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

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

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.

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

OBJECTIVE: To compare the therapeutic effect of electro-nape-acupuncture (ENA) combined with hyperbaric oxygen therapy (HBOT) and single HBOT on refractory flat descending idiopathic sudden sensorineural hearing loss (ISSNHL). METHODS: A total of 78 patients were randomized into an ENA combined with HBOT (ENA+HBOT) group and a HBOT group, 39 cases in each one. Patients in both groups were treated with oral extract of ginkgo biloba leaves and mecobalamin tablets. On the basis of the conventional medication treatment, HBOT was adopt in the HBOT group. On the basis of the treatment in the HBOT group, electro-nape-acupuncture was applied at Fengchi (GB 20), Gongxue (Extra), Zhongzhu (TE 3), Waiguan (TE 5) and Yifeng (TE 17), Tinggong (SI 19), Tinghui (GB 2) and the vertigo-auditory area of affected side in the ENA+HBOT group. Pulse acupuncture instrument was connected at Fengchi (GB 20) and Gongxue (Extra) for 30 min (with continuous wave, 2 Hz in frequency), the needles were retained for another 30 min after electroaupuncture. The treatment was given once a day, 6 times a week for 4 weeks in both groups. Before the treatment and 2,4 weeks into the treatment, the average auditory threshold, the scores of tinnitus handicap inventory (THI) and dizziness handicap inventory (DHI) were observed, and the therapeutic effect was evaluated in both groups. RESULTS: Compared before treatment, the average auditory threshold, the scores of THI and DHI of 2,4 weeks into the treatment were decreased in both groups (<0.000 1). Compared with the HBOT group, the average auditory threshold, the scores of THI and DHI of 4 weeks into the treatment were lower in the ENA+HBOT group (<0.000 1). The total effective rate was 69.2% (27/39) in the ENA+HBOT group and 51.3% (20/39) in the HBOT group, there was no statistical difference (>0.05). CONCLUSION Electro-nape- acupuncture can improve the mean auditory threshold and the symptoms of tinnitus and dizziness in patients with refractory flat descending idiopathic sudden sensorineural hearing loss.
Acupuncture Therapy ; Dizziness ; therapy ; Hearing Loss, Sensorineural ; therapy ; Hearing Loss, Sudden ; therapy ; Humans ; Hyperbaric Oxygenation ; Plant Extracts ; therapeutic use ; Tinnitus ; therapy ; Treatment Outcome ; Vitamin B 12 ; analogs & derivatives ; therapeutic use

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

OBJECTIVE: To evaluate the associations of lipid indicators and mortality in Beijing Elderly Comprehensive Health Cohort Study. METHODS: A prospective cohort was conducted based on Beijing Elderly Comprehensive Health Cohort Study with 4499 community older adults. After the baseline survey, the last follow-up was March 31, 2021 with an average 8.13 years of follow-up. Cox proportional hazard model was used to estimate the hazard ratios (HR) with 95% CI for cardiovascular disease (CVD) death and all-cause death in associations with baseline lipid indicators. RESULTS: A total of 4499 participants were recruited, and the mean levels of uric acid, body mass index, systolic blood pressure, diastolic blood pressure, fasting plasma glucose, total cholesterol (TC), triglyceride, and low-density lipoprotein cholesterol (LDL-C) showed an upward trend with the increasing remnant cholesterol (RC) quarters (P_trend < 0.05), while the downward trend was found in high-density lipoprotein cholesterol (HDL-C). During the total 36,596 person-years follow-up, the CVD mortality and all-cause mortality during an average 8.13 years of follow-up was 3.87% (95% CI: 3.30%-4.43%) and 14.83% (95% CI: 13.79%-15.86%) with 174 CVD death participants and 667 all-cause death participants. After adjusting for confounders, the higher level of TC (HR = 0.854, 95% CI: 0.730-0.997), LDL-C (HR = 0.817, 95% CI: 0.680-0.982) and HDL-C (HR = 0.443, 95% CI: 0.271-0.724) were associated with lower risk of CVD death, and the higher level of HDL-C (HR = 0.637, 95% CI: 0.501-0.810) were associated with lower risk of all-cause death. The higher level of RC (HR = 1.276, 95% CI: 1.010-1.613) increase the risk of CVD death. Compared with the normal lipid group, TC ≥ 6.20 mmol/L group and LDL-C ≥ 4.10 mmol/L group were no longer associated with lower risk of CVD death, while RC ≥ 0.80 mmol/L group was still associated with higher risk of CVD death. In normal lipid group, the higher levels of TC, LDL-C and HDL-C were related with lower CVD death. CONCLUSIONS In community older adults, higher levels of TC and HDL-C were associated with lower CVD mortality in normal lipid reference range. Higher RC was associated with higher CVD mortality, which may be a better lipid indicator for estimating the CVD death risk in older adults.