Objective To explore the main reason of secondary barrenness resulted from drug miscarriage before childbearing. Methods Gynecologic examination was performed in 156 patients with secondary barrenness after drug miscarriage. Uterus neck and vaginal smear examination, mycoplasma and chlamydozoon detection, and hysterosalpingograghy were performed in the same time. Results 101 patients (101/156,64.74%) had genital duct inflammation,61 patients (61/156,39.1%) had various degrees of tubal obstruction. Conclusion The main reason of secondary barrenness after drug miscarriage was the tubal obstruction resulted from inflammation, especially chlamydozoon and mycoplasma infection. Drug miscarriage was not so safe before childbearing.

Objective To explore a new method for one-stage repair of the intestinal leakage based on the principle of magnetic compression anastomosis. Methods Twenty-four dogs were randomly divided into experimental group (n = 12) and control group (n = 12) according to random number table. The model of upper and multiple intestinal leakages was established by making transverse incisions of 1 cm in length on the jejunum wall about 50 cm and 100 cm away from the Treitz ligament. Forty-eight hours later, two NdFeB magnetic rings with the magnetic flux of 2500 G were put into the intestine from the leak sites. The leak sites were pressed between the two rings. The ventages in the control group were sutured. The condition of the dogs was observed after the repair of the leakage. The excreting time was recorded, and the leakage pressures of the anastomotic stoma were detected.The positions of the magnetic rings in the experimental group were detected by X ray. Tissues of the anastomotic stoma were processed by hematoxylin eosin and Masson staining. All data were analyzed using the two-sample t test. Results Severe abdominal infection occurred 48 hours after the establishment of the model. All the intestinal leakages in the experimental group were successfully repaired and the dogs survived for a long time. The magnetic rings were excreted six or seven days after the repair. Eight dogs of the control group survived. The leakage pressure of the anastomotic stoma seven days after the repair was (134 ±23)mm Hg (1 mm Hg =0. 133 kPa) in the experimental group and (91 ± 18)mm Hg in the control group, respectively, with a significant difference between the two groups (t = 3.225, P ＜ 0.05). The leakage pressure of the anastomotic stoma 14 days after the repair was (281 ±7)mm Hg in the experimental group and (271 ±21) mm Hg in the control group, respectively, with no significant difference between the two groups (t =0. 988, P ＞ 0.05). Histological observation showed that after the magnetic compression anastomosis, the intestinal muscle and mucosa recovered well, inflammatory reaction was slight and less collagen fiber and scar was formed. Conclusions Application of magnetic ring with the magnetic flux of 2500 G in one-stage repair of the intestinal leakage in the state of severe abdominal infection is safe and reliable.

Objective To investigate the predictive value of plasma copeptin level for the outcomes in patients with acute ischemic stroke. Methods Consecutive patients with acute ischemic stroke were enroled in the study. Enzyme-linked immunosorbent assay was used to detect the plasma copeptin level. The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate baseline stroke severity. The outcome was evaluated at 90 days with the modified Rankin Scale (mRS), and the good outcome was defined as mRS 0 - 2. Results A total of 160 patients with acute ischemic stroke were enroled, 121 had good outcome and 39 had poor outcome. The age (71. 87 ± 6. 11 years vs. 66. 19 ± 9. 39 years; t =- 3. 540, P = 0. 001), serum levels of C-reactive protein (6. 84 ± 2. 80 mmol/L vs. 5. 84 ± 2. 89 mmol/L;t = - 2. 459, P = 0. 023) and copeptin (143. 12 ± 34. 02 pmol/L vs. 50. 78 ± 18. 62 pmol/L; t = 21. 564, P <0. 001), NIHSS score (12. 00 ± 4. 00 vs. 6. 00 ± 3. 00; t = - 7. 861, P < 0. 001), as wel as proportions of patients with hypertension (79. 5% vs. 60. 3% ; χ2 = 4. 758, P = 0. 029), atrial fibrilation (20. 51% vs. 7. 44% ; χ2 = 4. 022, P = 0. 045), and large artery atherosclerotic stroke (43. 59% vs. 22. 31% ; χ2 = 6. 696, P = 0. 010) in the poor outcome group were significantly higher than those in the good outcome group, but diastolic blood pressure was significantly lower (89 ± 12 mmHg vs. 95 ± 9 mmHg, 1 mmHg = 0. 133 kPa;t = 3. 323, P = 0. 001). Multivariate logistic regression analysis showed that the plasma copeptin level (odds ratio 2. 332, 95% confidence interval 1. 725 - 3. 153; P < 0. 001) was an independent risk factor for the poor outcome in patients with acute ischemic stroke. Person correlation analysis showed that the plasma copeptin level and baseline NIHSS score showed a significant positive correlation (r = 0. 895, P < 0. 001). Receiver operating characteristic (ROC) analysis showed that plasma copeptin level has a significant predictive value for the poor outcome at day 90 after acute ischemic stroke (area under the ROC curve = 0. 740, 95%confidence interval 0. 623 - 0. 783; P < 0. 01). When plasma copeptin level > 104. 3 pmol/L was used as the cutoff value, the sensitivity and specificity for predicting the poor outcomes at day 90 after onset were 86. 8% and 40. 2% , respectively. Conclusions The plasma copeptin level may be a good predictor for neurological outcome at day 90 after onset in patients with acute ischemic stroke.

AIM: To investigate the single nucleotide polymorphisms (SNPs) in the METTL4 gene which was mapped to 18p11.31, and the relationship between the SNPs and high myopia. METHODS: Genomic DNA was collected from 71 control subjects and 177 individuals with high myopia. Among them, there were 59 autosomal dominant high myopia probands (AD group), 46 autosomal recessive probands (AR group) and 72 patients non-transmitted (SF group). The exons of METTL4 gene were analyzed by polymerase chain reaction, heteroduplex-single strand conformation polymorphism (HA-SSCP) and sequencing. RESULTS: There were 2 SNPs of METTL4 gene in high myopia individuals and control subjects: SNP7438A→C, Glu230Asp, which hadn't been reported in GenBank;and SNP131C→A, Gln310Lys. SNP7438A→C genotypes between controls and high myopia groups were not different. SNP131C→A genotypes between controls and AR or SF groups were not different, while SNP131C→A genotypes showed a significant difference between AD group and control subjects. CONCLUSION: In METTL4 gene, SNP7438A→C is not responsible for high myopia. Further studies are needed to confirm whether SNP131C→A is responsible for autosomal dominant high myopia.

BACKGROUND: Experimental evidence suggests that growth factors can promote myocardial angiogenesis, but the effect and mechanism of basic fibroblast growth factor (bFGF) in controlled release delivered via fibrin glue has not been fully recognized.OBJECTIVE: To evaluate the effect of controlled-release bFGF delivered via fibrin glue in the myocardium on the expressions of vascular growth factors and cell proliferation in the local acute myocardial infarct area in canines, and assess the therapeutic effect of this strategy.DESIGN: Completely randomized controlled experiment.SETTING: Department of Cardiology, Beijing Anzhen Hospital, Capital University of Medical Sciences; Department of Cardiology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology; Shanghai Xinxing Blood Product Research Institute.MATERIALS: This experiment was carried out in the Laboratory of Animal Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, and the Experimental Animal Center of Chinese PLA General Hospital between June 2001 And March 2003.Twelve clean healthy adult mongrel dogs of either sex were selected and randomized into transmyocardial laser revascularization group and bFGF group with 6 in each group.METHODS: With appropriate anesthesia, the chest of the dog was opened and the left anterior descending (LAD) branch of the coronary artery was ligated to establish acute myocardial infarction (AMI) model.The dogs were then randomized into transmyocardial laser revascularization group to receive transmural myocardial penetration 30 minutes after AMI and bFGF group with non-transmural myocardial penetration 30 minutes after AMI and subsequent injection of bFGF-containing fibrin glue into the channel. The expressions of vascular epithelial growth factor (VEGF), transforming growth factor beta 1 (TGFβ1) and proliferating cell nuclear antigen (PCNA) in the loacl ischemic myocardium were examined immunohistochemically (IHC) at postoperative 18 weeks.MAIN OUTCOME MEASURES: Quantitative IHC analysis of VEGF,TGFβ1 and the PCNA expressions in the local ischemic myocardia in transmyocardial laser revascularization group and bFGF group.RESULTS: Five dogs in the transmyocardial laser revascularization group and 6 in the bFGF group survived the operations. Quantitative IHC analysis revealed obviously larger positive area stained for myocardial VEGF,TGFβ1 and PCNA in bFGF group than in transmyocardial laser revascularization group (t=-7.505, -2.690 and -6.895, P ＜ 0.05), and the average absorbance of PCNA staining in bFGF group was greater than that in the transmyocardial laser revascularization group (t= -5.271, P ＜ 0.05).CONCLUSION: Controlled-releasing bFGF delivered in the myocardium can increase local expressions of the vascular growth factors in the ischemic myocardium and enhance cell proliferation, promoting revascularization after AMI.

The extracelluar domain I-IV of target gene VEGFR2 (Vascular endothelial growth factor receptor 2) was cloned from villus of trimester abortion by RT-PCR, and linked to the expression vectors. Then, the transfection conditions were optimized in serum-free suspension culture HEK293 using GFP (Green fluorescence protein) as the report gene. The results showed that the optimal transfection efficiency and cell number were obtained when the ratio of foreign DNA: PEI = 1:2 (W/W), DNA = 1.5 g /10(6) cells and shaking speed (120 r/min) in serum free medium in the beginning 4 hours of transfection. After optimizing the transfection conditions, the expression vector was successfully constructed for transient gene expression in HEK293, COS-7, and CHO-K1. The result shows that the target protein was only detected in CHO-K1 supernatant. Because of the C-terminal 8-His tag of target protein, target protein was subsequently purified using Ni2+-IDA and 5 mg purified protein was obtained in 1.5 L supernatant of CHOK1.
Animals ; CHO Cells ; Cell Line ; Chorionic Villi ; metabolism ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Culture Media, Serum-Free ; Gene Expression Regulation ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Solubility ; Transfection ; Vascular Endothelial Growth Factor Receptor-2 ; biosynthesis ; genetics

Objective:This study aims to discuss the diagnosis and treatment of pediatric appendicovesical fistula (AVF).Methods:A retrospective analysis was conducted on the clinical data of a pediatric patient with AVF admitted to our hospital in March 2023. The patient was a 6-year and 11-month old male who was hospitalized on March 21, 2023, due to difficulty urinating accompanied by diarrhea for two weeks. Computed tomography (CT) revealed bladder stones. The preoperative diagnosis was bladder stones. Transurethral cystoscopic lithotripsy with laser was performed under general anesthesia. Two weeks postoperatively, the child presented with recurrent symptoms of frequent urination, urinary pain, and diarrhea. Urine routine examination indicated a urinary tract infection. Over a month of antibiotic treatment was ineffective, and symptoms such as pneumaturia and fecaluria emerged, with exacerbation of diarrhea, suggesting the possibility of a fistulous tract between the child's intestine and bladder. Further bladder ultrasonography with contrast showed microbubbles of contrast medium leaking from the right posterior bladder wall into the intestinal tract. Enhanced magnetic resonance imaging (MRI) demonstrated a small, sharp tube-like shadow at the upper edge of the right posterior bladder, with a strip-like, significantly enhanced shadow within the lumen. The preoperative diagnosis was revised to appendicovesical fistula. During cystoscopic examination, a papillary-like protrusion was identified on the right lateral wall of the bladder, with no evident orificium fistulae or foreign body discharge noted at the protrusion site. Consequently, robot-assisted laparoscopic partial cystectomy, appendectomy, and lysis of adhesions were performed.Results:The patient was administered antibiotic for a 10-day course of anti-infection and a urinary catheter was maintained for 13 days. The patient recovered entirely and had been discharged after the removal of the urinary catheter. At an 11-month follow-up, there were no reported specific discomforts.Conclusions:Pediatric AVF is rare, and bladder contrast-enhanced ultrasonography and MRI are preferred for initial diagnostic evaluation. The diagnosis can be confirmed by specific clinical presentations such as intermittent pneumaturia and fecaluria, diarrhea with bladder stones. Laparoscopic surgery or robot-assisted laparoscopic surgery could be a feasible treatment option.

Objective:To establish a colloidal gold technique assay for the rapid detection of immunoglobulin(Ig)M and IgG antibodies against 2019 novel coronavirus (2019-nCoV) and to evaluate its clinical performance.Methods:A total of 278 patients who were respectively treated at Wuhan Hankou Hospital and the People′s Hospital of Honghu from February 12, 2020 to February 20, 2020 were collected. According to the diagnostic criteria, 89 patients were confirmed with positive 2019-nCoV nucleic acid, and 189 were 2019-nCoV nucleic acid-negative suspected patients. A total of 273 medical examiners from Nanfang Hospital, Southern Medical University from 2015 to 2018 were selected as controls. The serum samples of patients were collected. 2019-nCoV nucleic proteins were obtained from prokaryotic expression vectors. Indirect IgM and IgG colloidal gold techniques were established by using recombinant nuclear protein. 2019-nCoV nucleic acid detection by reverse transcription-polymerase chain reaction (RT-PCR) was used as control. Serum specimens were tested for 2019-nCoV IgM and IgG. The specificity and sensitivity of colloidal gold assay were analyzed.Results:The positive rates of IgM and IgG with the colloidal gold detection in confirmed patients with positive 2019-nCoV nucleic acid were 78.7%(70/89) and 73.0%(65/89), respectively. The positive rates of IgM and IgG in medical examiners were 1.8%(5/273) and 0.7%(2/273), respectively. The sensitivity and specificity of IgM detection reagents were 78.7% and 98.2%, respectively, those of IgG detection reagents were 73.0% and 99.3%, respectively, and those of IgM combined with IgG detection were 87.6% and 98.2%, respectively. For suspected patients with negative 2019-nCoV nucleic acid, the positive rates of IgM and IgG were 59.8%(113/189) and 52.9%(100/189), respectively, and the positive rate of IgM combined with IgG detection was 66.1%(125/189).Conclusion:This reagent of 2019-nCoV antibodies detection (colloidal gold technique) fulfills the requirement for clinical application with high specificity and sensitivity, which can be served as a supplementary detection method for 2019-nCoV nucleic acid detection by RT-PCR.

Objective: To establish a colloidal gold technique assay for the rapid detection of immunoglobulin(Ig) M and IgG antibodies against 2019 novel coronavirus (2019-nCoV) and to evaluate its clinical performance. Methods: A total of 278 patients who were treated at Wuhan Hankou Hospital and the People's Hospital of Honghu from February 12, 2020 to February 20, 2020 were collected. According to the diagnostic criteria, 89 patients were confirmed with 2019-nCoV nucleic acid positive diagnosis, and 189 were 2019-nCoV nucleic acid-negative suspected patients. A total of 273 medical examiners from Nanfang Hospital, Southern Medical University from 2015 to 2018 were selected as controls. The serum samples of patients were collected. 2019-nCoV nucleic proteins were obtained from prokaryotic expression vectors. Indirect IgM and IgG colloidal gold techniques were established by using recombinant N protein. 2019-nCoV nucleic acid detection by reverse transcription-polymerase chain reaction (RT-PCR) was used as control. Serum specimens were tested for 2019-nCoV IgM and IgG. The specificity and sensitivity of colloidal gold assay were analyzed. Results: The sensitivity and specificity of IgM detection reagents were 78.7% and 98.2%, respectively, those of IgG detection reagents were 73.0% and 99.3%, respectively, and those of IgM combined with IgG detection were 87.6% and 98.2%, respectively. For suspected patients with negative 2019-nCoV nucleic acid, the positive rates of IgM and IgG were 59.8% (113/189) and 52.9% (100/189), respectively, and the positive rate of IgM combined with IgG detection was 66.1% (125/189). Conclusion This reagent of 2019-nCoV antibodies detection (colloidal gold technique) fulfills the requirement for clinical application with high specificity and sensitivity, which can be served as a supplementary detection method for 2019-nCoV nucleic acid detection by RT-PCR.