Volume 87  ·   Issue 17  ·  2007

Regenerative medicine is one of the most rapidly developing fields in biology and medicine in recent years. With the deepening of people's understanding, the increase of investment in science and technology and the application of a large number of high and new technologies, remarkable achievements have been made in basic research and clinical treatment of regenerative medicine, especially adult stem cells and regenerative medicine, which provides important theoretical and experimental basis for large-scale therapeutic application in the future.

Objective to investigate the predictive factors and predictive molecules of platinum combined with paclitaxel as first-line chemotherapy for advanced ovarian cancer. Methods the clinicopathological data of 42 cases of advanced ovarian epithelial cancer treated with platinum combined with paclitaxel as first-line chemotherapy were reviewed. The expression of p53 and PgP in ovarian cancer tissues was detected by immunohistochemistry, and the clinical complete remission rate and early relapse rate of first-line chemotherapy were calculated. The feasibility of age, residual lesion size, pathological type, differentiation degree, p53 and PgP expression as predictive factors or predictive factors of chemotherapy was analyzed. Results 24 cases (57%) had complete clinical remission, and 7 cases (17%) had early relapse. Univariate analysis showed that the complete remission rate of p53 positive patients (74%) was higher than that of p53 negative patients (43%); The early recurrence rate of patients with residual lesions<2cm (4.2%) was lower than that of patients with residual lesions ≥2cm (33%). Multivariate analysis suggested that residual lesions ≥2cm and Pgp positive were independent prognostic factors for early recurrence. Conclusion The size of residual lesions, p53 expression and Pgp expression can be used as predictive factors for the efficacy of platinum combined with paclitaxel chemotherapy.

Objective to investigate the clinical characteristics and recurrence pattern of recurrent ovarian mature teratoma. Methods The clinical data of 20 cases of ovarian mature teratoma (recurrence group) who recurred after conservative surgery were retrospectively analyzed and compared with 40 cases without recurrence (control group) who operated at the same time. Results The recurrence rate of ovarian mature teratoma after conservative surgery was 2.5%. The average recurrence time was (8±7) years. Young patients and bilateral or multilocular cysts were more likely to recurrence. There was no statistically significant difference between the recurrence group and the control group in the number of pregnancies and births, cyst size, surgical method and whether it ruptured during operation. Conclusion The follow-up should be strengthened for young patients and bilateral or multilocular teratoma. Laparoscopic surgery should be used as much as possible for teratoma surgery. There is no need to routinely perform contralateral ovarian anatomy during unilateral teratoma surgery.

Objective to investigate the clinical diagnosis and treatment of perineal endometriosis. Methods 30 cases of perineal endometriosis (endometriosis) were retrospectively analyzed and followed up for 0.5~13 years. Results Perineal endometriosis accounted for 0.32% of endometriosis in the same period in hospital, 0.0065% of perineal endometriosis occurred in vaginal delivery, and the incubation period of onset was from 4 months to 13 years. Perineal endometriosis all showed periodic and painful nodules in perineal wound, with no significant increase in CA125 value, and the patients with complete resection could be cured. 5 cases (16.7%) were complicated with pelvic endometriosis. Conclusion Correct diagnosis can be made according to typical medical history and physical examination. Perineal endometriosis can be complicated with pelvic endometriosis, and surgical resection is the main treatment. The patients without anal sphincter can be completely resected by surgery, and the recurrence rate is lower than those with anal sphincter involvement. Preoperative evaluation of whether anal sphincter involvement is crucial.

Objective to investigate whether long-term continuous positive airway pressure (CPAP) ventilation can reduce blood pressure in patients with obstructive sleep apnea hypopnea syndrome (OSAHS). Methods the relevant literatures from 2000 to 2006 were retrieved from MEDLINE and the computer retrieval system of Chinese Journal Full Text Database. The observation subjects were limited to patients with OSAHS who were treated with CPAP for ≥4 weeks. The data of all clinical randomized controlled studies comparing the effects of CPAP and non-CPAP treatment on 24h blood pressure were meta-analyzed. Results A total of 471 patients from 7 studies were included in the meta-analysis, of which 3 studies supported that CPAP could reduce 24h mean diastolic blood pressure in patients with OSAHS. The results of the meta-analysis showed that long-term CPAP treatment can reduce the 24-h mean diastolic blood pressure in patients with OSAHS weighted mean difference: -1.78 (95% CI: -3.34, -0.22)]. Conclusion Long-term CPAP treatment can reduce the 24-h mean diastolic blood pressure in patients with OSAHS.

Objective to evaluate the imaging and clinical effects of multi-segment cervical Bryan Disc artificial Disc replacement. Methods The clinical data of 20 cases of multi-segment Bryan Disc artificial Disc replacement were retrospectively analyzed, including 18 cases with double segment and 2 cases with three segments. The measurement of replacement segment mobility, motor segment height, adjacent segment mobility, JOA score, JOA improvement rate, NDI score and Odom's standard were evaluated. Results The average mobility of replacement segment was 86.2% before operation, and the average mobility of cervical spine was 87.3% before operation. The average JOA score was (12.7±3.0) before operation and (16.1±1.6) after follow-up, with statistically significant differences (P<0.05), and the average JOA improvement rate was 80.1%. The follow-up NDI score was (8.7±7.5). According to Odom's standard, 10 cases (50%) were very good; 8 cases (40%) were better; Conclusion the early follow-up results and imaging results of multi-level cervical Bryan Disc artificial Disc replacement are satisfactory, and the degeneration of adjacent vertebral body can be reduced while ensuring cervical vertebral mobility and cervical lordosis. On the premise of strictly mastering the surgical indications, cervical Disc replacement is a good alternative to anterior cervical bone grafting fusion.

Objective to explore the application of model surgery under the guidance of three-dimensional cranial model technique in the repair of mandibular defects by vascularized autologous fibula graft.Methods Twenty-four patients with mandibular defects from 2001 to 2005 were treated with CT scan combined with rapid prototyping technique.Model surgery was performed on the three-dimensional cranial model.Personalized mandibular medical reconstruction titanium plate was prefabricated.The vascularized autologous fibula graft was used to repair mandibular defects and the curative effect.Results After the repair of mandibular defects caused by various causes guided by three-dimensional cranial model, the wounds healed in stage I. Six months after the operation, the facial shape was symmetrical, the opening was not restricted, the bite-jaw relationship on the healthy side was not deviated, the bilateral temporomandibular joint area was not tender,the oral mucosa and submandibular fistula were not found, and the lower limb movement was not obstructed.The oral panoramic film showed that the transplanted fibula and mandibular bone were completely bony healed.The three-dimensional CT scan showed that the mandibular shape was well restored.Conclusion The three-dimensional cranial model can truly reflect the lesion focus, It provides guidance for accurate diagnosis and determination of treatment plan, provides simulation platform for simulated surgery, and has a good auxiliary role in mandibular defect repair.

Objective to investigate the surgical indications and perioperative management of lung transplantation and combined cardiopulmonary transplantation in the treatment of Eisenmenger syndrome.Methods Five patients with congenital cardiac malformations associated with Eisenmenger syndrome (ES) were treated from September 2002 to March 2006. Among them, 2 patients with congenital cardiac malformations were atrial septal defect and 2 patients with ventricular septal defect. Another patient suffered from congenital heart disease with atrial septal defect and developed total heart failure after interventional atrial defect occlusion, and urgently underwent cardiopulmonary transplantation. Results Among the 4 patients with cardiac malformation repair and single lung transplantation at the same time, 1 patient died of primary graft failure 20 days after operation; One patient died of primary graft failure 23 days after cardiopulmonary transplantation. Three patients recovered and were discharged from hospital, and now they have survived for 5, 8 and 22 months respectively, with good quality of life. The pulmonary artery pressure decreased significantly and the results of blood gas analysis improved significantly after operation. Conclusion Single lung transplantation at the same time of cardiac malformation repair can effectively treat congenital cardiac malformation accompanied by Eisenmanger syndrome.

Objective to study the biological response of bone tissue to stress-strain parameters.Methods A micro-resistance strain gauge was pasted on the surface of the left ulna of rats, and the strain parameters of the left ulna were detected in vivo during normal walking.The mechanical biological model of rats was established by simulating the strain parameters in vivo with INSTRON material testing machine. Sixty rats were randomly divided into four groups, normal group, 5, 10 and 15 Hz groups, and each experimental group was divided into 2000 μ ε group and 3000 μ ε group; The right upper limb was loaded with the amplitude of 0.1 mm and half sine wave for 10 min/d for 2 weeks. The ulnar bone mineral density (BMD) of rats in each group was measured by dual energy X-ray bone mineral densitometry (DXA) before and after the experiment. Three-point bending test to test the mechanical properties of aggregate materials; Evaluation of bone matrix protein secretion by immunohistochemical staining; Morphological changes of bone tissue were observed by conventional HE staining, tetracycline hydrochloride and calcein double fluorescence labeling. Results in the 10 and 15 Hz groups, the secretion of osteocalcin and collagen type I was promoted in 2000 μ ε and 3000 μ ε groups, and two obvious yellow and green fluorescent bands were observed by double fluorescence labeling. The bone mineral density of the experimental group was significantly higher than that of the normal group, (102 ± 8) mg/cm2 ratio (113 ± 6) mg/cm2, P<0.05, but the mechanical properties of bone were significantly lower than that of the control group (P<0.05) and the normal group (P<0.01); There was no significant increase in bone mineral density in the 5 Hz group. Conclusion A biological model of bone mechanics in vivo rats was successfully constructed, and it was proved that short-term low amplitude and high frequency stress can promote bone growth within a certain strain range.

Objective to investigate the expression of K-Cl co-transporter 1 (KCC1) in cervical cancer tissues and its role in the occurrence and development of cervical carcinoma. Methods The expression and localization of KCC1mRNA and protein in 40 cases of cervical cancer, 10 cases of cervical intraepithelial neoplasia (CIN) and 10 cases of chronic cervicitis were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Western blot and immunofluorescence staining. Results The expression of KCC1mRNA and protein in cervical cancer tissues (gray ratio 1.29±0.17, 1.18±0.27, respectively) was significantly higher than that in chronic cervicitis (0.59±0.27, 0.33±0.18) and CIN tissues (0.50±0.28, 0.27±0.15), and the difference was statistically significant (all P<0.05). The expression of KCC1mRNA and protein in low and moderately differentiated cervical carcinoma tissues (gray ratio 1.39±0.12, 1.37±0.15, respectively) was significantly higher than that in highly differentiated cervical carcinoma tissues (1.16±0.14, 0.96±0.19), and the difference was statistically significant (P<0.05); The expression of KCC1 mRNA and protein in advanced cervical cancer tissues was not significantly different from that in the early group, and the distribution of KCC1 protein in cervical cancer and control tissues was localized on the cell membrane. Conclusion KCC1 gene may play an important role in the development of cervical cancer.

Objective to observe the inhibition of RASSF1A gene on the growth of esophageal carcinoma cell line EC9706. Methods the plasmid containing RASSF1A gene was transfected into EC9706 cells with deleted expression of RASSF1A to establish a stable transfected cell line. The cell activity and proliferation capacity were detected by MTT assay, the effect on cell cycle was detected by flow cytometry, and the tumor formation characteristics of transfected cells in vivo were detected by nude mice transplanted tumor assay. Results the expression of RASSF1A protein in EC9706 cells with stable expression of RASSF1A was increased compared with the control cells. The cell growth rate was slowed down (P<0.01); In the cell cycle, the proportion of G1 phase increased, and the proportion of S phase decreased (G1 phase: RASSF1A group: 68.53% ± 3.34%, empty plasmid group: 54.25% ± 4.61%, untransfected group: 53.41% ± 5.60%, S phase: RASSF1A group: (23.19±5.04) %, empty plasmid group: (31.81±2.05) %, untransfected group: 32.09%±1.99%) (P<0.01); Conclusion Exogenous expression of RASSF1A gene can significantly inhibit the growth of esophageal cancer cells in vitro and in vivo, suggesting that RASSF1A gene plays an important role in the development of esophageal cancer.

Methods 40 healthy Sprague-Dawley rats were randomly divided into four groups: sham control (sham) group, pulmonary ischemia-reperfusion (I/R) group (left hilum was blocked for 30 min and reperfused for 2 h), heme chloride (Hemin) group (given the inducer of heme oxygenase-1) and zinc protoporphyrin (ZnPP) group (given the inhibitor of heme oxygenase-1). The activity of superoxide dismutase (SOD) and plasma tumor necrosis factor (TNF-α) in lung tissues of each group were measured, and the wet/dry weight ratio (W/D) of lung tissues and the ultrastructural changes of lung tissues under electron microscope were observed. Results the wet/dry weight ratio (5.92±0.66) in Hemin group was lower than that in I/R group (7.55±0.66) (P<0.01), the activity of SOD (9.2±0.5) was higher than that in I/R group (2.8±0.4) (P<0.01), and the content of TNF-α (60.37±8.25) was lower than that in I/R group (452.26±22.59) (P<0.01 Conclusion the induction of heme oxygenase-1 can effectively protect the pulmonary ischemia-reperfusion injury.

Objective to investigate the effect of adenovirus vector transfection on rat grafted veins. Methods Wistar rats were used to interpose jugular vein graft in ipsilateral common carotid artery. The grafted veins were stained with HE and VG staining to observe the thickness of the intima of the blood vessels. The expression and proliferation of proliferating cell nuclear antigen (PCNA) in intima smooth muscle cells (SMC) were observed by immunohistochemical staining. The transfection rate of adenovirus was measured by inverted fluorescence microscope and excited by ultraviolet light. The levels of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Results the expression of GFP was higher 3 days after operation. The expression of GFP decreased at 10 days after operation; At 30 days after transplantation, there was almost no expression of GFP. Obvious intimal hyperplasia was observed in the experimental group and the control group at 10 days after transplantation. The intimal hyperplasia was reduced in the experimental group and the control group at 30 days after transplantation. Compared with the normal non-transplanted vein, the intimal hyperplasia was significantly decreased in the experimental group and the control group (P<0.05), and the intimal hyperplasia was slightly more obvious in the experimental group than the control group, but the difference was not statistically significant. There were very few PCNA positive cells in the normal vein smooth muscle cells in the experimental group, but the proliferation of PCNA positive cells in the intimal and media SMC increased and reached the peak at 10 days after transplantation. The PCNA positive cells in the medium SMC decreased significantly at 30 days after transplantation. Compared with the normal non-transplanted vein, the PCNA positive cells in the experimental group and the control group increased significantly (P<0.05). The PCNA positive cells in the experimental group were slightly more than the control group, but the difference was not statistically significant. The mRNA and protein expression of ICAM-1 and VCAM-1 in the normal vein were very little, but the positive expression of some cells in the control group appeared at 3 days after transplantation, and the most expression was observed at 10 days after transplantation The difference was statistically significant (P<0.05). The expression in the experimental group was higher than that in the control group, but the difference was not statistically significant compared with that in the control group. Conclusion Adenovirus vector can efficiently carry out gene transfer in vivo, and the action time is short. Adenovirus vector can promote the activation and increase of SMC in the vascular graft vein, intimal hyperplasia, ICAM-1, VCAM-1 expression, and inflammation and intimal hyperplasia after adenovirus transfection into the blood wall of the graft vein, which confuses the effect of the target gene.

Objective to investigate the inhibitory effect of tirazamine (TPZ) combined with phosphatidylinositol-3 kinase inhibitor (LY294002) on human ovarian cancer cell lines in vitro. Methods Under hypoxia and air conditions, TPZ alone and combined with LY294002 were applied to human ovarian cancer cell lines HO8910PM and OVCAR-3 at different drug concentrations, respectively. The inhibitory effect of TPZ on HO8910PM and OVCAR-3 was evaluated by tetramethylazolazole blue (MTT) colorimetric assay. The IC50s of TPZ combined with LY294002 were 22.7 μ mol/L and 31.5 μ mol/L for HO8910PM and 49.1 μ mol/L and 51.0 μ mol/L for OVCAR-3 respectively under hypoxia and air conditions. Conclusion TPZ has an inhibitory effect on human ovarian cancer cell lines, and the inhibitory effect is higher under hypoxia than under air conditions (P<0.01); LY294002 had a synergistic effect on TPZ. Compared with TPZ alone, the combination of LY294002 significantly reduced the IC50 of TPZ on cancer cells (P<0.01).

Objective to study the effect of pirh2 on the growth of lung cancer. Methods the expression of pirh2 in lung cancer was detected by immunohistochemistry; Short hairpin RNA (shRNA) interfering plasmids targeting pirh2 gene (psiRNA-Pirh2A and psiRNA-Pirh2B) were constructed, and a negative control group (psiRNA-Con) and an empty plasmid group (psiRNA-hH1) were designed to transfect A549 cells; The mRNA and protein expression of pirh2 were detected by real-time quantitative PCR and Western blot, respectively. The proliferation of A549 cells was detected by CCK-8 method; According to the results of CCK-8, a strongly inhibitory pirh2 interfering plasmid was selected for tumor formation experiment in nude mice. Results 42 cases (79.2%) of 53 lung cancer tissues were positive for pirh2; The cell growth of psiRNA-pirh2A and psiRNA-pirh2B groups was slower than that of the normal group (q values were 6.82 and 5.63, respectively, all P<0.05), while the cell proliferation ability of psiRNA-Con and psiRNA-hH1 groups was not statistically significant (t =6.27, P<0.05). The tumor tissue weights of psiRNA-Con and psiRNA-hH1 groups were 1.66 g ± 0.21 g, 1.48 g ± 0.34 g and 0.59 g ± 0.16 g, respectively. The difference between the normal group and the negative control group was not statistically significant, while the tumor weights of psiRNA-Con and psiRNA-hH1 groups were significantly lower than those of the normal group (t =6.27, P<0.05).

Objective to investigate whether recombinant human acidic fibroblast growth factor (rhaFGF) can promote the proliferation of rabbit skeletal muscle satellite cells. Methods the rabbit skeletal muscle satellite cells were obtained by tissue block culture and density gradient centrifugation. Different concentrations of rhaFGF were added to the cell culture medium, and the control group was set up. The effect of rhaFGF on the proliferation of skeletal muscle satellite cells was evaluated by MTr detection and flow cytometry, and the promoting effect of low molecular weight heparin on this effect was observed. Results rhaFGF can promote the proliferation of skeletal muscle satellite cells. The optimal concentration of rhaFGF is 20~60 μ g/L, and the promoting effect will be reduced when the concentration exceeds 80 μ g/L. Low molecular weight heparin can enhance the above effects of rhaFGF.

Objective to investigate the effect of proteolytic stress induced by environmental toxins on the degeneration and death of dopaminergic neurons. Methods 6-hydroxydopa (6-OHDA), 1-methyl-4-phenylpyridine ion (MPP +) and rotenone were acted on nerve growth factor-induced PC12 cells. The neurotoxic effects of various drugs were detected by MTT assay. The expression and aggregation of α-synuclein and ubiquitinated protein in PC12 cells were observed by confocal immunofluorescence double label staining. The changes of proteolytic enzyme activities in the proteasome of each group of cells were measured by fluorescence spectrophotometer. Results the cell viability of PC12 cells decreased in a concentration-dependent manner after treatment with three neuromutagenic agents. Among them, the cell viability of 6-OHDA 100 μ mol/L, MPP +75 μ mol/L and rotenone 20 nmol/L for 24 h decreased by 52%, 44% and 40% respectively (P<0.01). Confocal microscopy showed that the expression of α-synuclein and ubiquitin in the cytoplasm of cells was up-regulated after treatment with drugs, and protein particles were formed. However, 6-OHDA 100 μ mol/L induced a slight decrease in protease hydrolysis capacity, but the difference was not statistically significant (P>0.05). 6-OHDA 200 μ mol/L induced a further decrease in the activity of three proteases, and the fluorescence index was 7.23 ± 0.58, 79.60 ± 2.73 and 4.23 ± 0.49 respectively (13.90 ± 1.75, 99.30 ± 5.23 and 6.90 ± 0.60 in normal group, P<0.01). Conclusion Neural mutagenic agents can induce the dysfunction of ubiquitin proteasome and the accumulation of α-synuclein and ubiquitinated protein in dopaminergic neurons, and induce abnormal state of proteolytic stress.

A 33-year-old female was admitted to the otolaryngology department on 31 March 2006 due to dry cough, hoarseness and dyspnea for 4 months, which aggravated for nearly 1 month. The patient developed persistent dry cough after catching cold in December 2005, without fever, sputum and dyspnea, and did not improve after antibiotic treatment.

pulmonary artery hypertension (PAH) is a group of pathophysiological syndromes caused by different etiologies and characterized by increased pulmonary artery pressure and pulmonary vascular resistance.

With the increasing use of invasive procedures, the incidence of catheter-related blood infections in the intensive care unit (ICU) is higher, which is not only expensive to treat, but also may lead to death.

The early manifestations of short bowel syndrome (SBS) are mainly diarrhea, dehydration, electrolyte and acid-base balance disorders, which can be aggravated by premature eating.