Volume 86  ·   Issue 21  ·  2006

Methods Type 2 adeno-associated virus recombinants were replicated and packaged by AAVMaxTM packaging system, purified and titer tested, and then transfected with different transfection indexes (MOI) in New Zealand rabbit articular chondrocytes in vitro. The transfection effect and the relationship between transfection effect and time were observed. The transfection effect was confirmed by inverted fluorescence microscope, and the transfection effect was detected by flow cytometry, including the percentage of cells expressing green fluorescence signal and the average fluorescence intensity. The maximum percentage of green fluorescence signal expressed spontaneously by untransfected chondrocytes was 4.76% and the maximum fluorescence signal intensity was 1.04 as blank control. The dose-response curve between MOI (v.g. /cell) and transfection rate showed a linear positive correlation when MOI was less than 105 v.g. /cell, and the transfection rate did not increase significantly when MOI was more than 105 v.g. /cell. The expression of green fluorescence signal reached its peak on the 7th day of transfection, and the percentage of chondrocytes expressing green fluorescence signal and fluorescence intensity showed a decreasing trend with the prolongation of transfection time and the increase of passage times, but the expression of green fluorescence protein could still be detected on the 56th day after transfection. Enhanced green fluorescent protein is a good reporter gene to observe the effect of adeno-associated virus type 2 recombinant transfection into articular chondrocytes.

Objective to screen serum protein molecular markers of hypopharyngeal carcinoma and establish a decision-making diagnostic model. Methods The sera of 48 patients with hypopharyngeal carcinoma, of which 36 were modeling group, 12 were blind validation group and 52 normal controls, of which 36 were modeling group and 16 were blind validation group, were determined by surface-enhanced laser dissociation ionization time-of-flight mass spectrometry using weak cation exchange chip (WCX2) and fixed metal affinity chip (IMAC3). The difference of protein profiles between hypopharyngeal carcinoma and normal human serum was analyzed, and a classification tree model was established and verified by dyspharyngeal method. Results A total of 11 differential protein peaks were detected by WCX2 chip and 19 differential protein peaks were detected by IMAC3 chip in the range of mass-to-charge ratio (M/Z) 2~50. The decision tree model was automatically selected by 7796, 4216, 5927 and 5361 (M/Z). The sensitivity was 94.44% (34/36) and the specificity was 88.89% (32/36). The sensitivity of blind validation was 91.67%, The specificity is 81.25%. Conclusion the marker proteins related to hypopharyngeal carcinoma can be screened from patients' serum by using this method. The established decision tree model may have important clinical significance for early detection and early diagnosis of hypopharyngeal carcinoma.

Objective to study the expression characteristics of transforming growth factor-β 1 (TGF-β 1) and basic fibroblast growth factor (bFGF) in the cartilage of the apical convex and concave inferior articular processes of the most severe juvenile idiopathic scoliosis (AIS). Methods Twenty-two patients with AIS were studied, and 18 patients with congenital scoliosis (CS) were used as controls. The pathological changes of the articular processes and the distribution characteristics of TGF-β 1 and bFGF in the articular processes were observed by HE staining, immunohistochemistry and in situ hybridization methods. The obtained immunohistochemistry and in situ hybridization images were input into the image analysis system for semi-quantitative analysis. Results the expression characteristics of TGF-β 1 and bFGF in AIS and CS were basically similar. The expression characteristics of TGF-β 1 and bFGF in the concave side of the apical vertebra were higher than that in the convex side (165.4±59.2 vs 128.1±50.6, 168.3±46.8 vs 126.5±47.0; 96.4±28.4 vs 76.6 The expression of TGF-β 1 and bFGF in the concave side of the apical vertebrae in AIS may be compensated by the reconstruction of the intercellular matrix of the articular processes caused by the abnormal biomechanics after spinal deformity, and abnormal compressive stress may play a very important role.

Methods 3T3-L1 preadipocyte line and C2C12 myoblast line were induced to differentiate into 3T3-L1 adipocytes and C2C12 skeletal muscle cells, respectively. The two cells were treated with 10-9mol/L testosterone for 4, 8, 12, 24 and 48 h, respectively, and treated with different concentrations of testosterone (10-12~10-5mol/L) for 24 h. The expression changes of insulin receptor substrate 1 (IRS-1) and glucose transport carrier 4 (GLUT4) in the two cells were detected by Western blotting. Results The expression of IRS-1 and GLUT4 in the two cells increased gradually with the prolongation of the treatment time of 10-9mol/L testosterone, and the expression of IRS-1 decreased after reaching the peak at 12 h. The peak values of 3T3-L1 adipocytes and C2C12 skeletal muscle cells at 12 h were (1.42±0.42) times and (1.53±0.14) times of the 0 h control, respectively, both P<0.05. The expression of GLUT4 in 3T3-L1 adipocytes and C2C12 skeletal muscle cells increased (4.23±0.27) fold and (3.16±0.15) fold after 10-9mol/L testosterone treatment, respectively, P<0.05. The expression of GLUT4 in C2C12 skeletal muscle cells increased with the increase of testosterone concentration [the expression of GLUT4 in C2C12 skeletal muscle cells after 10-7mol/L testosterone treatment was (2.99±0.15) fold, P<0.05, but the increase trend was not obvious after 10-7mol/L. In 3T3-L1 adipocytes, the expression of GLUT4 peaked at 10-11mol/L [increased (2.58±0.02) fold, P<0.05], and then decreased with the increase of testosterone concentration. Conclusion Testosterone affects the expression of insulin signal transduction molecules in both dose and time.

Objective to investigate the characteristics of peritubal capillary (PTC) damage in chronic aristolochic acid nephropathy (CAAN) and the effect of hypoxia induced by PTC damage on the progression of CAAN.Methods 54 Wistar rats were randomly divided into two groups: 30 rats in the Guanmutong group were given Guanmutong decoction by gavage for 8 weeks, and 10 rats were sacrificed at the 8th, 12th and 16th weeks respectively; In the control group (24 rats), 8 rats were sacrificed at the 8th, 12th and 16th weeks. Blood, urine and kidney tissue samples were collected for biochemical, pathological, immunohistochemical and immunoblotting examinations. Results From the 8th week, the renal function of the rats in the Guanmutong group gradually deteriorated, the urine protein increased, the damage of PTC aggravated, the density of PTC decreased, the expression of vascular endothelial growth factor (VEGF) gradually decreased, but the expression of hypoxia-inducible factor 1 α (HIF-1 α) gradually increased. (2) The PTC density in the control group was (42.80±4.49) /0.13 mm2, and that in the Guanmutong group was (8.10±2.28) /0.13 mm2 (P<0.01); (3) The value of VEGF-IOD was (26.94±9.34) ×103 in the control group and (2.78±0.78) ×103 in the Guanmutong group (P<0.01). The renal interstitial area decreased at the 8th week compared with the control group, gradually increased at the 12th week, and significantly increased at the 16th week.Conclusion There are severe PTC damage and hypoxia in CAAN rats, and ischemia and hypoxia are related to the progression of CAAN.

Objective to observe the effect of purified Yunnan cobra venom factor (Y-CVF) on acute humoral rejection after presensitized rat heart allograft. Methods BN rats to Lewis rats were subjected to ectopic heart transplantation after three consecutive skin transplantation. Fifteen pairs of rats were divided into two groups by random number method. The experimental group (n =8) was given Y-CVF 80 μ g/kg intravenously 24 h before heart transplantation; In the control group (n =7), the survival time of transplanted heart was observed, the type of rejection was examined by pathology after heart arrest, and the deposition of IgG and complement C3 was observed by immunohistochemical staining. Results the antibody titers of anti-BN rats increased from 0 to 1:1028~1:2056 after presensitization in Lewis rats. The survival time of transplanted heart in the control group was 12.71 h ± 13.94 h, and the survival time of transplanted heart in the experimental group was 99.50 h ± 38.72 h, and the difference was statistically significant compared with that in the control group (t =5.599, P<0.01). The pathological examination results confirmed that there was no acute humoral rejection in the experimental group, only acute cell rejection characterized by massive mononuclear lymphocyte infiltration was observed, and acute humoral rejection characterized by thrombosis in small blood vessels was observed in the control group. Immunohistochemical IgG staining was positive in the experimental group and the control group, and C3 staining was positive in the control group, while the experimental group was negative. Conclusion Y-CVF can overcome the acute humoral rejection of presensitized rat heart allotransplantation

Objective to investigate the value of physiological and surgical invasiveness score (POSSUM) in predicting the incidence of postoperative complications in elderly patients with esophageal and cardiac carcinoma in thoracic surgery. Methods POSSUM score was used to predict postoperative complications in 167 elderly patients with esophageal and cardiac carcinoma in thoracic surgery, and compared with the actual values. Results 78 patients had postoperative complications, and the physiological score of the complication group was significantly higher than that of the uncomplication group (16.75±0.4 vs 15.4±0.3, P<0.001). The number of postoperative complications predicted by POSSUM score was 70 cases, and the actual number of complications was 78 cases, with no statistical significance (P =0.378); There were 94 complications in 78 patients, including 12 cases of hypertension, 37 cases of arrhythmia, 6 cases of phlebitis, 5 cases of asthma, 3 cases of fever (excluding postoperative absorbed heat), 3 cases of incision infection and 3 cases of incision liquefaction, 2 cases of pneumonia, pneumothorax, respiratory failure, anastomotic fistula, acute gastric dilatation, postoperative hemorrhage, empyema and deep vein thrombosis, 1 case of heart failure, pulmonary infarction, atelectasis, septic shock, chylothorax, cerebral infarction, infusion reaction, urinary tract infection and psychiatric symptoms.

Objective to investigate the configuration and variation of colonic vessels and their influence on the blood supply of colonic segment replacing esophagus. Methods the origin, direction, branch, configuration, distribution range, pulsation strength of colonic vessels, the integrity of the marginal vessels in hepatic flexure and splenic flexure area were observed in 582 cases of colonic esophageal replacement surgery, and the best transplantation of colonic segment blood supply vessels was screened. Results 97.3% of left colonic artery originated from inferior mesenteric artery with single branch or common trunk, and only 0.7% were absent; The middle colonic artery originated from the superior mesenteric artery alone in 77.8%, the accessory middle colonic artery existed in 6.2%, the interference with the right colonic artery in 7.8%, and the absence in 8.2%; The right colonic artery originated from the superior mesenteric artery alone in 39.7%, interfered with the middle colonic artery in 23%, interfered with the ileocolic artery in 28%, and was absent in 9.8%. The marginal vascular arch in the splenic flexure area was intact in 96.8%, and the marginal vascular arch in the hepatic flexure area was intact in 88.7%. The lateral and accessory marginal vascular arch (Riolan arch) only existed in 7.6% of the cases. Conclusion The colonic vascular configuration in Chinese is similar to that in foreign countries, but the absence rate, number and distribution of vascular branches in the colonic margin are different. The left colonic artery is the best donor vessel for transplantation, followed by the middle colonic artery.

Objective to evaluate the application value of circular gastrointestinal stapler in surgery of esophageal and cardiac carcinoma. Methods 3322 cases of esophageal and cardiac carcinoma were mechanically anastomosed by circular stapler from August 1980 to September 2004, including 2642 males and 680 females; There were 2312 cases of esophageal cancer and 1010 cases of cardia cancer; There were 102 cases of esophagogastric and cervical anastomosis, 147 cases of thoracoparietal anastomosis, 1838 cases of supraarchal anastomosis and 1235 cases of inferior archal anastomosis.Results 18 cases (0.54%) had postoperative anastomotic leakage, including 11 cases of intrathoracic anastomotic leakage and 7 cases of cervical anastomotic leakage.9 cases (81.8%) died of intrathoracic anastomotic leakage.66 cases (2.0%) had postoperative anastomotic stenosis and 36 cases (1.1%) had instrumental failure during anastomosis.Conclusion Circular digestive tract stapler is safe and reliable in upper digestive tract reconstruction, and mechanical anastomosis is an effective technique to reduce postoperative anastomotic leakage.

The patient was a 30-year-old woman with 3 pregnancies and 0 deliveries. She was admitted to the hospital on July 25, 2005 due to "5 months of menopause, chest tightness and holding breath for 2 months, and anuria for 1 month". Her menstruation was regular, 4~6/30 days, moderate amount, dysmenorrhea (-), and her last menstruation was 5 months ago. After 3 months of menopause, she felt chest tightness and holding breath, which worsened in the past month. She could not lie down at night, and her daily urine output was less than 100ml.

Neurosurgery has experienced the process of enlarging the surgical incision to make the light illuminate the deep intracranial part, ensuring that the surgeon and assistant have sufficient illumination for naked eye surgery → glasses-type surgical magnifying glass → surgical microscope [1]. The design and perfection of the skull base approach in microneurosurgery has exposed and resected the lesions involving the midline of the brain and the skull base that were previously considered inaccessible. On this basis, the approach with the keyhole concept has been used in those small, asymptomatic or early-detected lesions and vascular diseases located in ideal sites in order to reduce unnecessary surgical trauma. Although the application of neurosurgical keyhole technology has been applied abroad for nearly 20 years, and the introduction into China has a history of 5 years [2, 3]. However, due to the limitation of bone hole and the risk of surgery, and the misunderstanding of keyhole concept, there has been a debate about keyhole surgery. On the one hand, it actively advocates to alleviate the pain of patients, constantly improve the surgical technology, and move towards the new height of microneurosurgery keyhole technology [4]; On the one hand, it emphasizes the safety of patients and the operative effect, avoids the adverse consequences and serious complications brought by the development of new technology, and considers that it is a great mistake to apply the theory of keyhole magnification to neurosurgery [5]. There is no applied technology in neurosurgery that has such great differences in the introduction and development. In order to understand and apply this technology well, the following aspects are reviewed.

Peroxisome-activated receptors (PPAR) constitute the nuclear hormone receptor family [1], belonging to the large family of steroid receptors, and participate in various physiological activities, such as steroid synthesis, blood vessel growth, tissue remodeling, cell cycle, apoptosis, lipid metabolism, etc. The activation or inhibition of PPAR can regulate the above physiological activities, and many factors can activate or inhibit PPAR. Therefore, it is very important to study the biological function of PPAR, and the study of PPAR ligands (activators) has a broad clinical application prospect. The role of PPAR in reproductive biology has also attracted great attention, but the biological function of PPAR in ovary is rarely studied. Therefore, the biological characteristics of PPAR and its role in ovarian cells are summarized as follows.

The mechanical properties of chondrocytes can significantly affect the physiological function of articular cartilage. The study of the mechanical properties of chondrocytes not only helps to clarify the role of biomechanical factors in the pathogenesis of osteoarthritis, but also provides new ideas and technical methods for the study of related mechanical factors in cartilage tissue engineering, and provides relevant theoretical basis for the treatment of cartilage injury by tissue engineering.

How to prolong life span has always been a hot topic in the scientific community. Although studies have shown that long-term calorie intake can prolong the life span of rodents as early as 1986, it is still unclear whether this way will affect the biological markers of human longevity and oxidative stress, and whether the body's metabolic rate can be reduced to a level beyond the expected weight loss. Recently, a team led by Dr. Eric Ravussin of the Plimpton Biomedical Research Center of Louisiana State University in the United States conducted a randomized controlled study. The results of the study were published in<JAMA, 2006, 295: 1539-1548.

Glycogen synthase kinase (GSK) -3 β is a serine/threonine kinase, which can phosphorylate many substrates, thereby exerting many important cellular functions such as regulating cell proliferation and apoptosis [1,2]. Previous studies have shown that GSK-3 β can promote the proliferation of ovarian cancer cells, but its mechanism is still unclear.

Intraspinal angiolipoma is a rare benign lesion, and 111 cases have been reported at home and abroad [1-5]. A total of 7 cases were treated in Beijing Tiantan Hospital from October 1992 to February 2005. The clinical characteristics, diagnosis and treatment of the angiolipoma are reported as follows.

Stage Ⅱ autologous bone grafting is the main method to eliminate bone defects after osteomyelitis debridement. Its shortcomings are staged surgery, limited donor bone volume, trauma and complications of donor site surgery, but allogeneic bone grafting has not been recognized yet. On the basis of the good curative effect of stage Ⅰ autologous cancellous bone grafting [1], we used stage Ⅰ autologous bone grafting or allogeneic bone grafting to treat chronic osteomyelitis respectively. The effects of the two treatment methods are compared as follows.