Whole-genome sequencing analysis in fetal structural anomalies: novel phenotype-genotype discoveries.

OBJECTIVES: The identification of structural variants and single-nucleotide variants is essential in finding molecular etiologies of monogenic genetic disorders. Whole-genome sequencing (WGS) is becoming more widespread in genetic disease diagnosis. However, data on its clinical utility remain limited in prenatal practice. We aimed to expand our understanding of implementing WGS in the genetic diagnosis of fetal structural anomalies. METHODS: We employed trio WGS with a minimum coverage of 40× on the MGI DNBSEQ-T7 platform in a cohort of 17 fetuses presenting with aberrations detected by ultrasound, but uninformative findings of standard chromosomal microarray analysis (CMA) and exome sequencing (ES). RESULTS: Causative genetic variants were identified in two families, with an increased diagnostic yield of 11.8% (2/17). Both were exon-level copy-number variants of small size (3.03 kb and 5.16 kb) and beyond the detection thresholds of CMA and ES. Moreover, to the best of our knowledge, we have described the first prenatal instance of the association of FGF8 with holoprosencephaly and facial deformities. CONCLUSIONS: Our analysis demonstrates the clinical value of WGS in the diagnosis of the underlying etiology of fetuses with structural abnormalities, where routine genetic tests have failed to diagnose. Additionally, the novel variants and new fetal manifestations have expanded the mutational and phenotypic spectrums of BBS9 and FGF8. © 2023 International Society of Ultrasound in Obstetrics and Gynecology.

[Clinical value of ultrasound-guided radiofrequency ablation in the treatment of retroperitoneal tumors].

Objective: To explore the clinical value of ultrasound-guided radiofrequency ablation in the treatment of retroperitoneal tumors. Methods: The clinical data of 13 patients with retroperitoneal tumors treated with ultrasound-guided radiofrequency ablation in the First Affiliated Hospital of Zhengzhou University from January 2018 to January 2020 were analyzed retrospectively. The ablation effect was evaluated and the postoperative complications were observed. The changes of tumor volume before and after radiofrequency ablation were compared. Results: The symptoms of pain and dyspepsia were significantly improved after radiofrequency ablation, and the hospital stay was (9.2±2.9) days. The tumor was ablated completely in 10 cases, tumor residual in 1 case and tumor metastasis in 2 cases. One patient had postoperative duodenal perforation complicated with intra-abdominal infection, and no serious complications occurred in other patients. There were 20 lesions in 13 patients. The maximum diameter of 20 lesions before operation and 1, 3, 6 months after operation were (39.5±15.9) mm, (30.6±4.9)mm, (15.6±7.7) mm and (9.9±3.1) mm, respectively, the maximum diameters of 1, 3 and 6 months after operation were smaller than that before operation (P<0.05). Conclusion: Ultrasound-guided radiofrequency ablation is a real-time, accurate, safe and effective minimally invasive treatment with few complications, and has a high clinical value for retroperitoneal tumors.

[Efficacy and initial clinical evaluation of optical genome mapping in the diagnosis of structural variations].

To investigate the efficacy and value of optical genome mapping (OGM) in detecting chromosomal structural variations. In a clinical study about high-precision analysis of genomic structural variation for complex genetic diseases, a retrospective study was performed on the cases with karyotyping at the department of Obstetrics and Gynecology, and Endocrinology of Peking Union Medical College Hospital from January to December 2021. Ten cases with abnormal karyotype was detected by OGM. Partial cases were verified by fluorescence in situ hybridization (FISH), SNP array or CNV-seq. Results of ten cases, nine were detected with abnormality by OGM, including unbalanced chromosomal rearrangements (n=3), translocation (n=5) and paracentric inversion (n=1), and the results were in concordance with other standard assays. However, one case with breakpoint and reconnected at centromere has not been detected. In conclusion, ten samples were comprehensively analyzed by karyotyping, FISH, SNP array or CNV-seq, and OGM, and results demonstrated that optical genome mapping as a new technology can not only detect unbalanced rearrangements such as copy number variants as well as balanced translocations and inversions, but more importantly, it can refine breakpoints and orientation of duplicated segments or insertions. So it can contribute to the diagnosis of genetic diseases and prevent birth defect. However, the current technology is not yet capable of detecting breakpoints of balanced structural variations lying within unmapped regions.

Skin Ultrastructure and the Changes of HIF-2α, H-FABP Expression in the Myocardium of Electric Shock Death Rats.

ABSTRACT: Objective To observe the skin ultrastructure change of electric shock death rats and to test the expression changes of hypoxia-inducible factor-2α （HIF-2α） and heart type-fatty acid-binding protein （H-FABP） of myocardial cells, in order to provide basis for forensic identification of electric shock death. Methods The electric shock model of rats was established. The 72 rats were randomly divided into control group, electric shock death group and postmortem electric shock group. Each group was divided into three subgroups, immediate （0 min）, 30 min and 60 min after death. The skin changes of rats were observed by HE staining, the changes of skin ultrastructure were observed by scanning electron microscopy, and the expression of HIF-2α and H-FABP in rats myocardium was tested by immunohistochemical staining. Results The skin in the electric shock death group and postmortem electric shock group had no significant difference through the naked eye or by HE staining. Under the scanning electron microscope, a large number of cellular debris, cells with unclear boundaries, withered cracks, circular or elliptical holes scattered on the cell surface and irregular edges were observed. A large number of spherical foreign body particles were observed. Compared with the control group, the expression of HIF-2α in all electric shock death subgroups increased, reaching the peak immediately after death. In the postmortem electric shock group, HIF-2α expression only increased immediately after death, but was lower than that of electric shock death group （P<0.05）. Compared with the control group, the expression of H-FABP in all subgroups of electric shock death group and postmortem electric shock group significantly decreased. The expression of H-FABP in all subgroups of electric shock death group was lower than that of the postmortem electric shock group （P<0.05）. Conclusion Electric shock can increase HIF-2α expression and decrease H-FABP expression in the myocardium, which may be of forensic significance for the determination of electric shock death and identification of antemortem and postmortem electric shock.

[Risk factors for multiple debridements of the patients with deep incisional surgical site infection after spinal surgery].

OBJECTIVE: To investigate the risk factors that contribute to multiple debridements in patients suffering from deep incisional surgical site infection after spinal surgery and advise medical personnel to pay special attention to these risk factors. METHODS: We retrospectively enrolled 84 patients who got deep incisional surgical site infection after spinal surgery from Jan. 2012 to Dec. 2017. The infections occurred within 30 days after the surgery, and the identification met the criteria of deep incisional surgical site infection of Centers of Disease Control (CDC). Early debridement with first stage closure of the wound and a continuous inflow-outflow irrigation system was used, and reasonable antibiotics were chosen according to the bacterial culture results. During the treatment, the vital signs, clinical manifestations, blood test results, drainage fluid colour and bacterial culture results were acquired. If the infection failed to be controlled or relapsed, a second debridement was performed. Of the 84 cases, 60 undergwent single debridement which included 36 male cases and 24 female cases, and the age ranged from 36 to 77 years, with a mean of 57.2 years. Twenty four had multiple debridements (twice in 14 cases, three times in 6 cases, four times in 1 case, five times in 2 cases, six times in 1 cases) which included 17 male cases and 7 female cases, and the age ranged from 21 to 70 years, with a mean of 49.5 years. Risk factors that predispose patients to multiple debridements were identified using univariate analysis. Risk factors with P values less than 0.05 in univariate analysis were included together in a multivariate Logistic regression model using back-forward method. RESULTS: Multiple debridements were performed in 28.6% of all cases. The hospital stay of multiple debridements group was (82.4±46.3) days compared with (40.4±31.5) days in single debridement group (P=0.018). Instrumentation was removed in 6 cases in multiple debridements group and 4 cases in single debridement group (P=0.049). Flap transplantation was performed in 7 cased in multiple debridements group while none in single debridement group (P < 0.001). Diabetes, primary operation duration longer than 3 hours, primary operation blood loss more than 400 mL, bacteriology examination results, distant site infection were significantly different between the two groups in univariate analysis. In multivariate analysis, primary operation duration longer than 3 hours (OR=3.60, 95%CI: 1.12-11.62), diabetes (OR=3.74, 95%CI: 1.06-13.22), methicillin-resistant Staphylococcus aureus (MRSA) infected (OR=16.87, 95%CI: 2.59-109.73) were the most important risk factors related to multiple debridements in the patients with deep incisional surgical site infection after spinal surgery. CONCLUSION: Diabetes, primary operation duration more than 3 hours, MRSA infected are independent risk factors for multiple debridements in patients suffering from deep incisional surgical site infection after spinal surgery. Special caution and prophylaxis interventions are suggested for these factors.

[Impact of maternal X chromosome aneuploidies on cell free DNA prenatal screening].

Objective: To investigate the impact of maternal X chromosome aneuploidies on cell free DNA (cf-DNA) prenatal screening. Methods: After genetic counseling, invasive prenatal diagnosis was provided for the 124 cases with high risk of sex chromosome aneuploidie (SCA) indicated by cf-DNA prenatal screening. For cases with discordant results of fetal prenatal diagnosis and cf-DNA prenatal screening, maternal leukocyte was collected for copy number variation sequencing (CNV-seq) to detect whether the maternal X chromosome was carrying variations. Results: Totally, 124 cases with high risks of SCA indicated by cf-DNA prenatal screening, 9 cases refused to take invasive prenatal diagnosis, while the remaining 115 cases received. Among the 115 cases, 41 cases received accordant results with cf-DNA prenatal screening while 74 cases discordant. Among the 74 cases with discordant results, 19 cases were indicated with maternal X chromosome variations by maternal leukocyte CNV-seq, which accounting for 25.7% (19/74) of the SCA false positive cases, and 15.3% (19/124) of all SCA cases. Conclusions: Pregnant women with X chromosome variations may affect the results of cf-DNA prenatal screening, resulting in false positive or false negative outcomes, it should be emphasized that the cf-DNA results may be affected by maternal X chromosome variations. In cases with discordant results of prenatal diagnosis and cf-DNA prenatal screening, maternal leukocyte CNV-seq is recommended to find the reasons of false positive or negative results. And cf-DNA prenatal screening is not recommended for pregnant women who are already known with X chromosome variations.

[The influence of excessive daytime sleepiness and sleep quality on anxiety and depression in patients with obstructive sleep apnea hypopnea syndrome].

Objective: To explore the factors that contribute to the anxiety and depression in obstructive sleep apnea hypopnea syndrome (OSAHS) patients in terms of excessive daytime sleepiness (EDS) and sleep quality. Methods: A total of 196 OSAHS patients, including 103 severe patients and 93 mild-moderate patients, were enrolled. Polysomnography was carried on at the sleep center of the First Hospital of China Medical University between May 2013 and November 2015. According to the Epworth sleepiness scale (ESS) and the subject daytime sleepiness symptom, all patients were divided into EDS group and non-EDS group. The patients' general information and subjective symptoms were recorded. Emotional states were assessed with self-rating anxiety scale (SAS) and self-rating depression scale (SDS). Sleep quality was evaluated with Pittsburgh sleep quality index (PSQI). The anxiety and depression related factors were studied by regression analysis. Results: (1) In severe OSAHS group, the patients with EDS showed higher PSQI(6.22±2.57 vs. 4.05±3.72, P<0.01) and oxygen desaturation index(ODI)[(57.70±17.53) events/h vs. (48.23±22.01) events/h, P<0.05] when compared with those without EDS. (2) In both severe and mild-moderate OSAHS groups, the patients with EDS presented higher SAS scores (severe: 33.86±7.60 vs.28.95±4.71, mild-moderate: 37.46±10.68 vs. 33.40±11.07, P<0.05) and SDS scores (severe: 32.81±8.36 vs. 28.90±4.53, mild-moderate:36.98±12.77 vs. 31.70±10.94, P<0.05) when compared with those without EDS. (3) The multiple regression analysis showed that the SAS scores were related to ESS, PSQI, insomnia and nasal obstruction (R(2)=0.356, P<0.05), and the SDS scores were related to ESS, PSQI and insomnia(R(2)=0.344, P<0.05). Conclusions: The anxiety and depression of OSAHS patients are closely related to the severity of EDS and sleep quality. Both severe and mild-moderate OSAHS patients with EDS have worse anxiety and depression scores.

Abnormality of visual neuromagnetic activation in female migraineurs without aura between attacks.

OBJECTIVE: The present study aimed to preliminary explore the abnormal neuromagnetic activation in female migraine patients between attacks using magnetoencephalography (MEG) and pattern reversed visual evoked magnetic fields (PR-VEFs). METHODS: A total of 17 female migraine subjects during the headache-free phase and 17 healthy controls (HC) were studied using a 275-channel magnetoencephalography (MEG) system. In this study, visual evoked magnetic fields (VEFs) were generated by a pattern-reversal check as the visual stimulus. The average of 100 VEFs was evolved by different half patterns were averaged and used to analyze waveform, spectrum, and source location within two frequency ranges (5-100 and 100-1000 Hz), respectively. RESULTS: In migraine subjects, the latency of second peak of VEFs (VIIs) showed significant prolongations when compared with HC. On the sensor level, the cortical spectral power in migraine subjects was similar to that of HC in the 5-100 Hz range and was lower in the 1000-1000 Hz range. There was a decrement of source strength in the visual cortex in migraine patients when compared to HC in both the 5-100 and 100-1000 Hz frequency range. Moreover, there was a similar odds of activation in 5-100 and 100-1000 Hz frequency ranges in the area beyond the primary visual cortex between the two groups. In addition, no correlation was observed between clinical data (intensity of headache, headache-history duration, the frequency of headaches) and MEG results. CONCLUSIONS: The findings presented in the current study, suggested that interictal cortical activation following a visual stimulus was low in female migraine patients. The low pre-activation was detected in the visual cortex using VEF and MEG in both low and high-frequency band. Our results add to the existing evidence that cortical interictal excitability change may be relative to the pain-module mechanism in migraine brains. Thus, our data improved the apprehension of the cortical disorder of migraine in the high-frequency domain.

[Correlation between chest CT features and clinical characteristics of patients with bronchiectasis].

Objective: To analyze the features of chest CT imaging in adult patients with bronchiectasis and explore its correlation with clinical characteristics. Methods: From January 2010 to December 2017, patients with bronchiectasis diagnosed by chest high-resolution CT (HRCT) and aged at or above 18 years old in 5 general hospitals of Shandong province were included in the study. The correlations between the HRCT imaging features and etiology, clinical manifestations, lung function, sputum culture, prognosis and other characteristics were analyzed. Results: There were 410 bronchiectasis patients included in the study. The chest HRCT imaging of bronchiectasis were divided into three types, including columnar 46.8%, cystic 45.9% and varicose 7.3%, respectively. The HRCT imaging score was [6.0 (4.0, 7.0)]. In addition, the most common etiology of bronchiectasis was idiopathic (262, 69.3%). The proportion of idiopathic bronchiectasis in cystic bronchiectasis patients was significantly higher than that in columnar and varicose bronchiectasis (71.8% vs 58.3%, 50.0%; both P<0.017). Compared with columnar bronchiectasis, patients with cystic bronchiectasis were more likely to suffer from clinical manifestations such as cough, dyspnea, fever and wet rales (P<0.017). Compared with patients with HRCT scores of 1 to 4, patients with scores ≥8 were more likely to suffer from cough, dyspnea, fever, wet rales and clubbing (P<0.017). The proportions of pulmonary ventilatory dysfunction were significantly greater in patients with cystic bronchiectasis and varicose bronchiectasis than columnar bronchiectasis (86.7%, 86.7% vs 51.0%; both P<0.017). The HRCT scores were significantly negatively correlated with pulmonary function (P<0.001). The number of acute exacerbations, hospitalizations, and bronchiectasis severe index scores in patients with cystic bronchiectasis were significantly higher than those with columnar bronchiectasis (P<0.017). There was a significantly positive correlation between HRCT scores and the number of acute exacerbations, hospitalizations and the bronchiectasis severity index scores (P<0.001). The mortality of patients with cystic and varicose bronchiectasis was significantly higher than that of patients with columnar bronchiectasis (9.0%, 10.0% vs 2.1%; both P<0.017). Compared with patients with HRCT scores of 1 to 4, patients with scores ≥8 had a higher mortality rate (15.9% vs 0.9%; P<0.017). Conclusions: There is a correlation between HRCT findings and clinical manifestations in patients with bronchiectasis. The clinical manifestations, lung function and prognosis of patients with cystic bronchiectasis are worse than those of the columnar bronchiectasis; the higher the HRCT scores are, the worse the clinical manifestations, lung function and prognosis of the patients are.

[Clinicopathological significance of poorly differentiated clusters in colorectal adenocarcinoma].

Objective: To investigate the correlation between poorly differentiated clusters (PDCs) in colorectal adenocarcinomas with clinicopathological parameters and its clinicopathological significance. Methods: One hundred and eighty-three colorectal adenocarcinomas resected by radical proctocolecomy were collected at Nanjing Hospital(Nanjing First Hospital), Nanjing Medical University, from January to December 2017. There were 122 male and 61 female patients with age ranging from 42 to 89 years (mean of 68 years). Tumor diameter ranged from 2 to 14 cm (mean 4.5 cm). There were 124 colon cancers and 59 rectal cancers. The number and grade of PDCs in the colorectal adenocarcinoma were evaluated by H-E staining. The overall peritumoural inflammatory reaction was also evaluated. The relationship between PDCs and tumor grades and clinicopathological features and overall peritumoural inflammatory reaction of colorectal adenocarcinoma was analyzed. Results: Of 183 cases of colorectal adenocarcinoma, PDCs were seen in 104 cases (56.8%), of which 36 cases (19.7%) were grade 1, 28 cases (15.3%) were grade 2, and 40 cases (21.9%) were grade 3. PDCs were positively correlated with lymph node metastasis, vascular invasion, degree of differentiation, depth of invasion, and pTNM staging(P<0.05). The detection rate of PDCs in colon cancer was higher than that of rectal cancer(P<0.05). PDCs was unrelated to age, gender, tumor size, and degree of overall peritumoural inflammatory reaction (P>0.05). Among clinicopathological parameters, the grade of PDCs was correlated with lymph node metastasis and vascular invasion (higher than those without lymph node metastasis and vascular invasion, P<0.05); There was a positive correlation between the grade of PDCs and age, tumor differentiation and pTNM staging(P<0.05), and no significant difference between the grade of PDCs and gender, tumor size, tumor location, and depth of invasion was seen (P>0.05). There was no correlation between the grade of PDCs and the degree of overall peritumoural inflammatory reaction (P>0.05). Conclusions: PDC is a histological feature that predicts the aggressiveness of colorectal adenocarcinoma. Evaluation of PDC grade can better predict the biological behavior of colorectal cancer and more accurately guide the treatment and evaluate prognosis.

[Research progress of odontoid fracture in children].

The majority of cervical spine injuries in children occur in the upper cervical spine, of which odontoid fracture is the most common. Odontoid fracture in children is a very insidious injury. Due to the unclear language and incompatible physical examination, the disease is often missed diagnosis. Because the child axis is still in the developmental segment, including 4 synchondrosis and 6 ossification centers, there are obvious anatomical and biological differences between the child odontoid fracture and the adult. Therefore, the choice of treatment is different from that of adults. This article will introduce the development of odontoid in children, and summarize the injury characteristics, clinical classification and treatment of odontoid fracture in children.

[Research progress on meralgia paresthetica after posterior spine surgery].

Meralgia paresthetica (MP) after posterior spine surgery is caused by mechanical compression injury of lateral femoral cutaneous nerve, which presents as numbness or paresthesia in the confined area of the anterolateral thigh. MP after posterior spine surgery is a common postoperative complication, and the incidence is 12.7%-25.5%. Because its clinical manifestations are mild and easy to be ignored, often leading to treatment delays. This article mainly reviews the incidence, risk factors, prevention and treatment of MP after posterior spine surgery.

Label-free rapid identification of tumor cells and blood cells with silver film SERS substrate.

The detection of circulating tumor cells (CTCs) from peripheral blood is considered as great significance for the diagnosis and prognosis of cancer patients. Raman spectroscopy is a highly sensitive optical detection technique that can provide fingerprint molecular identification information. In this paper, the silver film substrate surface-enhanced Raman scattering (SERS) was used to research several tumor cells, immortalized cells, clinical cancer cells isolated from cancer patient's tissue and blood cells. The results display that there is great difference for the nucleic acid characteristic peaks of those cells. The red blood cells have almost none nucleic acid characteristic peak and the SERS signals of white blood cells are only a slight increase. Except for immortalized cells and few tumor cells, the nucleic acid characteristic peaks of some tumor cells have huge enhancement. Nucleic acid characteristic peaks of clinical cancer cells also have greater enhancement. The discriminant model established by the intensity ratio of the nucleic acid characteristic peak 730 cm-1 to the substrate background peak 900 cm-1 shows that some tumor cells and clinical sample cells can be separated from white blood cells, but tumor cells with relatively low-DNA index cannot be differentiated from white blood cells. This study demonstrates that thin-film SERS technology can distinguish between blood cells and some types of tumor cells. This study opens up a new possible method for the detection of CTCs with label-free SERS spectra.

[Clinical characteristics and risk factors in pregnancy with severe community-acquired pneumonia].

Objective: To analyze clinical characteristics of severe community-acquired pneumonia during pregnancy and its outcomes, and to explore the relevant risk factors. Methods: From September 2012 to September 2017, 324 398 pregnancies admitted in 7 tertiary hospitals were included. Clinical data of 33 cases of pregnancies with severe community-acquired pneumonia (severe pneumonia group) and 214 cases of pregnancies with common community-acquired pneumonia (control group) were reviewed retrospectively, including the clinical information, manifestations, laboratory examinations and pregnancy outcomes. Relevant risk factors were analyzed by multivariate logistic regression analysis. Results: (1) General data: pregnancies with severe community-acquired pneumonia accounted for 0.010% (33/324 398) of hospitalized pregnancies, the gestational age of two groups were (28±8) and (23±8) weeks, body mass index were (21.7±2.1) and (25.5±3.4) kg/m(2), rate of low income were 54.5% (18/33) and 31.8% (68/214) , respectively. The differences between two groups were all statistically significant (all P<0.05). No significant differences were found in age, pregnancy and parity times, rate of main pregnant complications such as diabetes and hypertension, educational level, asthma and onset seasons between two groups (all P>0.05). (2) Clinical data: the severe pneumonia group had significantly higher incidence of fever [100.0% (33/33) vs 75.2% (161/214) ], shortness of breath (90.9% vs 16.8%) compared with the control group (all P<0.05) .The median peripheral leukocytes counts were 12.3×10(9)/L and 10.2×10(9)/L, the hemoglobin level were (84±18) and (107±14) g/L,the albumin level were (26±4) and (37±3) g/L, the median serum urea nitrogen level were 3.7 and 2.4 mmol/L, the serum creatinine level were (72±25) and (45±11) µmol/L, respectively in two groups. The differences were all statistically significant (all P<0.05). No significantly statistical differences were found in coagulation indicator and cardiac function between two groups (all P>0.05). (3) Treatments: in severe pneumonia group, 12 patients (36.4%,12/33) needed invasive mechanical ventilation, 9 patients (27.3%,9/33) needed non-invasive mechanical ventilation, average time of mechanical ventilation was (7±4) days;8 patients (24.2%,8/33) with septic shock needed vasoactive drugs. However, there was no patient in control group needing mechanical ventilation and vasoactive drugs. (4) Pregnant outcomes: one patient (3.0%,1/33) died in the severe pneumonia group, while no death occurred in the control group. The hospital stay between two groups were (15.1±4.1) and (7.0±1.9) days, the rates of abortion and stillbirth between two groups were 42.4% (14/33) and 3.3% (7/214) , the rates of premature were 10/19 and 6.3% (13/207) , the rates of cesarean were 15/19 and 43.0% (89/207) , the rates of low birth weight newborn were 17/19 and 14.0% (29/207) , the rates of infected newborn were 15/19 and 10.1% (21/207) , the birth weights were (2 165±681) and (3 102±400) g, respectively. The differences between two groups were all statistically significant (all P<0.05). (5) Multivariate logistic regression analysis demonstrated that anemia, low body mass index, hypoproteinemia were risk factors for severe pneumonia in pregnancy (all P<0.05) . Conclusions: Pregnancy with severe community-acquired pneumonia may be complicated by multiple organ dysfunctions, lead to adverse outcomes. Anemia, malnutrition are risk factors for pregnancy with severe pneumonia. Active and effective treatment may improve its prognosis.

Estimation on Formation Time of Thrombus.

OBJECTIVES: To observe the changes of the formation time of venous thrombus in rats, and to provide new ideas and methods for the estimation on thrombus formation time of the forensic cases died from thrombosis. METHODS: Totally 80 rats were randomly divided into 10 groups （0 h, 3 h, 6 h, 12 h, 1 d, 3 d, 1 week, 2 weeks, 3 weeks and 4 weeks after operation）. A vein thrombosis model was established by the "narrow" method. The processes of thrombosis, organization, recanalization and the features of change on hemosiderin and calcium salt were observed by HE stain, Perls stain and Von Kossa stain. The expression changes of CD61, α-SMA and CD34 were observed by immunohistochemical staining technique. RESULTS: Platelets adhered to the exposed blood vessel intima 3 h after operation, and platelet trabeculae were formed by the repeated accumulation of platelets 1 d after operation. The thrombus organization formed through the fibroblasts from vessel wall that grew into the interior of the thrombus 3 d after operation. Endothelial cells covered the surface of thrombus and then the new blood vessels were reformed, and the vessels were reconstructed. The expression of CD61 upregulated at the stages of the thrombus formation （3 h） and thrombus reformation （4 weeks）, and reached the peak 1 d after thrombus formation. The release of hemosiderin and the initial expression of α-SMA were detected 3 d later. Calcium deposit and expression of CD34 were observed 1 week later. CONCLUSIONS: The hemosiderin, calcium salt, CD61, α-SMA and CD34 show time-dependent changing characteristics, which is expected to provide a reference for the estimation on thrombus formation time of the forensic cases died from thrombosis.

[Perioperative stroke effectively treated by an acute stroke team including anesthesia department: a case report].

Perioperative stroke is cerebral infarction occurring in the perioperative period. The incidence of perioperative stroke in non-cardiac, and non-neurologic surgery is about 0.7%, but the mortality can be as high as 26% to 40%. The outcome of the patients with perioperative stroke can be disastrous. Here we report a case of perioperative ischemic stroke that occurred after surgery of lumbar decompression and pedical screw fixation. A 76-year-old female admitted to our hospital because of lumbar spinal stenosis. Her medical history included hypertension and diabetes for ten years. Her personal history included a smoking history of 60 years by 2 cigarettes per day, not quitting. Her carotid artery ultrasound showed multiple low echo plaques on the right side and multiple high echo plaques on the left side of the carotid artery, but without distinct stenosis. Other examinations and tests showed no distinct abnormality. She went on a lumbar decompression and pedical screw fixation uneventfully. The blood loss was 400 mL and autologous blood transfusion 150 mL. The arterial blood pressure (ABP) maintained during 100-130 mmHg/60-80 mmHg (1 mmHg=0.133 kPa). Sixty minutes after she recovered from general anesthesia, the patient developed symptoms of slurred speech and right limbs weakness. The anesthesio-logist evaluated the patient immediately with National Institute of Health Stroke Scale (NIHSS). The NIHSS score was 11 and a stroke was highly suspected. The acute stroke team was therefore initiated and fast responded. Within 4 h, digital subtraction angiography (DSA) was proceeded, which showed the M1 segment of the left middle cerebral artery was occluded and the local stenosis of her right middle cerebral artery was up to 80%. After the successful embolectomy by Solitaire stent, the left middle cerebral artery reflowed and the forward blood flow was thrombolysis in myocardial infarction (TIMI) grade 3. The patient was discharged after 33 days after the surgery with a NIHSS of 9. Our case provides an example that an acute stroke team that included the department of anesthesiology can be beneficial to the patients' perioperative strokes. During the perioperative period, anesthesiologists should be included into the acute stroke team, because anesthesiologists and anesthesia nurses might be first observers of those early onset strokes. Our case also put forward this thought that a standard peri-operative stroke evaluation tool, like NIHSS, should be discussed and applied to facilitate and accelerate the initiation of perioperative acute stroke team.

[Prenatal diagnosis of 17q12 microdeletion syndrome in fetal renal abnormalities].

Objectives: To analyze 3 cases of 17q12 microdeletion syndrome diagnosed prenatally, and to demonstrate clinical phenotype of the syndrome in prenatal setting. Methods: From January 2013 to July 2017, 1 370 women received invasive prenatal diagnosis and chromosome microarray analysis (CMA) in Peking Union Medical College Hospital. Among them, 3 fetuses were diagnosed as 17q12 microdeletion syndrome. All 3 cases were low-risk pregnancies. Abnormal structures in fetal kidney were found in all 3 cases, including 1 case of multiple renal cysts, 2 cases of bilateral hyperechogenic kidneys. These women accepted invasive prenatal diagnosis followed by karyotyping, parental fluorescence in situ hybridization or CMA validation. Results: The second and third trimester ultrasound showed that all 3 fetuses had bilateral renal structural abnormalities, including hyperechogenic kidney, multiple cysts and renal pelvis dilatation. The karyotyping of the 3 fetuses were normal. CMA examination showed that each case had 1.4-1.6 Mb deletion in 17q12 region. Two cases were de novo deletion and 1 case was inherited from the mother who had mild symptoms. The 3 women decided to terminate pregnancies after genetic counseling. Conclusion: 17q12 microdeletion syndrome is a recurrent chromosome microdeletion syndrome, and the unique phenotype in prenatal setting is the abnormal structure of bilateral kidneys. A few cases of 17q12 microdeletion syndrome even inherited normally phenotypical parents, and prenatal genetic counseling of 17q12 microdeletion syndrome is relatively difficult.

[Prognostic significance of tumor deposits in patients with stage â ¢ colon cancer].

Objective: To investigate the prognostic significance of tumor deposits (TD) in patients with stage Ⅲ colorectal cancer who underwent radical resection. Methods: We retrospectively analyzed the clinicopathological data, and the impact of tumor deposits on the disease free survival (DFS) and overall survival (OS) in 206 patients with stage Ⅲ colorectal cancer using Kaplan-Meier and Cox regression analyses. Results: Among the 206 patients with stage Ⅲ colorectal cancer, there were 112 (54.4%) tumor deposit-positive (TD+ ) cases and 94 (45.6%) tumor deposit-negative (TD-) cases. The TD- and TD+ cases accounted for 10.6% and 21.4% of patients aged <50 years, 22.3% and 36.6% of patients with poorly differentiated carcinoma, and 16% and 29.5% of patients with neural invasion, respectively (P<0.05 for all). In addition, the neural invasion was more often found along with the increasing number of TD. However, multivariate analysis showed that TD was not significantly associated with DFS and OS of the patients. Among the patients with N1 stage cancer, the 3-year DFS rates of TD+ and TD- groups were 23.0% and 58.8%, and the median disease-free survival durations were 26 months and 43 months, respectively (P=0.002). Meanwhile, in the TD- group, the 3-year DFS rates in the patients with stage N1a~1b without tumor deposit and N1c cancer were 63.5% and 21.2%, and the median disease-free survival durations were 41 and 25 months, respectively (P=0.004). Conclusions: Tumor deposit is an independent unfavorable prognostic factor affecting the DFS. The number of tumor deposits may be an important factor affecting prognosis in stage Ⅲ colorectal cancer patients. Tumor deposits after radical resection are associated with neural invasion, and can be used as an independent factor of poor prognostic evaluation in patients with stage N1 colorectal cancer.

[The clinical analysis of instrumentation failure after corrective surgery for thoracolumbar focal kyphosis].

OBJECTIVE: To analyze the etiology of instrumentation failure after corrective surgery for thoracolumbar focal kyphosis, and make suggestion for treatment. METHODS: Retrospective study for 8 patients with thoracolumbar focal kyphosis who underwent surgical treatment and suffered instrumentation failure from June 2005 to December 2011 was made. The surgical procedures included pedicle subtraction osteotomy (PSO), anterior opening-posterior closing osteotomy and correction (AOPC), and posterior vertebral column resection (VCR). The reasons of instrumentation failure were analyzed and revision surgeries were performed. RESULTS: The incidence of instrumentation failure was 6.3%. The average occurrence time was 22.5 months after surgery. Except one had failure in 3 months after surgery, all cases happened after 1 year. In this series, there were 5 cases with post-tuberculosis, 2 cases with post-traumatic kyphosis and 1 case with congenital kyphosis. For the surgical procedure, 7 cases underwent VCR and 1 case AOPC. After the instrumentation failure, all cases had back pain, and 3 of them had combined neurological symptoms. The reasons or risk factors of instrumentation failure included non-fusion of bone graft, VCR procedure, sink of the titanium mesh, insufficiency of anchor sites, and more severe kyphosis. All the 8 cases were treated with revision surgery and got good results. CONCLUSIONS: The instrumentation failure of thoracolumbar focal kyphosis is relatively late occurred, and can develop with various reasons. Positive revision surgery is suggested for the instrumentation failure, and good results can be expected.