Volume 86  ·   Issue 39  ·  2006

Objective to explore a modified surgical method that can make up for the ethical defects of the classical bipedal rat model.Methods Twenty SD rats were randomly divided into modified group and traditional group.The modified group underwent brachial plexus dissection and the traditional group underwent proximal shoulder joint amputation.The active movement of shoulder joint was observed before and after the modified surgery, and the electromyography of muscles around shoulder joint was recorded to determine the extent of muscle paralysis.The surgical duration and the probability of axillary artery injury were compared with the modified method and the traditional modeling method to evaluate the surgical risk.The average daily cumulative standing time and the difference of standing posture of bipedal rats at 2 days, 1, 2, 4 and 6 weeks after surgery and 6 weeks after surgery were recorded to evaluate the effect of the modified bipedal rat animal model to simulate human standing posture.Results Compared with the traditional amputation, the modified method was simpler in operation, the operation time was shorter, and the probability of axillary vascular injury was significantly reduced. The postoperative electromyography of the modified method showed that all upper limb muscles and most of the muscles around shoulder joint were denervated, and the modified bipedal rat shoulder joint only left slight abduction motor function, After 2 weeks of operation, the average daily cumulative standing time of the modified bipedal rats was even longer than that of the classical bipedal rats, but the upright posture was no different from that of the classical bipedal rats.Conclusion The modified modeling surgical method of the bipedal rats not only simplifies the operation, but also achieves better modeling effect than the classical amputation, and completely dispels the ethical haze of the bipedal rat model.

Objective to observe the therapeutic effect of recombinant human erythropoietin (rHu-EPO) on myocardial infarction in rats and to explore its mechanism.Methods SD rat cardiomyocytes were cultured in vitro within 3 days of birth.Apoptosis was induced by hypoxia culture and H2O2 simulated oxidative stress response.Rhu-EPO was added to the culture system, and the changes of apoptosis rate were observed by fluorescence staining.32 adult SD rats were divided into blank group (8), myocardial infarction group (12) and treatment group (12); The myocardial infarction model was prepared by thoracotomy but no coronary artery ligation in the blank group, while the left anterior descending coronary artery was ligated in the myocardial infarction group and the treatment group. In the treatment group, 5000 IU/kg rHu-EPO was injected intraperitoneally every day for 7 days. After 14 days, all rats were tested for hemodynamic indexes, apoptosis was detected by in situ end labeling (TUNEL) in cardiac sections, and the expression of Bcl-2 and Bax was detected by immunohistochemistry. The rate of apoptosis induced by oxidative stress decreased from 60.42% ± 5.54% to 43.40% ± 5.39%; Intraperitoneal injection of rHu-EPO made the cardiac function of rats after myocardial infarction better preserved, and arterial systolic blood pressure, mean arterial pressure, left ventricular systolic blood pressure and ± dp/dtmax all increased, while left ventricular diastolic blood pressure and left ventricular end-diastolic pressure decreased. The apoptosis rate decreased from 1.65% ± 0.50% to 0.84% ± 0.30%; The expression rate of Bcl-2 protein increased from 0.96%±0.37% to 1.43%±0.28%; The expression rate of Bax protein decreased from 1.34%±0.47% to 0.68%±0.30%. Conclusion rHu-EPO can inhibit cardiomyocyte apoptosis and preserve cardiac function, which may be achieved by down-regulating Bax expression and up-regulating bcl-2 expression.

Objective to investigate the inhibitory effect of β-sitosterol on the growth of cervical cancer cells SiHa and its effect on the intracellular microtubule system. Methods The inhibitory effect of β-sitosterol on the proliferation of SiHa cells was determined by tetramethylazolazole blue (MTT) assay, and the changes of cell cycle before and after β-sitosterol treatment were analyzed by flow cytometry. The expression and distribution of microtubules and microtubule-associated protein 2 in SiHa cells treated with β-sitosterol were observed by laser confocal microscopy. The expression of tubulin and microtubule-associated protein 2, as well as the changes of polymerized and unpolymerized tubulin contents were detected by Western blotting. Results 20 μ mol/L β-sitosterol significantly inhibited the proliferation of SiHa cells and significantly promoted the cell aggregation in S phase. The laser confocal analysis showed that the microtubule network of SiHa cells treated with β-sitosterol for 5d was abnormal, and the expression of microtubule-associated protein 2 was significantly down-regulated. The results of Western blotting further confirmed that β-sitosterol could inhibit the expression of tubulin α and microtubule-associated protein 2, At the same time, β-sitosterol can inhibit the polymerization of tubulin in SiHa cells, and it gradually increases with the prolongation of the action time.Conclusion β-sitosterol can decrease the expression of tubulin α and microtubule-associated protein 2 in SiHa cells, and inhibit the polymerization of microtubules in SiHa cells, which suggests that β-sitosterol has a certain anti-microtubule effect, which may be an important reason for the growth inhibition of SiHa cells.

Objective to investigate the effect of arginine-glycine-aspartic acid (RGD) peptide-containing K16RGDSPC modified polylactide-co-glycolide (PLGA) - [ASP-PEG] scaffold material on the adhesion, proliferation and osteogenic differentiation of bone marrow stromal stem cells (BMSCs) on their surfaces.Methods K16GRGDSPC was synthesized by solid phase synthesis and identified by mass spectrometer and high pressure liquid chromatography.K16GRGDSPC was grafted to PLGA- [ASP-PEG] scaffold material with crosslinking agent Sulfo-LC-SPDP, and the grafting reaction was detected by XPS spectrum.The modified PLGA- [ASP-PEG] scaffold material was compounded with rabbit BMSCs in osteogenic induction medium, and the unmodified scaffold material was used as control. The changes of adhesion and proliferation of BMSCs were detected by precipitation method, microtubule sucking method, MTT method and Coomassie brilliant blue method, respectively. Alkaline phosphatase (ALP) kit was used to detect the expression of ALP in BMSCs, and the expression of ALP, osteocalcin (OCN), osteopontin (OPN) and type Ⅰ collagen was detected by RT-PCR. The expression of core binding factor a1 (Cbfa1) was detected by immunofluorescence staining, and the differentiation of BMSCs into osteoblasts was observed. Results XPS pattern confirmed that RGD peptide-containing K16GRGDSPC was successfully grafted to the surface of PLGA- [ASP-PEG]. The adhesion and proliferation of BMSCs on the surface of PLGA- [ASP-PEG] were significantly improved, and the expression of osteogenic markers (ALP, OCN, OPN, type Ⅰ collagen and Cfba1) was significantly higher than that of the control group (P<0.05). Conclusion RGD peptide-containing K16GRGDSPC can promote the adhesion and proliferation of BMSCs on the surface of bone matrix and induce their differentiation into osteogenesis.

Objective to investigate the regulatory effects of neutrophils (PMN) on dendritic cells (DC) during the immune response of T helper (Th) 1 cells. Methods LPS-stimulated neutrophils (LPS-PMN) and immature dendritic cells (imDC) were co-cultured, CD40 and CD86 on DC surface were detected by flow cytometry, and interleukin-12 (IL-12) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The DC co-cultured with LPS-PMN (PMN-DC) was purified and cultured with FITC-labeled ovalbumin (FITC-OVA) to detect its phagocytosis function. PMN-DC was co-cultured with DO11.10T cells and OVA17 peptide, the number of viable cells was counted, and interferon-γ (IFN-γ) and interleukin-4 (IL-4) were detected by intracellular staining. Results LPS-PMN could stimulate the upregulation of CD40 and CD86 in imDC and secrete IL-12 and tumor necrosis factor (TNF) -α.

Objective to investigate whether adult myoblasts can be transdifferentiated into neural precursors. Methods Myoblasts were isolated and cultured from 6 adult normal temporal muscle specimens. At the third passage, serum-free culture medium containing basic fibroblast growth factor, epidermal growth factor and leukemia inhibitory factor was added and induced for 2 weeks. The induced cells were identified by immunocytochemistry and analyzed by reverse transcription polymerase chain reaction (RT-PCR). The induced cells and human myoblasts were transplanted into the brains of 12 rats with cerebral ischemia, respectively, and the survival and differentiation of implant cells were identified by immunohistochemistry. Results the myoblasts began to aggregate into spheres after 7 days of induction and grow in suspension. The spheres were passaged every 14 days on average. All the spheres expressed nestin. Under the differentiation conditions, (36±6) % of the cells expressed microtubule-associated protein 2, (44±5) % of the cells expressed glial fibrillar acid protein, (17±4) % of the cells expressed galactocerebroside; The induced cells expressed microtubule-associated protein 2 and glial fibrillary acidic protein after transplantation into rat brain, while the control myoblasts did not express the above markers. Conclusion Human myoblasts can transdifferentiate into neural precursor cells under certain induction conditions in vitro.

Objective to compare the gene expression profiles of primary breast cancer and paired lymph node metastasis to screen the gene associated with breast cancer metastasis. Methods Firstly, the differentially expressed gene fragments of primary breast cancer and its paired lymph node metastasis were screened by mRNA differential display (mRNA DD) technique, and the differential gene fragments were cloned, sequenced and compared with GenBank; Then using isotope-labeled reverse dot blot hybridization and fluorescence-labeled gene chip hybridization methods to verify and screen the obtained differential genes; real-time quantitative reverse transcription polymerase chain reaction (real-time RT-PCR) was used to detect the mRNA expression of differentially expressed genes in seven breast cancer cell lines with different biological behaviors.Results the gene expression profiles of lymph node metastasis and primary cancer of breast cancer were highly consistent, but there were also a few differentially expressed genes; Sixteen differential genes were obtained by mRNA DD screening, including four unknown genes, which were registered in GenBank with the sequence numbers BG518428, BG518429, BM005520, BM005521, four unknown functional genes with known sequences and eight known genes.Among them, kinetin-like DNA binding protein (KNSL4) and dihydropyrimidine dehydrogenase (DYPD) were confirmed to be down-regulated in metastatic carcinoma compared with the primary carcinoma by isotope-labeled reverse dot blot hybridization and gene chip hybridization. The expression of KNSL4 mRNA in seven breast cancer cell lines with different biological behaviors showed a downregulated trend with the enhancement of cell metastasis capacity.Conclusion The similarity of gene expression profiles between primary breast cancer and paired lymph node metastatic oncology confirmed that lymph node metastatic carcinoma is a metastatic subclone of its primary carcinoma, and the differentially expressed genes of the two may be related to the metastatic phenotype of the cells; KNSL4 and DYPD are the genes with down-regulated expression in metastatic carcinoma confirmed by all three methods, and they are potential metastatic related genes of breast cancer; KNSL4 is associated with the biological behavior of breast cancer cell metastasis, suggesting that it may play an important role in breast cancer metastasis.

Objective to systematically evaluate the clinical effect of a new generation shoulder prosthesis-Univers 3-D prosthesis replacement in the treatment of glenohumeral osteoarthritis.Methods 74 cases of glenohumeral osteoarthritis were treated with UNIVERS 3-D shoulder prosthesis replacement, with an average follow-up of 31.9 months (12-74 months). The clinical follow-up included: functional assessment and X-ray photography. Results the Constant score was 68.40±16.91 after treatment of OA with UNIVERS3-D prosthesis replacement, which was 2 times higher than that before operation. Excluding age and sex differences, the functional correction score was 45.29%±19.87% before operation and 92.27%±22.53% after operation. The average subacromial distance was 10.91 mm ± 3.03 mm; The height and width of the humeral head were 5.71 mm ± 4.32 mm and 53.34 mm ± 4.82 mm, respectively. 36.96% had signs of humeral head necrosis; 8.69% had signs of cystic degeneration in the humeral head; 39.13% had osteophyte formation around the glenoid side of the scapula, and the spacing between glenohumeral joints increased significantly after operation; The subluxation of glenohumeral joint was effectively corrected before operation, and no interarticular impingement sign was found. The subacromial distance was significantly widened after operation compared with that before operation. The height and width of the humeral head were reconstructed after operation. The inclination angle of head-neck-trunk of the prosthesis changed continuously from 126 to 142°. No obvious change was found in the angle of head-neck-trunk of the humeral prosthesis after operation, and there was no separation and dislocation of the head-neck-trunk prosthesis. The incidence of X-ray lucid zone around the humeral shaft prosthesis was 5.41%, and the scores were all less than 1. The most frequent areas of lucid zone were D, c and e. No X-ray lucid zone was found around the glenoid prosthesis, and the complication rate was 12.2%. Conclusion UNIVERS 3-D shoulder prosthesis replacement in the treatment of glenohumeral osteoarthritis can reconstruct the anatomical relationship of glenohumeral joint, and the postoperative function is significantly improved compared with preoperative. The incidence of lucid zone and complication rate are low, and the clinical effect is excellent.

Objective to investigate the influencing factors of postoperative crisis in patients with generalized myasthenia gravis (MG). Methods A case-control study was performed on the influencing factors of postoperative crisis in 176 patients with generalized myasthenia gravis (MG). Postoperative crisis occurred in 36 patients. The research items included: sex, age at the time of operation, course of disease at the time of operation, pathological type of thymus, site of involvement at the time of operation, clinical classification at the time of operation (Osserman classification), history of preoperative infection (within 1 month before operation), history of preoperative crisis (within 1 month before operation), preoperative dosage of pyridostigmine (mg/d), preoperative hormone use, duration of operation (min), amount of bleeding during operation (ml), and pleural damage during operation. Results Univariate analysis showed: preoperative swallowing muscle involvement (OR =8.494, P =0.001), preoperative crisis history (OR =5.667, P =0.000), thymoma (OR =2.147, P =0.047), type III and IV (OR =4.549, P =0.000), preoperative infection history (OR =3.30, P =0.038), preoperative dosage of pyridostigmine Conclusion Preoperative swallowing muscle involvement, preoperative infection history, preoperative crisis history and preoperative pyridostigmine dosage are the independent influencing factors of postoperative crisis. Make full preparation before operation to reduce the occurrence of postoperative crisis.

A 22-year-old male was diagnosed as "central diabetes insipidus; suspected micro-intrasellar germ cell tumor" in Peking Union Medical College Hospital due to polydipsia and polyuria for 2 years. The symptoms improved significantly after treatment with diabetes insipidus and micagulation. Three months ago, the patient developed headache, fatigue and decreased visual acuity in both eyes. For further diagnosis and treatment, he was admitted to the Department of Neurosurgery of the First Affiliated Hospital of Harbin Medical University in March 2006. Physical examination: clear consciousness, hypopituitary face, scarce beard and pubic hair, binocular visual acuity 0.5, hemianopia in the temporal field of both eyes, normal fundus, cerebrospinal fluid and serum tumor markers (chorionic gonadotropin, alpha-fetoprotein, carcinoembryonic antigen and ferritin) were all normal.

The middle and lower slope area and its corresponding ventral brainstem area are deeply located and anatomically complex, and have always been one of the most challenging intracranial surgical areas, which is called "inaccessible area" [1-3]. The preauricular subtemporal-inferior temporal fossa approach is one of the main surgical approaches in this area [4, 5]. This approach exposes the middle and lower slope area and its corresponding ventral brainstem area from the anterolateral side of the skull base, and has the advantages of widely resecting the skeletal structure of the middle skull base, without facial nerve displacement and labyrinth exposure, avoiding the obstruction of many cranial nerves, and avoiding excessive brain traction. However, with the development of minimally invasive neurosurgery, this approach reveals the defects of large surgical trauma and more ineffective exposure. In addition, in order to reveal the internal carotid artery in the petrous segment, mandibular condyle resection has complications such as temporomandibular joint malocclusion, occlusion and chronic pain after operation. In this experiment, this approach is combined with the keyhole concept, and it is improved to reduce the approach trauma and ineffective exposure, while preserving the integrity of temporomandibular joint, It did not affect the exposure and displacement of internal carotid artery in petrous segment, and the exposure of middle and lower slope area and ventral structure of brain stem.

A 66-year-old male was admitted to the hospital on May 17, 2004 for adjusting blood glucose due to "polydipsia and polyuria for 18 years, numbness of both lower limbs and blurred vision for 6 years". In 1986, he was diagnosed with diabetes in a local hospital due to thirst, polydipsia and polyuria. He had taken oral hypoglycemic drugs such as Eupidron, Metformin and Mepida, and his fasting blood glucose was controlled at about 7 mmol/L and postprandial blood glucose was controlled at about 9 mmol/L. Six years ago, he developed blurred vision in both eyes and numbness of both lower limbs. He started insulin therapy: Novolin R was injected subcutaneously three times a day before meals and Novolin N was injected once a day before going to bed, and the insulin dose was adjusted according to blood glucose. At first, his blood glucose was controlled at about 7 mmol/L and postprandial blood glucose was controlled at about 9 mmol/L. In recent 4 years, his blood glucose was gradually increased, fasting blood glucose was controlled at about 9 mmol/L and postprandial blood glucose was controlled at about 14 mmol/L. He gradually increased his body weight, and denied his history of diabetic ketoacidosis. He had a history of hypertension for 4 years. He took Xintongding, compound antihypertensive tablets, Luo Blood pressure controlled at 150-160 mm Hg/100-110 mm Hg (1 mm Hg =0.133 kPa).

Autologous chondrocyte transplantation and cartilage tissue engineering provide a good way for the repair of articular cartilage injury. As an ideal source of seed cells for cell transplantation and cartilage tissue engineering, the biological characteristics of chondrocytes have been studied by many scholars at home and abroad, but many biological mechanisms are still unclear. At the same time, the research of cell membrane ion channels has become a hot spot with the development of patch clamp technology, molecular biology, morphology technology, immunology and other technologies. However, the research on ion channels of articular chondrocytes is limited, so it is necessary to deeply understand the research status of this field.

The growth and infiltration of solid tumor depends on the formation of neovascularization in tumor, and the establishment of intravascular system in tumor can be carried out in many ways. Previous studies have suggested that the endothelial cells that constitute tumor blood vessels may be derived from endothelial precursor cells of host bone marrow, hemangioblasts or already formed endothelial cells of blood vessel wall. Mosaic blood vessels and pseudo-angiogenesis enrich the theory of tumor neovascularization, but their theory that blood vessels can be formed in the absence of endothelium, which challenges the origin of traditional tumor vascular endothelial cells.

white matter lesions (WML), including paraventricular lesions and deep subcortical lesions, are common in senile brain tissues, increase with age and are closely related to cognitive impairment. WML showed low density in CT, and high density in T2-weighted phase and proton phase in MRI. Histological studies showed that the white matter of patients with Binswanger disease and Alzheimer disease (AD) showed myelin thinning, axon loss, oligodendrocyte loss, astrocyte proliferation and arteriosclerosis, etc. It was inferred that it may be related to cerebral ischemia, but the toxic effect of soluble β-amyloid protein could not be ruled out to cause nerve damage.

A large number of epidemiological and laboratory studies have confirmed that human papillovirus (HPV) infection is a necessary but not a sufficient condition for the pathogenesis of cervical cancer [1,2]. In recent years, some factors related to gene susceptibility of the body have been thought to be involved in the pathogenesis of cervical cancer. Fas gene is a gene related to apoptosis and belongs to the tumor necrosis factor receptor superfamily. The promoter region-670 of Fas gene is located in the nuclear transcription element GAS, and there is an A/G polymorphism at this site. The DNA sequence changes corresponding to this polymorphism can affect the expression of Fas gene. The abnormal apoptosis mediated by Fas/FasL pathway is related to the occurrence of many diseases, which can make tumor cells escape immune supervision and lead to the occurrence of tumor [3]. There are reports abroad about the relationship between Fas-670 polymorphism and cervical cancer and other tumors [4-6], and some scholars in Taiwan Province of China have also reported the relationship between Fas gene polymorphism and cervical cancer [7], but the reports are different, and the relationship between Fas gene polymorphism and cervical cancer has not been found in mainland China Therefore, we studied the relationship between single nucleotide polymorphism of Fas-670 site and susceptibility to cervical cancer in Han women, which is reported as follows.

Intraplacental choriocarcinoma is a very rare case in clinic. Generally, the possibility of primary choriocarcinoma of the placenta is suspected only after the clinical symptoms occur in the parturient or the clinical symptoms are found in the child. At present, there are no case reports in China, and only individual cases are reported in international literature, and often there are no complete clinical and pathological data. In 2005, Peking Union Medical College Hospital treated a case of choriocarcinoma in both mothers and infants, and the mother and infant were cured at the same time. The report is as follows.

thyroid receptor associated protein 220 (TRAP220) forms a complex with steroidal hormone nuclear receptor in a ligand-dependent form, which is an important link in the physiological role of thyroid hormone and estrogen in the brain [1-3]. We investigated the expression of TRAP220 in the brain tissues of patients with NFDA1A with drug-resistant temporal lobe epilepsy from the gene and protein levels, and explored its clinical value.

tissue-type plasminogen activator (tPA) is an ideal thrombolytic drug, and its gene has been successfully transduced into vascular endothelial cells, smooth muscle cells and vascular tissues. However, the functional expression of tPA gene transduced into neonatal rat myocardial fibroblasts has not been reported. In this study, a novel pseudotype retrovirus vector was used to mediate the transduction of tPA gene into neonatal rat myocardial fibroblasts, in order to observe the functional expression of myocardial fibroblasts after transgenic transduction, and provide experimental basis for further research on antithrombotic gene therapy after cardiovascular surgery.

The concept of apical pleural tumor is more extensive than that of superior pulmonary sulcus tumor, but the relationship between its adjacent structures is similar to that of superior pulmonary sulcus tumor. The anatomical structure of the pleural apex is complex, so once tumor occurs, it is easy to invade the adjacent structure, which brings great difficulty to surgical treatment. From January 2000 to February 2006, 10 cases of pleural apex tumor were surgically treated in our hospital. The characteristics and results of surgical treatment are reported as follows.