OBJECTIVETo explore the inhibition of Hedyotis diffusa Willd Injection (HDI) on the proliferation of RPMI 8226 cells and its mechanisms.

METHODSThe inhibition of HDI on the proliferation of RPMI 8226 cells was detected by MTT and the drug concentrations for further researches were screened out. The apoptosis rate was detected using Annexin V-PI of flow cytometry. The cell cycle distribution was detected by PI. The expressions of adhesion molecule FITC-CD44 and PE-CD49d were detected. The IL-6 and VEGF concentrations of cell supernatants were tested by ELISA. The mRNA expressions of Bax, Bcl-2, Caspase-3, Survivin, IL-6, and VEGF were detected by RT-PCR.

RESULTSHDI could inhibit the proliferation of RPMI 8226 cells. Meanwhile, it induced their early apoptosis, arresting them at G1 phase in a concentration-dependent manner. The VEGF concentrations were down-regulated after acted by 0, 20, 40, and 60 microL/mL HDI in a dose-dependent manner (P< 0.01). The IL-6 content increased (P<0.01). The expressions of CD44 and CD49d were up-regulated in a concentration-dependent manner. After acted by 40 microL/mL HDI, the Survivin mRNA level was significantly downregulated (P<0.01), the mRNA levels of Bcl-2, IL-6, and VEGF were significantly up-regulated (P<0.01), but the up-regulation of Bax and Caspase-3 mRNA levels were not so obvious (P>0.05).

CONCLUSIONSHDI could inhibit the proliferation of RPMI 8226 cells. Its mechanisms might be correlated with early apoptosis induction, G1 phase arresting, VEGF secretion lowering, and Survivin mRNA transcription level down-regulating.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Hedyotis ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Interleukin-6 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia.

METHODSWe retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage.

RESULTSGroup A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05).

CONCLUSIONThe rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

Azoospermia ; therapy ; Epididymis ; cytology ; physiopathology ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Spermatozoa ; Testis ; cytology ; physiopathology

OBJECTIVETo identify protein markers for the early diagnosis of pancreatic cancer by a comparative proteomic method.

METHODSComparative analysis on the pancreatic peripheral blood protein profiling from 20 pancreatic cancer patients, 10 chronic pancreatitis patients and 20 cancer-free controls from May 2007 to September 2008 was carried out by two-dimensional fluorescence electrophoresis (2D-DIGE). Differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The significance difference proteins were confirmed by Western-blot.

RESULTSA differentially expressed proteins: complement 3 (C3) was identified. The gray level of C3 in pancreatic cancer tissue, chronic pancreatitis, and normal control group were 1.63 ± 0.28, 0.65 ± 0.13 (t = 11.81, P = 0.00) and 0.88 ± 0.19 (t = 9.93, P = 0.00), respectively. C3 was high expression in pancreatic cancer group compared with normal control group. The expression of C3 was higher in pancreatic cancer group than in chronic pancreatitis group. The high expression of C3 in pancreatic carcinoma was confirmed by Western blot.

CONCLUSIONS2D-DIGE and MALDI-TOF-MS technology is a quick, easy and practical method to screen for specific biomarkers in serum of patients with pancreatic carcinoma. The identified protein C3 in this study may be as specific serum biomarkers of pancreatic carcinoma.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Case-Control Studies ; Complement C3 ; analysis ; Early Diagnosis ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; diagnosis ; Pancreatitis, Chronic ; blood ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Two-Dimensional Difference Gel Electrophoresis

OBJECTIVETo compare therapeutic effects between Kirschner wire fixation after early debridement (Kirschner wire group) and staged plate fixation (plate group) in the treatment of open calcaneal fractures.

METHODSFrom January 2001 to May 2008, 55 patients (58 feet) with open calcaneal fractures were reviewed,the mean age was 36.8 years(ranged, 19 to 65 years) and the average visit time was 3 hours (ranged, 30 min to 7 h). All the patients were divided into two groups: Kirschner wire group and plate group. There were 20 males (20 feet) and 9 females (9 feet) in Kirschner wire group, in which 15 feet were type I, 13 feet were type II, 1 foot was type III A according to Gustilo classification and 9 feet were type II, 18 feet were type III, 2 feet were type IV according to Sanders classification. The patients in Kirschner wire group were treated with early debridement, fracture reduction and Kirschner wire fixation, and the soft tissue defects were covered with VSD temporarily, and then were enveloped by skin or flap grafts at the second stage. There were 18 males (19 feet) and 8 females (10 feet) in the plate group, in which 13 feet were type I, 14 feet were type II, 2 feet were type III A according to Gustilo classification and 11 feet were type II,15 feet were type III, 3 feet were type IV according to Sanders classification. The patients in the plate group were treated with early debridement, and plate internal fixation with were performed when the wound became stabilization.

RESULTSTwenty-three feet (15 patients) in the Kirschner wire group and 22 feet (13 patients) in the plate group were followed, the duration ranged from 10 to 36 months, with an average of 24 months. Compared with preoperative ones, the heel height, width, Böhler angle and Gissane angle of calcaneal got improvements. According to AOFAS ankle-foot evaluation system, 11 feet got an excellent result, 8 good in the Kirschner wire group; 2 feet had wound local skin necrosis and cured by dressing; 1 foot had a large area of skin necrosis and deep infection; 1 foot had chronic osteomyelitis. All above 4 feet underwent arthrodesis later. As comparison, 7 feet got an excellent result, 4 good in the plate group; 2 patients had mild complications of wounds; 1 patient had chronic osteomyelitis after early debridement; 10 patients had wound complications after internal fixation, including 7 patients with skin necrosis, superficial infection in 3 patients. There were statistical significant in radiological indicators and AOFAS ankle-foot scores between two groups. But there were no significant differences in early postoperative complications between the two groups.

CONCLUSIONEarly debridement and Kirschner wire fixation for the treatment of open calcaneal fractures has fewer early complications, which is a simple, safe and effective method.

Adult ; Aged ; Bone Plates ; Bone Wires ; Calcaneus ; injuries ; surgery ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; instrumentation ; Fractures, Bone ; surgery ; Fractures, Open ; surgery ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To construct natural immune Fab fragment phage display library and to screen the antibodies against human colorectal cancer. Methods Total RNA in the metastatic lymph nodes from patients with colorectal cancer was extracted routinely, and reverse transcriptase PCR employed to amplify the heavy chain Fd and light chain κ cDNA. The amplification products were inserted in succession into the vector pComb3 to construct human Fab fragment library, which was screened and identified using phage display technique. Results The heavy chain Fd fragment and light chain κ fragment (650 bp in length) were amplified, and the products were inserted into pComb3 vector with a recombination rate of 40%. The constructed phage display Fab library had a capacity of up to 1.48×106, and was enriched approximately 120 times after 3 cycles of panning. Dot immunoblotting found the expression of Fab fragment which could bind to the antigens of human colorectal cancer. Conclusion The phage display library of the antibody Fab fragment from naturally immunized lymph nodes is successfully established, rendering it possible to screen the antibodies against human colorectal cancer.

Objective To construct natural immune Fab fragment phage display library and to screen the antibodies against human colorectal cancer. Methods Total RNA in the metastatic lymph nodes from patients with colorectal cancer was extracted routinely, and reverse transcriptase PCR employed to amplify the heavy chain Fd and light chain κ cDNA. The amplification products were inserted in succession into the vector pComb3 to construct human Fab fragment library, which was screened and identified using phage display technique. Results The heavy chain Fd fragment and light chain κ fragment (650 bp in length) were amplified, and the products were inserted into pComb3 vector with a recombination rate of 40%. The constructed phage display Fab library had a capacity of up to 1.48×106, and was enriched approximately 120 times after 3 cycles of panning. Dot immunoblotting found the expression of Fab fragment which could bind to the antigens of human colorectal cancer. Conclusion The phage display library of the antibody Fab fragment from naturally immunized lymph nodes is successfully established, rendering it possible to screen the antibodies against human colorectal cancer.

OBJECTIVEWe observed the therapeutic effectiveness and safety of different antidepressants as well as the correlation between symptomatic improvement of depression and improvement of chest pain in patients with susceptible "angina pectoris" and negative coronary angiogram complicating comorbid depression.

METHODSIn this double-blinded randomized study, a total of 123 eligible patients were allocated into three groups: (1) Group F: fluoxetine 20 mg QN (n = 41); (2) Group P: Placebo 1 tablet QN (n = 40); (3) Group F + O: fluoxetine 20 mg + olanzapine 2.5 mg QN for the former 2 weeks and only fluoxetine 20 mg QN for the latter 2 weeks (n = 42). The total therapy duration was 4 weeks. HAMD, HAMA and self-evaluation table of chest pain were obtained before therapy, at the end of 1 and 2 weeks after therapy.

RESULTSBaseline HAMD and HAMA scores and self-evaluation score of chest pain were similar among 3 groups and all scores were significantly improved post various therapies in the order of group F + O > group F > group P. The rate of score decrease were seen after 1 week treatment in group F + O and after 2 week treatment in group F. There was a significant positive correlation between the rates of self-evaluation chest pain score decrease and HAMD (r = 0.867, P < 0.001) and HAMA (r = 0.854, P < 0.001) score decreases after 4 weeks therapies (P < 0.05). During the whole course of treatment, no serious adverse reaction was found in all patients.

CONCLUSIONIn patients with suspected "angina pectoris" and negative coronary angiogram complicating comorbid depression, the antidepressants were safe and significantly improved the symptoms of depression and anxiety and chest pain. Low dose fluoxetine plus short term olanzapine regimen was superior to fluoxetine alone regimen in terms of stronger and quicker symptom improvement.

Aged ; Angina Pectoris ; diagnostic imaging ; drug therapy ; psychology ; Antidepressive Agents, Second-Generation ; therapeutic use ; Benzodiazepines ; therapeutic use ; Coronary Angiography ; Depressive Disorder ; drug therapy ; etiology ; Double-Blind Method ; Female ; Fluoxetine ; therapeutic use ; Humans ; Male ; Middle Aged

OBJECTIVES: To establish a gas chromatography-mass spectrometry （GC-MS） method for the determination of sulfide ion in blood and apply it to the practical cases. METHODS: The 1, 3, 5-tribromobenzene was selected as an internal standard, and 0.2 mL blood sample was collected and analyzed using GC-MS after α-Bromo-2, 3, 4, 5, 6-pentafluorobenzyl bromide derivatization. RESULTS: The mass concentration of sulfide ion in blood had good linearity in the range of 0.2-40 μg/mL with a limit of detection （LOD） of 0.05 μg/mL. The mass concentration of sulfide ion was less than 0.05 μg/mL in blank blood from different sources such as healthy subjects and dead cases. In 3 sulfide poisoning cases, sulfide ion was detected in the blood samples of 6 victims, and the mass concentration range was 1.02-3.13 μg/mL. CONCLUSIONS This study establishes a method for investigation of sulfide ion in blood which has been applied successfully to the cases of fatal sulfide poisonings.
Fluorobenzenes ; Gas Chromatography-Mass Spectrometry/methods* ; Humans ; Hydrogen Sulfide/blood* ; Limit of Detection ; Sulfides

Exosomes are involved in invasion, migration and angiogenesis of tumor cells, and invasion is the main cause of death in glioma patients. Studies have shown that the exosomes secreted by tumor cells can carry miRNA into the receptor cells and regulate the biological functions of the receptor cells, such as proliferation, migration and invasion. miR-574-5p plays a key role in the occurrence and development of a variety of tumors. However, whether the exosomes derived from glioma cells express miR-574-5p and its role in the growth, invasion and migration of glioma cells have not been reported. This study investigated the mechanism of the exosomal miR-574-5p secreted from glioma cells in the process of cell proliferation, migration and invasion. The exosomes were characterized by electron microscopy, nanoparticle size tracking and Western blot. The results displayed that the extracted exosomes were round particles with a diameter of 30 ~ 100 nm. The internalization of exosomes was detected by immunofluorescence assay. The results showed that exosomes were internalized into LN229 cells; Bioinformatics and online data were used to screen the differentially secreted miRNA between LN229 and H4 glioma cells. The results showed that the differentially secreted miRNA was miR-574-5p, and large tumor suppressor 2 (LATS2) was predicted to be the target gene of miR-574-5p; Duel luciferase reporter assay confirmed that miR-574-5p was complementary to the 3'UTR region of LATS2; The transfection assay, qRT-PCR and Western blot was conducted to measure the relationship between miR-574-5p and LATS2. The results demonstrated that there was no significant difference in LATS2 mRNA levels between the control group and the group with miR-574-5P overexpression (P > 0.05), suggesting the regulatory effect of miR-574-5P on LATS2 was achieved by inhibiting its translation (P < 0.05). CCK-8, Transwell migration and invasion assays were conducted to explore the effect of miR-574-5p on proliferation, migration and invasion of LN229 cells. The results showed that overexpression of miR-574-5p could significantly promote the ability of proliferation, migration and invasion of LN229 cells (P < 0.05). In addition, Western blot was performed to measure the expression of kinase proteins involved in the LATS2/YAP signaling pathway, and the influence of the exosomes on this signaling pathway. The results revealed that the exosomes down-regulated the protein expression level of LATS2 and reduced p-YAP phosphorylation. In conclusion, the exosomal miR-574-5p can promote the proliferation, migration and invasion of glioma cells by down-regulating LATS2 and activating LATS2/YAP signaling pathway, which may provide a potential biomarker for the diagnosis and target for the treatment of glioma.

Neural stem cell therapy, as a new therapeutic method for neural diseases, has aroused a wide concern for over 20 years since neural stem cells were first found in 1992. Ischemic stroke is highly concerned because of its high incidence, mortality and disability rates. Because the brain has a limited ability to repair itself, to improve neural function and promote neural regeneration may help to prevent occurrence and development of neurological diseases. It is noteworthy that some stroke patients showed an ability to repair brain several months after the stroke happened, suggesting an existence of endogenous nerve repair in these patients. The research advances in functions of endogenous neural stem cells in neural regeneration and the related regulators after ischemic stroke are summarized in this review to provide new views of the mechanism of neural functional recovery after ischemic stroke.
Brain Ischemia ; therapy ; Humans ; Nerve Regeneration ; Neural Stem Cells ; cytology ; Stroke ; therapy