Objective To evaluate the significance of laparoscopy in the diagnosis and treatment of unexplained acute right lower abdominal pain in women. Methods Clinical data of 52 women with unexplained acute right lower abdominal pain from April 1999 to April 2004 were retrospectively analyzed. Results Diagnosis was clarified under laparoscope in all the 52 patients, including 28 cases of acute appendicitis, 18 cases of acute pelvic inflammatory disease, 3 cases of ovarian cyst pediculotorsion, 2 cases of right tubal pregnancy, and 1 case of Crohn’s disease with iliac perforation. With exception of a conversion to open surgery in 3 patients, laparoscopic treatment was successfully completed in the remaining 49 patients. No surgical complications occurred. The patients were discharged from the hospital 2~6 days postoperatively. Conclusions Laparoscopy is a timely, safe and effective alternative for the diagnosis and treatment of unexplained acute lower right abdominal pain in women.

Objective To observe the effect of Dachengqi decoction combined with β-aescine on pulmonary function in patients with acute lung injury (ALI) after chest trauma.Methods Ninety-five patients with ALI after chest trauma admitted to the Department of Thoracic Cardiothoracic Surgery of Dingxi People's Hospital of Gansu from April 2013 to May 2016 were enrolled, and they were divided into a Dachengqi decoction group (47 patients) and a control group (48 patients) by random number table. The control group patients were treated with conventional therapy and β-aescine 0.4 mg/kg+ 10% Glucose (250 mL) intravenous drip, while the Dachengqi decoction group patients were additionally treated with Dachengqi decoction on the treatment of the control group (rhubarb 12 g, magnolia officinalis 24 g, fructus aurantii immaturus 12 g, mirabilite 9 g), one dose daily, taken twice in a day, once in the morning and once in the evening orally, both groups were treated for one week. Before and after treatment, the changes of esophageal pressure, respiratory dynamics, the indexes of pulmonary function and blood gas analysis were observed. Results Compared with those before treatment, the levels of plateau pressure (Pplat), mean airway pressure (mPaw), airway resistance (Raw) and arterial partial pressure of carbon dioxide (PaCO2) of the two groups after treatment were significantly reduced, but the levels of lower esophageal sphincter pressure (LESP), peak airway pressure (PIP), the first second forced expiratory volume (FEV1), forced vital capacity (FVC), peak expiratory flow rate (PEF), the mean maximum expiratory flow (MMEF), pulse oxygen saturation (SpO2), arterial partial pressure of oxygen (PaO2), oxygenation index (PaO2/FiO2) were all increased significantly; the changes of the above indexes in the Dachengqi decoction group were more significant than those in the control group [LESP (mmHg, 1 mmHg = 0.133 kPa):18.64±2.79 vs. 15.46±3.09, Pplat (cmH2O, 1 cmH2O = 0.098 kPa): 14.27±1.68 vs. 16.00±1.87, PIP (cmH2O): 40.23±5.03 vs. 32.19±4.45, mPaw (cmH2O): 8.57±0.67 vs. 9.41±1.23, Raw (cmH2O·L-1·s-1): 6.76±1.01 vs. 9.31±1.43, FEV1 (L): 1.73±0.27 vs. 1.46±0.25, FVC: (3.95±0.51)% vs. (3.30±0.46)%, PEF (L/s): 3.81±0.47 vs. 3.11±0.38, MMEF (L/s): 0.93±0.16 vs. 0.77±0.12, SpO2: 0.96±0.06 vs. 0.91±0.05, PaO2 (mmHg): 97.82±10.13 vs. 74.73±8.02, PaCO2 (mmHg): 36.49±4.28 vs. 47.13±5.46, PaO2/FiO2 (mmHg): 362.47±20.82 vs. 259.79±17.61, allP < 0.05]. Conclusion Dachengqi decoction combined with β-aescine can reduce the respiratory intensity of patients with ALI after chest trauma and improve the lung function and blood oxygen level.

Objective To observe cell proliferation and neurogenesis in the pregnant mouse dentate gyrus (DG) and the subventricular zone(SVZ). Methods Injection of the thymidine analog 5 bromo 2 deoxyuridine (BrdU)to determine the extent of cell proliferation combined with single or double immunohistochemistry staining with antibodies BrdU,TuJ1(class Ⅲ ? tubulin,neuron specific early differentiation marker)and GFAP. Results The number of BrdU positive cells in the pregnant mouse dentate gyrus was significantly more than that of unpregnant mouse dendate gyrus but not in the subventricular zone.In dentate gyrus,approximately 80% of these cells were neuronal characteristics (TuJ1 immunoreactive)and 3%～5% of these cells were astrocytic characteristics(GFAP immunoreactive).Conclusion\ These findings suggest that pregnancy significantly increases cell proliferation and neurogenesis in the adult mouse dentate gyrus and present the possibility that these new cells exert an important influence on hippocampal function

Objective To isolate neuroepithelial stem cells from the spinal cord neural tube of the embryonic rat and induce them to differentiate into dopaminergic neurons. Methods Serum free cells suspension culture and single cell cloning technique were used to isolate neuroepithelial stem cells. 5 bromo 2 deoxyuridine(BrdU) to label new cells combined with single or double immunocytochemistry staining to detect nestin antigen before differentiation and neural cell specific antigens after differentiation, such as neurofilament (NFM 160?kD), glial fibrillary acidic protein(GFAP), galactocerebroside(GalC) and tyrosine hydroxylase(TH). Striatal extracts were used to induce neuroepithelial stem cells to differentiate into dopaminergic neurons. Results The cells isolated from the spinal cord neural tube of the embryonic rat expressed nestin antigen. They had the potential to serially passage and differentiate into neurons, astrocytes and oligodendrocytes. Striatal extracts could induce 12% of them to differentiate into dopaminergic neurons compared with 3% in controls.Conclusion The cells, which express nestin antigen, isolated from neural tube are multipotent and have the ability to self renew, therefore, they are neural stem cells. These stem cells can be induced to differentiate into specific neurons in vitro. Which can provide materials for neural transplantation.

BACKGROUND: The skill to culture osteoblasts primarily has been well developed. However, trypsinase can affect membrane protein of osteoblasts if the time of digestion is long. Therefore, it is of great significance to select an ideal method to avoid the damage from trypsinase to cells as possible when culturing osteoblasts.OBJECTIVE: To explore a novel method to isolate and culture SD rat osteoblasts in vitro, and identify the functions of the cells.DESIGN: Observational study.SETTING: Department of Orthopaedics, First Affiliated Hospital of Jinan University.MATERIALS: This experiment was carried out in the Department of Orthopaedics, First Affiliated Hospital of Jinan University from March to May in 2007. Eight SPF 24-hour old SD rats were used in the experiment. The rats, irrespective of gender, were provided by the Experimental Animal Center of Nanfang Medical University. The experimental animals were disposed according to ethical criteria. The main reagents were detailed as follows: collagenase Ⅱ (Sigma Company);trypsin (Sigma Company); alkaline phosphatase (ALP) kit (Nanjing Jiancheng Biological Products Company); SABC-1021(Wuhan Boster Biotechnology Company).METHODS: 24-hour old SD rats were chosen for experiment. The newly born SD rats were sacrificed by anesthesia and the cranial bones of the rats were obtained cleanly, erased completely of the periosteum and cut to blocks of I mm3. The cranial bones were digested by 0.25 % trypsinase for 20 minutes, then by 0. 1% type Ⅱ collagenase for 60 minutes. The digestive time of trypsinase was controlled in the process of digestion to avoid to harm the cells. The liquid was gathered and centrifuged. The cells were cultured in culture flask and were purified by many times adhered.MAIN OUTCOME MEASURES: Morphology observations under the inverted phase contrast microscope, transmission electron microscope, and scanning electron microscope were performed. The phenotype, calcium tuberculation and the expression of alkaline phosphatase were studied with alizarin red staining and modified Gomori Ca-Co assays respectively.The cells were also evaluated with collage Ⅰ immunohistochemical staining.RESULTS: The cultured cells had active proliferation ability. Cells showed multi-angle or fusiform shape. Nucleus was immature and organell was plentiful. Therefore, they had typical morphological characters of osteoblasts. Moreover, they showed the osteoblastic phenotypes such as their synthesis of alkaline phosphatase, collage Ⅰ and formation of calcium tuberculations.CONCLUSION: The cells cultured by our modified enzymatic digestion method had typical morphological and biological characteristics of osteoblasts.

Objective To investigate the effects of fasudil hydrochloride on learning and memory, and the autophagy in hippocampal CA1 neurons in subarachnoid hemorrhage (SAH) rats. Methods Fifty-four male Sprague-Dawley rats were randomly divided into sham group (n=18), SAH group (n=18) and drug group (n=18). Subarachnoid hemorrhage model was established with internal carotid artery punc-ture. The drug group was injected fasudil hydrochloride 10 mg/kg intraperitoneally after modeling per 24 hours, while the sham group and SAH group were injected the same volume of saline. They were tested with shuttle box test 6, 24 and 72 hours after intervention, then the hippocampal CA1 area was stained with HE and immunohistochemistry to observe the morphology of cells and the expression of Beclin-1 and microtubule-associated protein 1 light chain 3 II (LC3-II). Results Compared with the sham group, the frequence of avoidance de-creased in SAH group at each time point (P<0.05), while the avoidance reaction time increased (P<0.05);the survival of neurons in hippo-campal CA1 area decreased (P<0.05), and the expression of Beclin-1 and LC3-II increased (P<0.05). Compared with SAH group, the fre-quence of avoidance increased in the drug group at each time point (P<0.05), while the avoidance reaction time decreased (P<0.05);the sur-vival of neurons in hippocampal CA1 area increased (P<0.05) and the expression of Beclin-1 and LC3-II increased further (P<0.05). Con-clusion Fasudil hydrochloride can improve learning and memory ability and protect neurons from damage, which may associate with the ex-cess of autophagy activation in hippocampal CA1 areas in SAH rats.

[ABSTRACT]AIM:Toexploretheeffectoftheelasticmodulusandsizesofliquidcrystal(LC)phasesonosteo-genic differentiation based on OPC/PU composite substrate by mimicking the microenvironment in rat bone mesenchymal stem cells (rBMSCs).METHODS: A series of composite substrates with different elastic modulus were constructed via modulation of LC content in the composites .The surface phase structure was observed by polarized microscopy , and the mechanical property was measured by a universal material testing machine .Furthermore, the laser confocal microscope was employed to observe the spreading , polarization and the cytoskeleton arrangement of the rBMSCs .The proliferation of rBM-SCs was evaluated by CCK-8 assay.The specific mRNA expression of osteogenic differentiation such as collagen Ⅰ, and osteopontin on the composite membranes was detected by real-time PCR.RESULTS:The size and number of LC phase in-creased and the elastic modulus of the composite substrates decreased with the increase of the LC content .The rBMSCs ex-hibited better characteristics of initial adhesion , spreading and proliferation on the OPC 10-PU and OPC30-PU in the early and medium culturing .The rBMSCs displayed higher expression of collagen Ⅰ and osteopontin on the OPC10-PU in the early and medium osteogenic induction , while the high expression of these osteogenic genes occured on the OPC 30-PU and OPC50-PU in later osteogenic induction .The emphasis of genetic expression was switched from collagen Ⅰin the early and medium osteogenic induction to osteopontin in the later stage .CONCLUSION:When the content of LC remained low in the composite substrates , rBMSCs mainly responded to the mechanical stimuli induced by substrate stiffness and exhibited distinguished cellular behaviors;with the increase in the LC content , rBMSCs had strong interactions with LC by sensing the viscoelasticity of LC , probably resulted from the contribution of both substrate stiffness and the viscoelasticity of LC phase .

BACKGROUND:Correlation between subchondral bone and articular cartilage in the process of osteoarthritis has not been fuly elucidated. Degeneration of cartilage is the focus of attention, and the subchondral bone also plays an important role in the process of osteoarthritis. OBJECTIVE: To observe the differences between experimental osteoarthritis models in rabbit knees established by two kinds of surgical methods and two kinds of proteases inducing methods, and to explore the correlation between subchondral bone mass and degeneration of cartilage. METHODS:Thirty-two New Zealand rabbits were randomly and averagely divided into four groups: Hulth group (group A), anterior cruciate ligament transaction group (group B), colagenase type II group (group C) and papain group (group D). The right knees of rabbits were established as osteoarthritis models, and the left knees served as controls. Bone mineral density of the knee joint was evaluated by dual-energy X-ray absorptiometry scanning at 0, 4 and 8 weeks after modeling. The rabbits were sacrificed at 8 weeks after MRI scanning, bilateral knee joints were harvested for general and histological observation. Quantitative analysis was done according to Mankin scores. RESULTS AND CONCLUSION: Bone mineral density of the right knees decreased at 0, 4 and 8 weeks after modeling, and the rank was as folows: group A > group B > group C > group D. MRI scanning showed that the articular cartilage thickness of the medial and lateral femoral condyle on the right knees became thinner compared with the left side, and the rank was as folows: group A < group B < group C < group D. Observation by specimens and pathological slices showed that the articular cartilage degeneration of the surgery groups worsened, group A was the most serious one, and group 1D was the lightest. Both surgery and proteases inducing methods can successfuly establish osteoarthritis models in rabbit knees. Surgery inducing models resemble the advanced or intermediate stage of osteoarthritis, while the proteases inducing models resemble the early stage of osteoarthritis. Degeneration of the articular cartilage and changes of subchondral bone are related in progressive development.

Objective To observe the changes of expressions of brain-derived neurotrophic factor (BDNF)and its receptor tyrosine kinase B(TrkB)and in rats with hemitransectional spinal cord injury(SCI)after electroacupuncture on Du Meridian and rehabilitation training.Methods The animal model of acute hemitransectional lesion at the right half of T11 spinal cord was established in 96 adult female rats,which were then divided randomly into an electroacupuncture group,a rehabilitation training group,an electroacupuncture combined with rehabilitation training group and a control group.All the groups received treatment on the 3rd d after operation.The electroacupuncture group and rehabilitation training group were given electroacupuncture on points of Du Meridian and rehabilitation training,respectively,and the combined group was given Du Meridian electroacupuncture in addition to rehabilitation training.Basso-Beattie-Bresnahan(BBB)scale was used to evaluate motor function every week.Twelve rats of each group were sacrificed 4 and 8 weeks after operation,respectively,and their spinal cord tissues were extracted.The polymerase chain reaction(PCR),reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemical techniques were used to detect the expressions of BDNF and TrkB.Results BBB grade increased gradually as time went on.There were significant differences between control group and other groups at the same time point(P ＜ 0.05).The scores increased obviously in electroacupuncture combined with rehabilitation training group compared with electroacupuncture group and rehabilitation training group(P ＜ 0.05).The result of immunohistochemical observation and RT-PCR also showed that there were significant differences of expressions of BDNF and TrkB among control group and other groups at the same time(P ＜ 0.05).The effects in electroacupuncture combined with rehabilitation training group were much more obvious than those in electroacupuncture group and rehabilitation training group(P ＜ 0.05),but there was no significant difference between electroacupuncture group and rehabilitation training group (P ＞ 0.05).Conclusions Electroacupuncture on Du Meridian combined with rehabilitation training had synergic effect on rat's SCI,and could obviously improve the restoration of rat's motor function; the mechanism maybe related to the upregulation of expressions of BDNF and TrkB.

Child ; Female ; Head and Neck Neoplasms ; Humans ; Multiple Endocrine Neoplasia Type 2b ; Proto-Oncogene Proteins c-ret ; genetics