Volume 87  ·   Issue 03  ·  2007

Objective to investigate the role of CyclinB1 and Cyclin-Dependent kinasel (Cdc2) in the development of endometriosis and their correlation with ovarian hormones. Methods The expression of CyclinB1 and Cdc2 in ectopic endometrium (29 cases) and eutopic endometrium (20 cases) of patients with endometriosis (ectopic group) and endometrium of women with other benign diseases in normal fertility (control group, 30 cases) was detected by RT-PCR, Western blot and immunohistochemistry. Results The expression of CyclinB1 on ectopic endometrium in ectopic group was significantly higher than that in eutopic endometrium and control group (P<0.05). There was no significant difference in the expression of CyclinB1 on eutopic endometrium in ectopic group and control group (P>0.05). There was no significant difference in the expression of Cdc2 in each group (P>0.05). Conclusion The high expression of CyclinB1 may be related to the proliferation of ectopic endometrium in endometriosis.

Objective to investigate the effects of peroxisome proliferative activated receptor (PPAR) γ and its ligands on cytotrophoblast infiltration capacity and its mechanism. Methods The expression of PPAR γ in cytotrophoblast was detected by immunofluorescence cytochemistry, and the effects of different ligands of PPAR γ on the infiltration capacity of primary serum-free cytotrophoblast were detected by Transwell in vitro infiltration system. The regulatory effects of PPAR γ ligands on the expression of matrix metalloproteinase (MMP) -2 and MMP-9 in cytotrophoblast were detected by immunofluorescence confocal technique and fluorescence quantitative PCR. Results PPAR γ protein was mainly localized in cytotrophoblast nucleus, and PPAR γ agonist ligands 15-d-PGJ2 and Troglitazone (TGZ) could inhibit cytotrophoblast infiltration. When the concentrations were 0.1, 1 and 10 μ mol/L, the infiltration indices were 0.7±0.1, 0.6±0.0, 0.4±0.1 and 0.8±0.1, 0.7±0.0 and 0.6±0.0, respectively, which were dose-dependent, and the effect of 15-d-PGJ2 was stronger than that of TGZ, Conclusion PPAR γ plays an important role in the regulation of trophoblast infiltration, and its role may be achieved by regulating the expression of MMP-2 and MMP-9.

Objective to investigate whether there are cognitive-related brain function abnormalities in moderate to severe hypertension patients without heart, brain and kidney complications and cognitive impairment by magnetic resonance functional imaging (fMRI) technique. Methods 50 subjects participated in the experiment. 25 patients with moderate to severe essential hypertension and 25 normal controls were matched according to gender, age and education level. The fMRI stimulation mode was Block Design, and the stimulation materials were presented by auditory. The subjects were asked to listen carefully to nonsensical phrases, while the subjects were asked to listen to true phrases and make psychological judgment on the part of speech (concrete words or abstract words). The fMRI data was processed by SPM 99 software, and the test activation map and negative activation map based on single pixel analysis were obtained. Results The scores of the simple intelligent state examination were 29.3±1.1 in the hypertension group and 29.6±0.5 in the control group, and there was no statistically significant difference between the two groups (P =0.5223). The scores of the anxiety state scale were higher in the patient group (47 ± 5) than in the control group (42 ± 5) (P =0.0356). However, the activity intensity of the brain areas in the patient group was stronger than that in the normal control group. In fMRI experiment 2, the activated and negatively activated brain areas in the two groups were also the same, and the activated brain areas included the superior temporal gyrus and inferior frontal gyrus on both sides; Left angular gyrus; Upper frontal gyrus and premotor area on both sides; The activation intensity and range of the activation in the patient group were greater than that in the control group. Conclusion the patients with moderate to severe hypertension without cardiac, cerebral and renal complications and behavioral cognitive impairment may already have some brain function abnormalities, and fMRI technique may detect these changes early.

Objective to study the anatomy of circular cistern, compare four surgical approaches, and clarify their respective indications and contraindications.Methods 12 cases (24 cerebral hemispheres) of adult skull specimens were fixed with 4% formaldehyde, 4 cases were treated with 4% formaldehyde alone, and 8 cases were intravascularly perfused with colored silica gel.The anatomy and observation of circular cistern were performed with the assistance of surgical microscope.The four surgical approaches to circular cistern were compared, and the structures restricting each approach.Results the circular cistern extends from the posterior edge of the foot cistern to the lateral edge of the midbrain tetrass, and surrounds the lateral surface of the upper brainstem, which mainly contains the P2 segment of posterior cerebral artery, superior cerebellar artery, anterior choroidal artery, basilar vein and trochlear nerve.The subtemporal approach is suitable for the lesions of the inferior part of circular cistern, and the exposure of Labbé vein and parahippocampal gyrus affects the exposure of this approach; Trans-temporal-trans-choroidal approach is suitable for non-dominant hemispheric circular cistern lesions. This approach requires temporal cortical incision, and the anterior Labbé vein will limit the cortical incision; The transinsular-transchoroidal fissure approach is suitable for anterior and superior peripheral lesions, but it can damage the temporal trunk. The longer the inferior peripheral sulcus, the shorter the incision and the smaller the damage; The occipital transtentorial approach is suitable for posterior and inferior circular cistern lesions, and the parahippocampal gyrus is also the main structure influencing this approach.Conclusion The accurate location of the lesion must be carefully considered in the selection of surgical approach for circular cistern lesions. The relationship between Labbé vein, parahippocampal gyrus and the lesion is the key to determine the surgical approach.

Objective to investigate the feasibility of reverse transcription polymerase chain reaction (RT-PCR) in the detection of lymph node micrometastasis in non-small cell lung carcinoma. Methods Each lymph node was divided into two halves and one half of the lymph node was examined by HE staining. The other half lymph nodes were mixed by region and used for RT-PCR. If one lymph node was confirmed by HE staining, the other lymph nodes in the same region were no longer subjected to RT-PCR. Results (1) A total of 195 lymph nodes of 25 patients with lung cancer were examined by HE staining, and 9 of 30 lymph nodes were found to have dominant metastasis, and none of the lymph nodes were detected with micrometastasis. (2) Among 39 groups of mixed lymph nodes with HE staining negative, 11 groups were positive for RT-PCR. (3) Among 16 patients with routine pathological PN0 stage, 6 cases had micrometastasis in hilar lymph nodes; The difference was statistically significant (χ 2=54.063, P =0.0043). Conclusion (1) HE staining pathology can accurately detect the dominant lymph node metastasis in NSCLC lymph nodes, but it is not easy to detect the occult micrometastasis. (2) RT-PCR can improve the detection rate of NSCLC lymph node micrometastasis, and can re-stage some stage Ⅰ and Ⅱ patients.

Objective to investigate the relationship between human epidermal growth factor receptor-2 (HER-2) gene and lymphangiogenesis in breast cancer patients and its effect on prognosis, in order to investigate the molecular mechanism of poor prognosis in patients with HER-2 overexpression. Methods (1) Herceptin blocked the function of HER-2 protein in breast cancer SKBR-3 and MDA-MB-231 cell lines in vitro, and the expression of vascular endothelial growth factor-C (VEGF-C) mRNA and protein before and after blockade was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. (2) The expression of VEGF-C mRNA in 67 cases of invasive ductal carcinoma of breast was detected by real-time qRT-PCR. (3) The expression of HER-2 and VEGF-C protein in paraffin-embedded tissues of 160 cases of breast cancer were detected by immunohistochemistry, and the lymphatic vessel density (LVD) was counted by podoplanin antibody, and the lymphatic vessel invasion (LVI) was observed. Results (1) The expression of HER-2 was high in SKBR-3 after herceptin blockade . (2) The expression of VEGF-C mRNA in lymph node metastasis group (1.6±0.5) was higher than that in non-metastasis group (1.1±0.5) (t =2.196, P<0.05). Logistic regression model showed that HER-2, LVD and LVI were important factors affecting lymph node metastasis (all P<0.05). (3) Kaplan-Meier survival analysis showed that HER-2 protein overexpression, VEGF-C protein overexpression, high LVD and LVI were all related to low disease-free survival rate of patients (all P<0.05). Cox risk ratio model analysis showed that HER-2 and LVI were independent prognostic factors in breast cancer (all P<0.05). Conclusion HER-2 overexpression in breast cancer upregulated VEGF-C expression, promoted lymphangiogenesis and tumor lymph node metastasis were important molecular mechanisms for poor prognosis of patients. VEGF-C expression, LVD and LVI were new important indicators to evaluate the prognosis of breast cancer patients.

Objective to analyze the expression of Fas, Fas ligand (FasL), Fas associated death domain protein (FADD), aspartate specific cysteine protease 8 (caspase8) and mutant P53 protein in esophageal squamous cell carcinoma, and to investigate the relationship between these five apoptosis-related proteins and the occurrence and development of esophageal squamous cell carcinoma. Methods A tissue microarray containing 116 esophageal squamous cell carcinoma patients and corresponding normal esophageal squamous cell tissues was constructed. The expression of Fas, FasL, FADD, caspase8 and mutant P53 protein in squamous cell carcinoma and squamous cell tissues was detected by immunohistochemistry, and the correlation between the expression differences and clinicopathological features and prognosis was analyzed. Results the expression of Fas in squamous cell carcinoma was lower than that in normal squamous cell carcinoma (positive rates were 41.4% and 95.7%, respectively, P<0.001). The expression of FasL and FADD in squamous cell carcinoma (positive rates were 76.7% and 50%, respectively) was up-regulated compared with that in normal squamous cell carcinoma (positive rates were 39.7% and 7.8%, respectively) (both P<0.01). The expression was diffuse in squamous cell carcinoma tissues (positive rate 37.1%). The disease-free survival rate of FADD positive patients was lower (P =0.0028); However, the expression of Fas, FasL, caspase8 and mutant P53 protein was not significantly related to the disease-free survival rate (all P>0.05). Conclusion Fas, FasL, FADD, caspase8 and mutant P53 protein are closely related to the occurrence and development of esophageal squamous cell carcinoma, and are important markers of the occurrence and development of esophageal squamous cell carcinoma. Among them, FADD can be used as a prognostic marker of esophageal squamous cell carcinoma.

Objective to investigate the presence of angiogenic mimicry (VM) in patients with primary gallbladder carcinoma and its clinical significance. Methods The pathological specimens and related clinicopathological parameters and prognostic indexes of 74 patients with primary gallbladder carcinoma diagnosed by surgical resection and pathology were collected from January 1994 to December 2000. HE staining, CD31 and PAS double staining were used to observe the presence of VM in patients with gallbladder carcinoma, and univariate analysis, Kaplan-Meier survival comparison and Cox risk model were performed. Results VM was found in 13.5% (10/74) of 74 patients with gallbladder carcinoma. But it was related to tissue type (χ 2=10.241, P =0.017), liver metastasis (χ 2=11.904, P =0.001) and survival time (χ 2=5.7221, P =0.0168). Cox analysis showed that VM (F =24.0508, P =0.0001, relative risk =11.868) was the same independent risk factor affecting the prognosis of patients with gallbladder carcinoma as depth of invasion, lymph node metastasis, liver metastasis and surgical method (palliative surgery).

Objective to investigate the feasibility, effect and mechanism of using controlled tube structure scaffold to inoculate osteoblasts and parallel rotary dynamic three-dimensional culture as a new method for constructing segmental tissue engineering bone in vitro.Methods A controlled tube structure self-curing calcium phosphate (CPC) scaffold was designed and prepared, and a new rotary bioreactor was designed and developed.After seeding the scaffold, rabbit skull osteoblasts were isolated and expanded, respectively, and then cultured in rotation and static culture for 7, 14 and 21 days, respectively.After that, tetramethylazolazole blue (MTT) colorimetry,daily glucose consumption,cell alkaline phosphatase (ALP) activity,scanning electron microscopy (SEM) observation,X-ray energy spectroscopy (EDX) analysis were performed. Results The proliferation, metabolism and osteogenic differentiation of cells in rotary culture group were significantly better than those in static culture (F =144.187,F =491.603 and F =34.913, all P<0.01 and P<0.01); Conclusion CPC scaffold with controlled tubular structure combined with rotating dynamic three-dimensional culture can effectively promote the proliferation, metabolism, differentiation and rational distribution of osteoblasts in scaffold, which can be a new method for constructing segmental tissue engineering bone in vitro.

Objective to investigate the expression of nuclear factor-κ B (NF-κ B) in bronchoalveolar lavage cells of mice with pulmonary interstitial fibrosis induced by bleomycin (BLM) and its effect on interleukin-4 (IL-4), and the intervention effect of its antisense oligonucleotide (ASON) on pulmonary interstitial fibrosis. Methods C57B1/6 mice were randomly divided into 4 groups. The control group and BLM group were injected with normal saline through tail vein, the ASON group was injected with p65 ASON, and the SON group was injected with p65 cis-sense oligonucleotide (SON). After 6 hours, the BLM group, ASON group and SON group were injected with bleomycin-A5 (BLM-A5) intratracheally, and the control group was injected with normal saline. The mice in the control group were sacrificed at 0.5, 1, 14 days after tracheal injection, and the other groups were injected with bronchoalveolar lavage at 0.5, 1, 3, 7, 14, 28 days after tracheal injection. A small amount of collagen fibers were observed in lung interstitium. (2) The content of hydroxyproline in lung tissue of BLM group and SON group was significantly higher than that of control group and ASON group was significantly lower than that of BLM group and SON group. (3) The expression of p65 in bronchoalveolar lavage cells of BLM group and SON group was significantly higher than that of control group and ASON group. (4) The expression of IL-4 in bronchoalveolar lavage cells of BLM group and SON group was significantly higher than that of control group and ASON group. (5) The content of IL-4 in bronchoalveolar lavage fluid of BLM group and SON group was significantly higher than that of control group and ASON group. (6) The expression of p65 in bronchoalveolar lavage cells of mice in BLM group and ASON group was positively correlated with IL-4 (r =0.890, P<0.05; r =0.909, P<0.05). Conclusion During the pathological process of pulmonary interstitial fibrosis induced by BLM, the expression of NF-κ B in bronchoalveolar lavage cells was significantly enhanced, and may form pulmonary interstitial fibrosis through IL-4 pathway. NF-κ B ASON can significantly inhibit the activation of NF-κ B

Objective to study the induction of osteoblast differentiation of rat bone marrow-derived mesenchymal stem cells (MSCs) by immunosuppressant tacrolimus (FK506), and the in vivo osteogenic effect of MSCs combined with porous β-calcium triphosphate (β-TCP). Methods Primary MSCs were randomly divided into groups according to the experimental objective.L-ascorbic acid-2-phosphate (AsAp) and β-glycerophosphate (β-GP) were added to the growth medium of control group; In the FK506 treatment group, 50 nmol/L FK506 was added to the control group. The experiment was divided into two parts in vitro and in vivo successively. In vitro, the activity of alkaline phosphatase (APase), calcium content, scanning electron microscope and osteocalcin mRNA Northern blot were analyzed on the 4th, 8th, 12th and 16th days of culture respectively. In vivo, cells were loaded into porous β-TCP cubes to form MSCs/β-TCP complexes, which were implanted subcutaneously on the back of rats after continued osteogenic induction culture for 2 weeks, and the complexes were removed for histological examination after 4 and 8 weeks. Results the activity of alkaline phosphatase, calcium content and osteocalcin mRNA in the FK506 treatment group were significantly higher than those in the control group (all P<0.05); Conclusion Immunosuppressant FK506 can significantly promote osteoblast differentiation of rat bone marrow-derived mesenchymal stem cells (MSCs) in vitro and continuous osteogenesis in animals, and it suggests that FK506 can be used as a new osteogenic growth factor with potential clinical application value.

Objective to study the therapeutic effect and mechanism of bone marrow mesenchymal stem cells (BMSC) on ischemic stroke in rats. Methods BMSC was isolated and cultured by density gradient centrifugation. A rat model of middle cerebral artery embolism-ischemia reperfusion for 2 h was established. Sixty cerebral ischemia model rats who met the criteria were randomly divided into a treatment group (48 rats in BMSC transplantation group) and a control group (12 rats in D-Hanks solution injection group) according to the nerve injury score. After 24 h, hBMSC suspension and D-Hanks solution were injected into the cortex of the marginal area of cerebral ischemia, respectively. Cycle testing of the adhesive removal experiment, rotary rod experiment and Morris water maze experiment was carried out on the 4th day after transplantation until the 32nd day; In the treatment group, the rats were sacrificed in batches at 1, 2, 3 and 4 weeks after transplantation, respectively. BMSC were dynamically tracked by immunofluorescence technique, and the cell migration and differentiation were detected. Results A large number of cells survived and migrated to the lesion at 1 and 2 weeks after transplantation, the number of surviving cells decreased significantly at 3 weeks, and very few surviving cells at 4 weeks; hBMSC did not clearly express microtubule-associated protein (MAP) 2, glial fibrillar acidic protein (GFAP) and galactocerebroside (GalC) in rat brain. The mean escape time of water maze at 1 week after treatment was (69±10) s in the treatment group and (120±0) s in the control group, P<0.05. The difference between the two groups was not statistically significant at 4 weeks (P>0.05). The mean incubation period of rotating rod experiment (10 r/min) at 10 days after treatment was (167±18) s in the treatment group and (37±19) s in the control group, P<0.05. The difference was still statistically significant at the end of the experiment. The mean incubation period of adhesive removal experiment at 13 days after treatment was (33±8) s in the treatment group and (84±13) s in the control group, P<0.05. The difference was still statistically significant at the end of the experiment. Conclusion BMSC intracerebral transplantation can significantly improve the recovery of neurological function in rats with cerebral ischemia; The transplanted cells can migrate to the periphery of the rat brain, but the survival of the cells has a certain time limit; BMSCs did not clearly differentiate into transcellular cells in the rat brain.

The patient was a 46-year-old woman who was pregnant and delivered 2 days. Due to heavy menstrual flow for 1 year, massive vaginal bleeding for 2 days accompanied by dizziness and fatigue, she was hospitalized in Sir Run Run Shaw Hospital affiliated to Zhejiang University on June 2, 2005. The patient developed heavy menstrual flow without obvious trigger 1 year ago, with a few blood clots, prolonged menstrual period to 10 days/time, and no lower abdominal pain during menstruation. When the patient had menstruation 2 days before the hospital, the menstrual flow increased significantly compared with before, with a large number of dark purple blood clots, and the patient developed dizziness and fatigue. Physical examination: blood pressure: 110/70 mm Hg (1 mm Hg =0.133 kPa), heart rate 100 beats/min. Anemia appearance. Gynecological examination: no abnormality in vulva, massive blood accumulation in vagina, severe cervical erosion, dark red blood accumulation in cervical canal, the size of uterine body as more than 3 months of pregnancy, medium mass, movable, no obvious tenderness, and no obvious abnormality in double adnexal area.

In the 1970s, researchers discovered that extracts from other tissues other than thymus tissue also have a similar effect to thymopoietin in promoting T lymphocyte differentiation. Based on this discovery, in 1975, Gideon Goldstein and Margrit Scheid of New York University School of Medicine isolated and purified a polypeptide involved in the differentiation of T and B lymphocytes with a molecular weight of about 8500 from bovine thymus. It was proved that it exists widely in all kinds of eukaryotic cells and has high similarity in structure, function and immunology among various species. Goldstein et al. concluded that this genetic conservation is a manifestation of its important role in life activities, and named it ubiquitous immunopoietic polypepfide (UBIP), which was later called "ubiquitin".

Estrogen in premenopausal women is mainly secreted by the ovaries. Aromatase, a member of the family of enzyme complexes containing cytochrome P450, acts on the rate-limiting step of estrogen production, i.e., catalyzes the conversion of androstenedione or testosterone to estrogen. This process is controlled by follicle-stimulating hormone (FSH) and luteinizing hormone (LH) secreted by the pituitary gland.

Whether hormone replacement therapy (HT) is safe to relieve perimenopausal symptoms and to treat postmenopausal osteoporosis, and whether it will increase the occurrence of cardiovascular and cerebrovascular diseases and related tumors, has been a hot research topic.Early epidemiological studies mainly focused on the relationship between estrogen therapy (ET) and stroke.The latest studies mainly focused on the relationship between estrogen and progesterone combination therapy (EPT) and risk factors of cerebrovascular diseases, suggesting that progesterone may increase the risk of cerebrovascular diseases.

The patient was a 13-year-old male from Fuyang City, Zhejiang Province. He came to our hospital on February 8, 2006 for "repeated abdominal pain, diarrhea, progressive weight loss for half a year, and the abdominal pain worsened for two days". He was admitted to the hospital with the intention of "abdominal pain to be examined". The patient had repeated abdominal pain in the past six months, presenting paroxysmal pain in the upper abdomen, which was tolerable and had nothing to do with eating. At the same time, he was accompanied by black paste stool, 5~6 times a day, no blood and mucus, no tenesmus, no fever, no vomiting, and gradually appeared abdominal distension and progressive weight loss. The patient had swallowed the snake gallbladder of the five-step snake 4 months ago. He went to the local hospital for abdominal pain 1 month ago and was diagnosed with "abdominal lymphadenitis?" and was given anti-inflammatory treatment. Because the disease did not improve, he was hospitalized in our hospital for 1 week on January 18, 2006 and then discharged.

Organized and compiled by the National Pharmacopoeia Commission<Instructions for Clinical Use of Pharmacopoeia of the People's Republic of China (2005 Edition) (abbreviated as the book) was officially published by People's Medical Publishing House in December 2005. The book is the 2005 edition<Pharmacopoeia of the People's Republic of China<Chinese Pharmacopoeia (Chinese Pharmacopoeia) is one of the important supporting books, and it is also a clinical reference book mainly for medical staff and pharmaceutical enterprises in China.