mAb, DDP, or both every two days for two weeks. The tumor size and volume were measured twice a week, and pathological examination of the tumor was carried out after the treatments. The tumor inhibition rate was calculated and tumor cell apoptosis observed with HE staining. Results The labeling ratio of 131I-CD133mAb was 90.25% and the radiochemical purity was 97.78%. Huh-7 cells showed obviously higher CD133 expression than HepG2 cells. 131I-CD133mAb combined with DDP group resulted in a significantly higher tumor inhibition rate than other treatments in the tumor-bearing mice. Conclusion 131I-CD133mAb can inhibit the growth of liver cancer cells with a high CD133 expression both in vivo and in vitro.

Objective To evaluate the application of CT myelography (CTM) in detecting the site of spontaneous cerebrospinal fluid (CSF) leaks and analyze it's imaging features.Methods Six patients (3 women and 3 men) with spontaneous intracranial hypotension (SIH) were included, who met the criteria of the International Headache Classification (2nd edition, 2004). Five patients subsequently underwent whole spine MRI and all 6 patients underwent CTM. Autologous blood mixed with omnipaque (300 mg/ml) was injected followed by selective puncture at the leak site indicated by CTM. Results MRI was failed to find leak site in the 5 patients, whereas CTM successfully found leak sites in all 6 patients. There were 1 to 7 leak sites respectively with an average of 4.2 sites (totally 25 points). Leak sites at cervical (12 sites) and thoracic (12 sites) were more frequent than at lumbar (1 site). CTM was featured by linear leakage of the contrast medium along the spinal nerve roots, paraspinal collections of hyper-density contrast medium and beak-like enlargement of the nerve sleeves. All patients responded well to the treatment, with complete resolution of symptoms. Conclusion CTM has been shown to be a study of choice to accurately define the location and extent of a CSF leak.

Six patients(3 male,3 female)aged 33-66 yr, Course of disease 6-35 days, diagnosed with spontaneous intracranial hypotemion in Sir Run Run Shaw Hospital from November 2007 to May 2008,Were treated with on epidural autologus blood patch trader X-ray guidance. According to the results of CT myelography. the puncture site Was chosen in the 1-2 vertebral interspaees below the site of a single eerebrospinal fluid leak. For patients with multiple cerebrospinal fluid leaks.the procedure was performed at the spinal cord segment of the most severe leak. The mixture of augous blood and Omnipaque(300g/100 ml)at a ratio of 2:1 was injected into the epidural space to cover the spinal cord segment of the leaks under C-arm fluoroscopic guidance.Side effects were recorded during and after the injection of autologus blood.The treatment was repeated one week later if the former one failed.One patient was treated with epidural blood patch 3 times.and the rest 5 patients were treated with epidural blood patch 1 time.During the injeetion of autologus blood,5 patients complained of pain in the purtclure site,2 experienced pain radiating to the upper limb,2 felt,numbness in the right leg and all of these syndromes could be self-relieved.No side effects were observed after the injection of autologus blood.All the patients were Cured and no recurrence was observed during a 2-6 month follow-up.