AIM To investigate if scallop (Placopecta magellanicus) skirt glycosaminoglycan (SS-GAG) inhibits the proliferation of vascular smooth muscle cell (VSMC) as heparin does so and to clarify its mechanism. METHODS The inhibitory effects of SS-GAG on the proliferation of rat thoracic aorta and abdominal aorta VSMC induced by fetal bovine serum (FBS) or basic fibroblast growth factor (bFGF) were determined by cell counting, crystal violet staining and MTT colorimetry. The effects of SS-GAG on the expression of proliferating cell nuclear antigen (PCNA) and platelet-derived growth factor (PDGF) in VSMC proliferation induced by bFGF were evaluated by immunohistochemical technique (LSAB method) and computer image analysis system. RESULTSSS-GAG exerted antagonistic effects on VSMC proliferation induced by 20% FBS and 50 μg·L-1 bFGF at concentrations ranging from 50 mg·L-1 to 200 mg·L-1 and repressed the increasing expression of PCNA and PDGF. CONCLUSION SS-GAG significantly inhibits the proliferation of VSMC, which may be carried out through repression of PDGF and PCNA expression.

BACKGROUND: Hepatocyte growth factor (HGF) promotes neurite outgrowth from neocortical explants, and supports neuronal survival under serum-free condition. Thus, HGF can mediate neurotrophic function as a novel neurotrophic factor.OBJECTIVE: To establish an in vitro injury model with a semi-solid culture system for the purpose of improving the evaluation of neurite regeneration of transected spinal cord neurons from rat embryo, and investigate the effect of HGF on neurite regeneration.DESIGN: Randomized controlled study.SETTING: Hematology Laboratory of Radian Medical Institute of Academy of Military Medical Sciences of PLA.MATERIALS: The experiment was carried out at the Hematology Laboratory of Radian Medical Institute of Academy of Military Medical Sciences of PLA from August 2004 to May 2005. Wistar fetal rats of 14-16 days old were provided by Animal Center of Academy of Military Medical Sciences of PLA. Tail collagen was extracted from adult male Wistar rats with body mass of (250±50) g.METHODS: ① Rat tail type Ⅰ collagen substrate was prepared and spread on a culture dish, cut into about 0.5-1.0 mm3 slices, then spinal cord slices of 15-day-old fetal Wistar rat were explanted on the primary culture. Five days later, the outgrowing processes were severed, then a block of collagen, with the surface area of 2 mm2 and 200 μm away from the slice, was removed and the vacancy was replaced with a fresh collagen block of 2 μL after aspirating the medium. The fresh collagen block could be solidified and then fresh liquid medium was added as the secondary culture. The regeneration of neurite was observed by microscopy at 0, 1, 6,12 and 24 hours after severing. ② The medium was changed with 0.5% N3-conditioned medium. 10 μg/L HGF was added in the experimental group, and 0.5% N3-conditioned medium was added in the control group.The status of regeneration was evaluated by the average value of 3 longest regenerative neurites for each slice. There were 12 slices in each group.The status of neurite regeneration was calculated and was evaluated 24 hours later.MAIN OUTCOME MEASURES: ① neurite regeneration in situ; ②comparisons of neurite regeneration between control group and experimental group.RESULTS: ① Neurite regeneration in situ: The neurites disintegrated near the severing line immediately following the transection injury. This process persisted about 1-2 hours and the distance away from the severing line was about 20 μm. Then the proximal end of neurites would swell and thicken. At this time neurites stopped collapsing and neurite regeneration began. Their regenerating rate would quicken at 12 hours after severing. Regenerating neurites were more branching and curlier as compared with original neurites. ② Comparisons of neurite regeneration between control group and experimental group: The average length of regenerative neurites was more in the experimental group than that in the control group [(375±96) μm, (200±75) μm, P ＜ 0.05].CONCLUSION: ① We establish a simple, economic model to evaluate neurite regeneration. ② By this model, we prove that HGF can promote neurite regeneration.

Objective:To observe the correlation between the B7 blocker,CD28 and cytotoxic T lymphocyte-associated antigen 4 immunoglobulin (CTLA-4Ig) impacting on Th17 and Treg cells expressed in the spleen of lupus mice and its intervention role in lupus-like symptoms of B6.MRL-Faslpr/J lupus mice.Methods:Sixteen 4-month-old female B6.MRL-Faslpr/J mice were selected and randomly divided into treatment group ( groupⅠ) and control group ( groupⅡ);injected the same amount of CTLA-4Ig and PBS intra-venously,checked their 24 hour urine protein ,ANA antibody,ds-DNA antibodies before and after the intervention.Two weeks after the intervention ,detected serum IL-17A,and the percentage of Th17 and Treg cells in their spleen.Results:Two weeks after the last inter-vention,24-hour urine protein,serum ANA and ds-DNA in groupⅠdecreased,and all the differences were statistically significant (P<0.05) compared with groupⅡ.Two weeks after the last intervention,serum IL-17A and the proportion of Th17 cells in the spleen in groupⅠwere lower than those in groupⅡ, but Treg cells in CD4+T lymphocytes was higher than that in groupⅡ,all the differences were statistically significant ( P<0.05).Conclusion:CTLA4-Ig can relieve the lupus-like symptoms in lupus mice;raising the number of Treg cells and decreasing the number of Th17 cells may be one of the important mechanisms for CTLA-4Ig to alleviate lupus-like symptoms in B6.MRL-Faslpr/J mice.

Objective To investigate the mRNA expression of toll like receptor-9 (TLR9) and interferon regulatory factors-5 (IRF5) of AS2O3 on peripheral blood mononuclear cells (PBMCs) in systemic lupus erythematosus (SLE) patients.Methods PBMCs of 15 SLE patients and 15 healthy subjects were treated with different concentrations of AS2O3 and cyclophosphamide (CTX) in vitro.Real-time quantitative polymerase chain reaction was used to amplify TLR9 and IRF5 gene before and after 12 and 24 hours drug intervention and the mRNA expressions were measured.Differences between groups were analyzed by paired t test or variance analysis.Results The mRNA expression levels of TLR9 [12 h(1.38±0.26) and 24 h (1.28±0.35)] on PBMCs in SLE patients were significantly higher than those in healthy controls [12 h(1.05±0.35) and 24 h (0.97±0.19)](t=2.37,P=0.03; t=2.44,P=0.02).The IRF5 mRNA expression levels [12 h (0.95±0.27) and 24 h (0.91 ±0.35)] in SLE patients were obviously higher than those in healthy controls [12 h (0.62 ±0.23) and 24 h (0.60±0.39)] (t =3.07,P=0.01 ; t =3.45,P＜0.01).AS2O3 could suppress the mRNA expression of TLR9 on PBMCs and the effect was gradually increasing with the increasing concentration of AS2O3 and processing time [0.2 mg/L AS2O3 group 12 h (0.430±0.110) and 24 h(0.290±0.050),0.4 mg/L AS2O3 group 12 h (0.170±0.038) and 24 h (0.090±0.017),0.8 mg/L AS2O3 group 12 h (0.023±0.011) and 24 h (0.003±0.001)].Comparing with CTX [12 h (0.814±0.081) and 24 h(0.755±0.139)],AS2O3 had a more significant strong effect on inhibiting the expression of TLR9 mRNA in SLE patients [F=165.32(12 h),P＜0.01; F=99.20 (24 h),P＜0.01].The mRNA expression of IRF5 on PBMCs was not suppressed by AS2O3 and CTX and there was no statistically significant difference between groups (P＞0.05).Conclusion There is abnormal expression of IRF5 and TLR9 mRNA in SLE patients.AS2O3 may suppress the TLR9 mRNA expression in SLE patients,which may be one mechanism of clinical effectiveness.

Objective:To evaluate the hemostatic effect of self-expanding polyurethane foam in an animal model of fatal hepatic trauma and hemorrhage.Methods:The fatal liver trauma hemorrhage model with swine was established. Then the damage-controlled resuscitation was performed. Thirty minutes after injury, the experimental animals were randomly divided into the gauze packing group (GP), foam packing group (FP) and blank control group (BC). The survival time, vital signs, the bleeding volume, coagulation function and other lab indicators were recorded for 48 h. Liver histopathological examination was performed after death or execution.Results:All the three groups had severe hemorrhagic shock after modeling. The 48-h survival rate of the FP group and the GP group was significantly higher than that of the BC group (6/6 vs 4/6 vs 0/6). The average survival time of the FP group was not statistically different from that of the GP group [48 h vs (44.58±5.53) h, P>0.05], and was significantly longer than that of the BC group [48 h vs (1.64±0.17) h, P<0.01]. The bleeding volume of the FP group was significantly less than the GP group and BC group [(19.2±7.3) g/kg vs (41.3±8.6) g/kg, (51.5±7.3) g/kg, both P<0.01]. Compared with the GP group and the BC group, the cardiac output of the FP group was significantly improved [(5.00±0.53) L/min vs (4.13±0.41) L/min, (2.38±0.48) L/min, both P<0.05]. The coagulation function, liver and kidney function and blood lactate level of the FP group and the GP group were better than those of the BC group; the intra-abdominal pressure of the FP group was significantly higher than that in the GP group [(18.83±3.25) cmH 2O vs (3.83±1.47) cmH 2O, P<0.05]. There was no abnormal increased in intra-abdominal pressure in the BC group. According to the histopathology examination, there was no obvious secondary damage in the FP group. Conclusions:The application of self-expanding polyurethane foam for intraperitoneal packing to stop bleeding can effectively reduce the amount of bleeding in the fatal liver trauma hemorrhage model, effectively maintain vital signs, and improve the short-term survival rate.

Objective:To investigate the influencing factors associated with lesion residual after minimally invasive rotary mastectomy for breast masses and to construct a predictive model using columnar plots.Methods:Two hundred and twenty-eight patients with Breast lumps in Linyi People’s Hospital from Jun. 2018 to Jun. 2020 were selected as study subjects, all of whom underwent minimally invasive rotational resection, and the influencing factors of postoperative lesion residual were analyzed using univariate and multifactorial analysis, and a column line graph risk warning model was constructed and given for evaluation and validation.Results:228 patients were followed up for 6 months after surgery, 3 cases were lost, 225 cases completed postoperative follow-up, among which 185 cases (82.22%) had successfully resected lesions; 40 cases (17.78%) had residual lesions. Univariate, logistic regression analysis showed that tumor diameter ( t=15.52, P<0.001) , lesion morphology ( t=15.52, P<0.001) , lesion boundary ( χ2=7.71, P=0.006) , ultrasound guidance ( χ2=12.69, P<0.001) , and minimally invasive spinotomy system ( χ2=10.64, P=0.001) were the factors influencing lesion residual after minimally invasive spinotomy of breast masses. Based on the above factors to construct a column line graph model of lesion residual after minimally invasive rotational mastectomy for breast lumps, the area under the working characteristic curve (ROC) for model subjects was 0.974 (95% CI: 0.954-0.994) , indicating that the calibration curve basically matched the actual curve; external validation of the model showed that the area under the obtained ROC was 0.962 (95% CI: 0.933-0.991) , indicating that the model has a high degree of calibration. Conclusion:Tumor diameter, lesion morphology, lesion boundary, ultrasound guidance, and minimally invasive spinotomy system are the influencing factors of lesion residual after minimally invasive spinotomy for Breast lumps, and the column line graph model prediction of lesion residual occurring after surgery constructed based on the above factors has good confidence.

OBJECTIVETo explore the clinical and gene mutation characteristics of a child with maturity-onset diabetes of the young 2 (MODY2).

METHODThe clinical and follow-up data of 1 patient with MODY2 were reviewed. GCK mutational analysis was performed by PCR and direct sequencing in the proband and his family members.

RESULTThe 9 years and 6 months old boy was referred to our department for short stature and mild hyperglycemia. His fasting blood glucose was elevated to 7.4-7.8 mmol/L, hemoglobin A1C 6.7%. His height was 122 cm (-2 s), weight 25 kg (-1 s), body mass index (BMI) 16.8 kg/m(2). His physical exam was unremarkable without dysmorphic features or acanthosis nigricans. The oral glucose tolerance test (OGTT) showed fasting glucose 8.17 mmol/L, insulin <2.0 mU/L, 2 h glucose 8.69 mmol/L, insulin 5.06 mU /L. The boy was treated with insulin injection for half a year. His fasting blood glucose was stable at 5.6-8.5 mmol/L, hemoglobin A1C 6.7%-6.8%. His mother's fasting blood glucose was 6.86 mmol/L, OGTT 2 h blood glucose 10.36 mmol/L, hemoglobin A1C 6.8%. GCK sequence revealed a novel GCK mutation c.34_44+15del26 in the proband and his mother, which was co-segregated with diabetes. The boy's treatment was shifted from insulin injection to diet and exercise after the diagnosis of MODY2 was confirmed. Being followed up for 2 and a half years, his fasting blood glucose was stable at 4.6-8.0 mmol/L and hemoglobin A1C 6.8%-7.1%.

CONCLUSIONThe clinical features of MODY2 are persistent and stable fasting hyperglycemia over a period of months or years and small blood glucose increment (less than 3 mmol/L) after an OGTT (2 h glucose-fasting glucose). We identified a novel c.34_44+15del26 mutation in GCK which co-segregated with diabetes phenotype in this family.

Asian Continental Ancestry Group ; genetics ; Blood Glucose ; Child ; Diabetes Mellitus, Type 2 ; diagnosis ; genetics ; Fasting ; Follow-Up Studies ; Glucokinase ; genetics ; Glucose Tolerance Test ; Glycated Hemoglobin A ; Humans ; Hyperglycemia ; Insulin ; Male ; Mutation ; Phenotype

To investigate the clinical manifestations, molecular genetics and gonadal pathology characteristics of patients with disorders of sex development (DSD), and to summarize the clinical experience of identifying rare diseases from common symptoms.

The clinical data of 416 patients with DSD diagnosed and treated in the multidisciplinary center of DSD of Guangzhou Women and Children′s Medical Center from May 2018 to August 2023 were retrospectively analyzed, summarized and discussed.

According to chromosome karyotype, 416 cases of DSD were classified into three types: 92 cases(22.1%) of abnormal sex chromosome karyotype, 285 cases(68.5%) of 46, XY karyotype and 39 cases(9.4%) of 46, XX karyotype. Among the 92 patients with abnormal sex chromosome karyotype, 59 cases were raised as males, 18 cases (30.5%) complained of short penis with hypospadias and cryptorchidism. The most common karyotype was 45, X/46, XY(58 cases, 63.0%).Among the 285 patients with 46, XY karyotype, 238 cases were raised as males, and 63 cases(26.5%)complained of short penis and hypospadias; 47 cases were raised as females, and 13 cases(27.7%) complained of inguinal mass. A total of 216 patients with 46, XY karyotype were subjected to whole exome gene detection, and 155 cases(71.8%) were found to have molecular pathogenesis with the clinical phenotype. Among the 39 patients with 46, XX karyotype, 19 cases were raised as males, and 8 cases(42.1%) complained of short penis and hypospadias. In the 18 cases of gonad biopsy, 17 cases showed testicular tissue in gonads. Whole exome sequencing was performed in 14 cases. NR5A1 gene heterozygous mutation, SRY gene mutation and SOX3 gene mutation were found in 2 cases, respectively(14.3%). Twenty cases were raised as females, and 14 cases(70.0%) complained of clitoral hypertrophy. Gonad biopsy was performed in 8 cases, with 7 cases of ovotestis(87.5%) and 1 case of NR5A1 gene heterozygous mutation(14.3%).

The etiologies of DSD are complex and diverse, and the clinical manifestations are various, which can be manifested as hypospadias, micropenis, cryptorchidism and other common symptoms of the urinary system. Different etiologies have different treatment options. Therefore, chromosome karyotype, molecular genetic testing and gonadal pathology can be used to clarify the cause of disease, especially for rare diseases, improve the detection rate, reduce the rate of missed diagnosis, and ensure reasonable treatment, especially sex selection.