Objective: To explore the effect of psychotherapy in treating middle school students with IAD (internet addiction disorder). Methods: 23 cases of middle school students with IAD had been treated by cognitive therapy or behavior therapy. The IAD scale, SCL-90 scale and SDS scale were used to evaluate the effect before and after therapy. Results: After therapy, the total scores and most dimensions scores in IAD scale, SCL-90 scale and SDS scale decreased significantly than that before therapy. Conclusions: Psychotherapy especially cognitive therapy and behavior therapy is an effective in treating middle school students with IAD.

Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.

Background Most animal models of experimental autoimmune uveitis (EAU) are single attacked procedure,with a different feature from the natural course of human recurrent autoimmune uveitis.So establishing a recurrent EAU model is necessary for the clinical study on EAU.Objective This study was to establish the recurrent EAU model in rat and investigate the ocular inflammation and pathological manifestation and interleukin-17 (IL-17)expression in the eye.Methods T cells isolated from the spleen and draining lymph nodes of Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 peptide fragments (R16) 10 days earlier were re-stimulated with R16 in vitro and injected into naive syngeneic rats to establish the recurrent EAU models,and the normal Lewis rats were used as controls.The eyes of model rats were then examined daily for clinical signs of uveitis by slit-lamp biomicroscopy and scored Caspi's criteria.The rats were sacrificed 1 month,2,3months after injection respectively,and the retinal sections were prepared for the pathological examination by hemotoxylin & eosin staining.Immunohistochemistry was performed to detect the expression of IL-17 in the retina.Results Adoptive transfer of R16-specific T cells to Lewis rats induced recurrent uveitis.The inflammatory scores on the fourth day,the sixth day,and the inflammatory response disappeared on the tenth day after injection.However,the inflammatory reaction occurred repeatedly 4 or 5 times in the 2-month duration after that,and the right and left eyes of a single recipient showed a different pattern of relapse,and the clinical manifestations of EAU was similar to the natural course to those of human autoimmune uveitis.In the retinal specimens of 1-,2-and 3-month group,the number of inflammatory cells was gradually decreased as the time lapse.Compared with the normal group,the thicknesses of the entire retina,outer nuclear layer and inner nuclear layer decreased with a significant difference among the 4 groups (F=20.46,288.40,4.43,all P=0.00).The number of RGCs in the normal group,1-,2-and 3-month group was 231.27 ± 15.36,225.36 ± 17.79,132.18 ±9.39 and 67.45 ± 11.90,respectively,showing a significant difference among them (F=68.94,P=0.00).Immunohistochemistry showed that the scores of the IL-17 expression in the rat retina were 0.64 ± 0.17,1.92 ± 0.19,1.17 ± 0.23 and 0.83 ± 0.23,showing statistically significant difference (F=64.10,P=0.00).Conclusions The stimulation of R16-specific T cells can induce recurrent EAU in Lewis rat.Th17 is involved in the disease course.

OBJECTIVETo investigate the growth and development of nerve growth factor (NGF)-positive neurons in the cerebellum of midanaphase human fetus.

METHODSThe expression of the NGF-positive neurons in the cerebrum of human fetus was observed by immunohistochemical methods, and the integral absorbance (IA) was detected.

RESULTSBy the 3rd to 4th month of gestation, neurons was seen in the ependymal, central, and marginal plate of cerebellum; the nucleus was oval and the neurons had short and small processes. By the 5th to 7th month of gestation, the number of NGF-positive neurons increased, the expressions enhanced, the nucleus was round-, oval-, or fusiform-shaped, the neurons grew larger in size, and the Purkinje cells showed NGF-positive expression. By the 8th to 10th month of gestation, the NGF-positive expression was enhanced with deeper dying, the body of Purkinje cells grew larger gradually, and the number of NGF-positive neurons in the granular cell layer and molecular layer increased. IA of the cerebellar cortical neurons of the 3rd, 4th, 5th, 6th, 7th, and 8th month of gestation showed an increasing trend, and significant difference was observed (P < 0. 05).

CONCLUSIONNGF-positive neurons in the cerebellum play an important role for differentiation, proliferation, migration, and growth of neurons in the cerebellum.

Cerebellum ; cytology ; metabolism ; Fetus ; cytology ; metabolism ; Humans ; Nerve Growth Factor ; metabolism ; Neurons ; cytology ; metabolism ; Purkinje Cells ; metabolism

OBJECTIVETo study the expression of matrix metalloproteinase (MMP)-2, MMP-9 and vascular endothelial growth factor (VEGF)-C in breast cancer and their role in lymph node metastasis.

METHODSImmunohistochemical staining was used to detect the expression of VEGF-C, MMP-2, MMP-9 and LYVE-1 in 84 cases of breast cancer, including 52 cases with and 32 cases without lymph node metastases. The recombinant vector (pSIREN-VEGF-C) was transfected into human breast cancer cell MCF-7 by liposome, and the RNA expression of MMP-2, MMP-9 and VEGF-C in MCF-7 cells after transfection was detected by PCR.

RESULTSThe expressions of MMP-2, MMP-9 and VEGF-C were 98.1% (51/52), 88.5% (46/52), and 94.2% (49/52) respectively for the metastatic group, and 75.0% (24/32), 53.1% (17/32), and 65.6% (21/32) respectively for the non metastatic group, and there was significant difference between these groups (P < 0.05). The lymphatic vessel density between these two groups was also significantly different (P < 0.05). Increased expression of MMP-2, MMP-9 and VEGF-C was also associated with increased number of lymphatic vessels had also increased (P < 0.05). The expression of MMP-2, MMP-9 and VEGF-C in MCF-7 cells after gene transfection decreased significantly (P < 0.05).

CONCLUSIONMMP-2 and MMP-9 in conjunction with VEGF-C, promote lymphangiogenesis and lymph node metastasis of breast cancer.

Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; surgery ; Carcinoma, Medullary ; genetics ; metabolism ; pathology ; surgery ; Cell Line, Tumor ; Female ; Humans ; Lymph Nodes ; metabolism ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Middle Aged ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; Vesicular Transport Proteins ; metabolism

OBJECTIVETo investigate the appearance and clinical value of MRI in the diagnosis of embryonal rhabdomyosarcoma in nasal cavity and paranasal sinuses.

METHODSThe clinical, pathological and MRI findings of five patients with pathology proved embryonal rhabdomyosarcoma in nasal cavity and paranasal sinuses were retrospectively analyzed.

RESULTSThe major clinical symptoms included nasal obstruction (4 cases), exophthalmus (4 cases), decreased eyesight (2 cases), epistaxis (1 case), decreased olfactory sensation (1 case) and restriction of eyeball movement (1 case). All 5 cases involved multi-location. Among them, 4 cases mainly located in the ethmoid sinus, one mainly located in the nasal septum. According to the IRS, 1 case was at stage II, the other 4 cases were at stage III. Compared to the grey matters, on T1 weighted image, the masses were homogenously isointensity in 2 cases, isointensity with patchy hyperintensity in 3 cases. On T2 weighted images, the masses were slightly hyperintensity with patchy hypointensity in 2 cases. Slightly hyperintensity with patchy hyperintensity in 3 cases. All 5 cases were markley heterogeneously enhanced after administration of contrast agents, with patchy of non-enhanced area. Of them, two were grape-like enhanced. Four cases presented with intratumor hemorrhage. Five with bony destruction. All of the 5 cases showed orbits and anterior cranial fossa meningeal involvement.

CONCLUSIONSThere are some special MRI findings in embryonal rhabdomyosarcoma located in nasal cavity and paranasal sinuses. MRI can depict the encroachment of the tumor accurately, and may play an important role in clinical staging and in curative effect evaluation.

Adolescent ; Adult ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Nasal Cavity ; Paranasal Sinus Neoplasms ; diagnosis ; pathology ; Retrospective Studies ; Rhabdomyosarcoma, Embryonal ; diagnosis ; pathology ; Young Adult

Abdominal Pain ; etiology ; Child ; Child, Preschool ; Endoscopy, Gastrointestinal ; Female ; Humans ; Male ; Purpura, Schoenlein-Henoch ; complications ; pathology

Matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF-IMS) has been a classical technique for studying proteomics in present and a tool for analyzing the distribution of proteins and small molecules within biological tissue sections. MALDI-TOF-IMS can analyze multiple unknown compounds in biological tissue sections simultaneously through a single measurement which can obtain molecule imaging of the tissue while maintaining the integrity of cellular and molecules in tissue. In recent years, imaging mass spectrometry technique develops relatively quickly in all biomedical domain. This paper based on the relevant data and reviews the present developing level of MALDI-TOF-IMS, the principle of imaging mass spectrometry, methology and the prospect in forensic pathology.
Diagnostic Imaging ; Forensic Sciences/methods* ; Humans ; Male ; Proteins ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Abstract: Objective　To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods　The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results　RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion　The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.

Objective To evaluate the efficacy and safety of perioperative anticoagulation and antiplatelet therapy of carotid endoarterectomy (CEA). Methods A retrospective study on 110 cases (122 CEAs) of carotid stenosis between Jan 2004 and Dec 2008 was undertaken. 122 cases were divided into anticoagulation group and antiplatelet group according to the perioperative medical treatment. Postoperative results of stroke/death and wound hemotoma were compared between the two groups and statistically analyzed. Results 45 CEAs were given perioperative combination of anticoagulation and antiplatelet treatment. This comprised the anticoagulation group. The antiplatelet group consisted of the other 77 CEAs which were treated with antiplatelet solely. Perioperative stroke/death rates were equivalent (2.2% anticoagulation vs. 2.6% antiplatelet, P =0.897). Wound hemotoma rates were found with statistical significant difference between the two groups (13.3% anticoagulation group vs. 1.3% antiplatelet group, P = 0. 006 ). Conclusion Our results suggest that perioperative antiplatelet therapy in perioperative carotid endoarterectomy does not increase perioperative stroke/death risk, while perioperative anticoagulation increases the risk of wound hematoma.