BACKGROUND:Annulus puncture method is commonly reported in the establishment of intervertebral disc degeneration (IDD) models, but there is a lack of comparative studies concerning complete MRI, pathologyical and extracellular matrix changes. Thereafter, a further exploration is imperative. OBJECTIVE:To investigate the feasibility of establishing IDD models using annulus puncture. METHODS:Thirty New Zealand white rabbits were randomly divided into two groups (n=15 per group). In experimental group, through extraperitoneal approach, the L1-2, L2-3, and L3-4 intervertebral space were exposed, and then stabbed by 16-gauge hypodermic needles;while rabbits in sham operated group were only subjected to making and incision but not to puncture. Five rabbits in each group were randomly selected for MRI examination at 4, 8 and 16 weeks after modeling, and were sacrificed through air embolism to obtain the nucleus pulposus. Histological changes were observed by hematoxylin-eosin staining, and the level of proteoglycan was detected by Safranin O staining. RESULTS AND CONCLUSION:In the experimental group, T2-weighted signal in the disc was decreased in a time-dependent manner;an obvious reduction was found at 4 weeks after modeling, and reached the peak at 16 weeks. In contrast, the sham operated group showed no significant changes at each time point. Hematoxylin-eosin staining showed that in the experimental group, cartilage-like cells decreased, while fibroblast-like cells in the disc increased in number and proportion, and extracellular fibers were in disorder;all above changes became obvious at 4 weeks after surgery and the most obvious at 16 weeks, but there were no significant changes in the sham operated group. Safranin O staining revealed that the color of the tissues under puncture became lighter, and this variation became overt at 4 weeks after surgery and the most obvious at 16 weeks. In conclusion, annulus puncture is a reliable method to make IDD models.

Objective To compare the effect of the different operation for mammary duct fistula (MDF). Method The clinical data of 85 cases of MDF were analysed retrospectively. Continuous section examinations between the multiple fistula was performed in 3 cases undergoing breastectomy.Results Among the 85 cases with 146 MDF, the opening located within 5cm around areola in 129 cases. MDF incision was performed on 38 cases, 34 cases cured, 4 cases recurred;MDF resection was performed in 44 cases, all the 44 cases were cured;simple mastectomy was performed on 3 cases of extensive breast inflammation,all the 3 cases cured. Conclusions Most of MDF occurs in main mammary duct. Most of the opening of MDF are around the areola. MDF incision or resection is the first choice of operation.

AIM: To transfer 4 full-length WT1 isoforms cDNA into the leukemia cell line NB_4 so as to provide a cell model for studying the WT-1 gene function. METHODS: The eukaryotic expression recombinant vectors for WT1 isoforms (pCB6+/WT1) were introduced into the leukemia cell line NB_4 by electroporation. The positive cell clones were screened by G418 culture. The integration of WT1 gene isoforms in NB_4 cells as confirmed by PCR. The mRNA and protein of WT1 were detected by RT-PCR and Western blotting. RESULTS: WT1 gene isoforms were successfully transferred into NB_4 cells. WT1 mRNA and protein expression in the G418-selected cells increased remarkably compared with the control. CONCLUSION: WT1 gene isoforms were effectively transferred into NB_4 cells by electroporation and stably expressed in the transfected cells.

Objective To investigate the effect of Shanzha Xiaozhi capslue on intima-media thickness(IMT) and plaques score of carotid artery in patients with non-acute phase coronary heart disease and traditional Chinese medicine(TCM)syndrome of phlegm and blood stasis. Methods A prospective study was conducted to carry out a research on 102 patients with non-acute phase coronary heart disease and TCM syndrome of phlegm and blood stasis. They were randomly divided into two groups:a control group(50 cases)treated with conventional western medicine alone and a observation group(52 cases)which was treated by both conventional western medicine and Shanzha Xiaozhi capslue(the main TCM ingredients:Shanzha,Dahuang)0.7 g,3 times a day,the therapeutic course being 6 months in both groups. The cardiocerebral vascular incidences of the two groups were observed,in the mean time,the carotid artery IMT and plagues score were registered,the scores of phlegm stagnation syndrome and blood-stasis syndrome were measured,adverse reaction was observed and compared in both groups before and after treatment. Results After treatment,the occurrence of major adverse acute cardiovascular and cerebrovascular events in observation group were reduced significantly compared with those in the control group(5.77%vs. 20.00%,P<0.05). The IMT and plaques score of carotid artery and the scores of phlegm stagnation syndrome and blood-stasis syndrome were all decreased obviously compared to those before treatment,and the effect was more remarkable in the treatment group〔IMT of carotid artery(mm):0.80±0.13 vs. 0.95±0.12,the plaques score:1.35±0.65 vs. 1.75±0.88, phlegm syndrome score:20.98±6.42 vs. 35.55±9.22,blood-stasis syndrome score:23.23±5.12 vs. 28.95±6.38, P<0.05 or P<0.01〕. Abdominal pain appeared in 2 patients of observation group without other adverse reactions. Conclusion Shanzha Xiaozhi capsule can stabilize atherosclerotic plaque and reduce the occurrence of acute adverse cardiocerebral vascular events in patients with non-acute phase coronary heart disease and phlegm and blood stasis syndrome possibly by removing phlegm and blood stasis to decrease the IMT and plaques score of carotid artery.

Objective To compare early-stage clinical features and hypothalamic-pituitary-adrenal (HPA) axis neuroendocrine function in first-episode and untreated major depressive disorders with suicide and without suicide. Methods Untreated patients who had a diagnosis as major depression according to ICD-10 were allocated to suicide with major depressive disorder group or non-suicide with major depressive disorder group according to whether the pa?tients had suicidal ideation or behaviours. Patients were assessed before treatment using the 17-item Hamilton Depres?sion Scale (HAMD-17), the Hamilton Anxiety Scale (HAMA), the Yale-Brown Obsessive Compulsive Scale (YBOCS) and the suicide assessment scale. The cortisol (CORT) and adrenocorticotropic hormone (ACTH) levels in serum were measured using a chemiluminescence immunoassay. Results There was no significant difference in demographic data be?tween the two groups (P>0.05). In early-stage clinical features, there were significant differences in changes of sexual ac? tivity and feeling of despair (P<0.05). HAMD total scores were significantly higher in the suicide group (P<0.01), whereas HAMA total scores and Y-BOCS total scores were not significantly different between the two groups (P>0.05). There were no significant differences in CORT or ACTH levels between two groups (P>0.05). There were no significant correlation of suicidal scores with early-stage clinical features, symptom scales scores, CORT or ACTH levels (P>0.05). Conclusion De?creased sexual desire and feeling of despair are more severe in depressed patients with suicide than those without. There is no correlation of early-stage clinical features or HPA axis functions with suicide in patients with depression.

AIM: To investigate the protective effect of the bone marrow cells transfected with human multidrug resistance gene (MDR1) on the reconstruction of murine hematopoietic function.METHODS: The mononuclear cells of the bone marrow from donors, BALB/C mice, treated with 5-Fu previously, were isolated and transfected with human multidrug resistance gene in vitro , then transplanted to the tertiary recipients. After lethal irradiation(8.5 Gy) and bone marrow transplantation, the recipients were selected with Taxol 7 mg/kg intraperitoneal injection, VCR 5 mg/kg or DNA 5 mg/kg intravenous injection. The survival rate and blood pictures of mice as well as the integration and expression of target gene MDR1 were studied. RESULTS: The lethal irradiated murine hematopoietic function could be reconstructed and protected from toxicity of high doses Taxol, VCR and DNR selection after reinfusing the hematopoietic progenitor cells containing human multidrug resistance gene (MDR1). The survival rate and survival time of experimental mice were higher than that in the control group. The integration and expression of MDR1 gene in recipients were confirmed by PCR, RT-PCR and FCM. CONCLUSION: The integration and expression of human multidrug resistance gene in recipients may play an important role in the reconstruction and protection of murine hematopoietic function.

AIM: To explore the effects of cholestoral and mevalonate synthesis inhibitor lovastatin (LOV) on the proliferation of NB4 cells and elucidate the mechanisms. METHODS: Cell proliferation was analyzed by MTT assay;the expression of H, K, N- ras oncogenes in NB4 cells at different time point after LOV treatment were determined by semi-quantitative RT-PCR. Both total p21 Ras protein and p21 Ras protein on the cellular membrane were examined by flow cytometry. RESULTS:①LOV inhibited the proliferation of NB4 cells. ②All three kinds of ras were expressed in NB4 cells. ③LOV caused no increase in H, K, N- ras mRNA level. Amount of total p21 Ras protein did not change as the time varied. Concomitantly,p21 Ras protein localized on the cellular membrane decreased. CONCLUSION:LOV inhibits the proliferation of NB4 cells. Targeting HMG-CoA reductase, LOV blocks the isoprenylation of p21 Ras protein which affects its anchorage on the cellular membrane. No change in the H, K, N- ras mRNA and total p21 Ras protein expression is detected.

OBJECTIVETo present a special case with the karyotype and molecular marker of acute myeloid leukemia (AML)-M2 who was induced to complete remission by all-trans retinoic acid (ATRA) alone.

METHODSA recently hospitalized young female patient with acute leukemia was initially diagnosed as M3 subtype based on morphological French-American-British (FAB) classification. Karyotype analysis using standard G and R banding techniques and RT-PCR were applied to further define the diagnosis. After primarily cultured bone marrow cells from the iliac aspiration were tested for in vitro induced differentiation, the patient was treated with oral all-trans retinoic acid alone, 60 mg per day until complete remission was achieved. Peripheral blood and bone marrow changes were monitored over the whole treatment course.

RESULTSThe characteristic chromosomal aberration for M3, the t(15;17) reciprocal translocation, was not found while a t(8;21) translocation was verified. Furthermore, an amplified product of the AML-1/ETO fusion gene instead of the PML/RAR alpha fusion gene was detected by RT-PCR and the diagnosis was corrected from M3 to M2. Primary cultured bone marrow cells can be fully induced to terminal differentiation after 4 days exposure to ATRA. A hematological complete remission was achieved after 40 days treatment with ATRA as a single therapeutic agent, suggesting an alternative pathway mediating ATRA-induced myeloid differentiation.

CONCLUSIONA leukemia patient with a subtype other than M3, such as M2 in this case, may also be induced to complete remission by the mechanism of ATRA-induced terminal differentiation. This implies that there may be a pathway other than PML/RAR alpha fusion gene product which mediates ATRA-induced myeloid maturation in leukemia cells.

Adolescent ; Antineoplastic Agents ; therapeutic use ; Core Binding Factor Alpha 2 Subunit ; Female ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; Neoplasm Proteins ; analysis ; genetics ; Oncogene Proteins, Fusion ; analysis ; genetics ; physiology ; RUNX1 Translocation Partner 1 Protein ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; physiology ; Tretinoin ; therapeutic use

Retroviral vectors are wildly used as vehicles for gene transfer into hematopoietic cells based on its potency for efficient gene delivery and integration of transgene in host genome. The development of better transduction protocols depends on gene markers that allow a rapid detection and effective selection of genetically transduced cells. In this study, the enhanced green fluorescent protein (EGFP), a gene that is optimized for detection and expression in mammalian cells, was firstly amplified and cloned by high-fidelity PCR. The vector LGSN carrying EGFP gene was then constructed and the retroviral producer cell lines that yield high titers of LGSN vector in supernatants were developed by liposome-mediated transfection in combination with cross infection. Both GP + envAm12 murine fibroblasts and K562 leukemic cells transduced with EGFP virus demonstrated a stable green fluorescence signal readily detectable by flow cytometry or fluorescence microscopy in up to 97% and 86% of examined cells, respectively. The integration of LGSN provirus in transduced cells was confirmed by PCR analysis. These results indicate EGFP is a suitable reporter molecule for gene transfer and expression in hematopoietic cells. Therefore, the bright and long-term expression of EGFP in living cells will advance the study of gene therapy in vitro and in vivo, particularly for human applications.

Objective:To explore the clinical value of peripheral remnant lipoproteins (RLP), low density lipoprotein cholesterol particle (LDL-P) and sdLDL particle (sdLDL-P) measurement in the diagnosis of carotid plaque, so as to provide practical basis for the accurate diagnosis of carotid plaque and the control of carotid plaque related cardiovascular and cerebrovascular diseases.Methods:People who underwent carotid plaque ultrasound examination in Xingtai Third Hospital , from January 2020 to June 2021 were selected as the research object. According to the ultrasound results, they were divided into carotid plaque group ( n=146) and control group without carotid plaque ( n=149). The fasting RLP, LDL-P and sdLDL-P of the two groups were measured by vertical auto profile (VAP) centrifugal separation phase, and the fasting TG and LDL-C were detected by routine mixed phase method. The indexes were compared between the two groups and the true positive rate, true negative rate, false positive rate and false negative rate of the diagnosis of carotid plaque were analyzed. The receiver operating characteristic curve of each test index was drawn, and AUC was used to evaluate the clinical diagnostic value of each test index for carotid plaque. Results:The levels of RLP, LDL-P and sdLDL-P in carotid plaque group were significantly higher than those in non-carotid plaque group ([1.07±0.36] mmol/L vs [0.59±0.17] mmol/L,[1 300±370] nmol/L vs [781±215] nmol/L,[435±139] nmol/L vs [156±59] nmol/L, all P<0.01). The true positive rate (78.08% [114/146],81.51% [119/146]) and true negative rate (84.56% [126/149], 86.58%[129/149]) of serum RLP and LDL-P for the diagnosis of carotid plaque were significantly higher than TG (58.90%[86/146], 43.62%[65/149]) and LDL-C (59.59% [87/146], 46.98% [70/149]), and the false positive rate (15.44% [23/149], 13.42% [20/149]) and false negative rate (21.92% [32/146], 18.49% [27/146]) were significantly lower than TG (56.38% [84/149], 41.10% [60/146]) and LDL-C (53.02% [79/149], 40.41% [59/146], all P<0.01). The AUC of the ROC curve of RLP (0.890), LDL-P (0.902) and sdLDL-P (0.973) for the diagnosis of carotid plaque was higher than TG (0.682) and LDL-C (0.712). The AUC of ROC curve of the RLP combined with sdLDL-P (0.977) for the diagnosis of carotid plaque was higher than the RLP and sdLDL-P (all P<0.01). Conclusion:The serum RLP, LDL-P and sdLDL-P can be used as indicators of carotid plaque, and their clinical diagnostic value are superior to TG and LDL-C; the combined diagnostic effect of lipoprotein subclass is better than that of single index alone.