ObjectiveTo study the effect of Wulingsan on cerebral edema after intracerebral hemorrhage （ICH） in mice and explore the treatment mechanism. MethodsThe mouse model of ICH was established by injection of collagenase into the caudate nucleus. Mice were randomly assigned into the following groups： sham， ICH， intervention before modeling with low-dose and high-dose （3.69， 11.07 g·kg^-1， respectively） Wulingsan， and intervention after modeling with high-dose Wulingsan. The modified neurological severity score （mNSS） was recorded， and the small animal MRI T2 sequential scanning was performed to measure the volume of cerebral hemorrhage after the modeling of ICH in each group. The Y-maze test， open field test， and Morris water maze test were conducted to evaluate the neurological behaviors of mice in each group. Hematoxylin-eosin staining was employed to observe the pathological changes in the brain tissue. Immunohistochemistry was employed to observe the expression of aquaporin 4 （AQP4）， neuronal nuclei （NeuN）， and glial fibrillary acidic protein （GFAP） in the brain tissue. Western blot was employed to determine the protein levels of AQP4， Claudin-5， and zonula occludens-1 （ZO-1） in the hematoma area. ResultsCompared with the sham group， the ICH group showed increases in the mNSS， the cerebral hemorrhage volume， and the escape latency in the Morris water maze test （P<0.01）， decreases in the times of touching the platform and times of entering the quadrant where the platform was located in the Morris water maze test， and reductions in the spontaneous alternation rate in the Y-maze test and the ratio of distance of center travel to total travel distance in the open field test （P<0.01）. Moreover， pathological changes such as cell disarrangement， cell space enlargement， and cell swelling were observed in the ICH group. Immunohistochemistry results showed that the ICH group had higher proportions of AQP4- and GFAP-positive cells and lower proportion of NeuN-positive cells than the sham group （P<0.01）. Compared with the sham group， the ICH group showed an up-regulated protein level of AQP4 and down-regulated protein levels of Claudin-5 and ZO-1 （P<0.01）. Compared with the ICH group， the intervention with Wulingsan decreased the mNSS， the volume of cerebral hemorrhage， and the escape latency in the Morris water maze test （P<0.05， P<0.01）， while increasing the times of touching the platform and times of entering the quadrant where the platform was located in the Morris water maze test， the spontaneous alternation rate in the Y-maze test， and the ratio of distance of center travel to total travel distance in the open field test （P<0.05， P<0.01）. Furthermore， the intervention with Wulingsan alleviated the pathological changes in the brain tissue after ICH， decreased the proportion of AQP4- and GFAP-positive cells （P<0.01）， increased the proportion of NeuN-positive cells （P<0.01）， down-regulated the protein level of AQP4 （P<0.01）， and up-regulated the protein levels of Claudin-5 and ZO-1 （P<0.01）. ConclusionThe intervention with Wulingsan could reduce the neural function score and the cerebral hemorrhage volume， up-regulate the expression of Claudin-5 and ZO-1， and down-regulate the expression of AQP4 to ameliorate the neurological function defect and cerebral edema after ICH， thereby protecting the brain.

Objective:To investigate the role and mechanism of astrocyte (AS) derived exosomes in protecting brain microvascular endothelial cells (bEnd. 3) from cerebral ischemia reperfusion injury by establishing an oxygen-glucose deprivation/re-oxygenation (OGD/R) model in vitro. Methods:(1) Dual-luciferase reporter gene assay was used to confirm the regulating effect of miR-193b-3p on transient receptor potential cation channel, subfamily M, member 2 (TRPM2). OGD/R model was established by OGD 8 h followed by reoxygenation 24 h in bEnd. 3 cells after being transfected miR-193b-3p mimics/negative sequence (OGD/R+miR-193b-3p mimics group or OGD/R+miR-193b-3p negative sequence group); real-time quantitative PCR (RT-qPCR) was used to detect the miR-193b-3p expression, Western blotting (WB) was used to detect the expressions of TRPM2, cleaved caspase-3, Bax, Bcl-2, ZO-1 and Claudin-5, and flow cytometry was used to detect the apoptosis. (2) AS was extracted from the cerebral cortex of C57BL/6 suckling mice and identified; modeling time was determined by CCK-8 and AS-derived exosomes were extracted by ultracentrifugation from cell supernatant and identified by electron microscopy, particle size analysis, and WB for marker proteins. RT-qPCR was used to detect the miR-193b-3p expression in AS and AS-derived exosomes. Low-expressed miR-193b-3p exosomes were extracted from AS after being transfected miR-193b-3p inhibitory sequence and co-incubated with OGD/R bEnd. 3 cells (group of OGD/R+AS-derived inhibitory sequence exosomes); exosomes were extracted from AS transfecting with miR-193b-3p negative sequence, and co-incubated with OGD/R bEnd.3 cells (group of OGD/R+AS-derived negative sequence exosomes); and normal exosomes were co-incubated with OGD/R bEnd. 3 cells (group of OGD/R+AS-derived exosomes). The miR-193b-3p expression in these 3 groups was detected by RT-qPCR, expressions of TRPM2, cleaved-caspase-3, Bax, Bcl-2, ZO-1, and Claudin-5 were detected by WB, cell apoptosis was detected by flow cytometry, and cell migration ability was detected by scratch test.Results:(1) Dual-luciferase reporter gene assay showed that miR-193b-3p could bind to TRPM2 mRNA. The miR-193b-3p can improve the TRPM2-mediated apoptosis and tight junction reduction in bEnd.3 cells during OGD/R: compared with OGD/R group, OGD/R+miR-193b-3p mimics group had significantly decreased TRPM2, cleaved-caspase-3, and Bax protein expressions, and statistically increased Bcl-2, ZO-1 and Claudin-5 protein expressions ( P<0.05). Flow cytometry further verified the above results: compared with OGD/R group, OGD/R+miR-193b-3p mimics group had significantly decreased cell apoptosis rate (21.34% vs. 13.93%, P<0.05). (2) The extracted exosomes exhibited lipid bilayer cup-like structure with particle size of 126.5 nm (exosome marker proteins: negative Cal and positive CD81 and SG101), indicating successful exosome extraction. After modeling, miR-193b-3p expression in AS and AS-derived exosomes was significantly decreased compared with that in the Control goup ( P<0.05). Compared with the OGD/R group, the group of OGD/R+AS-derived exosomes and group of OGD/R+AS-derived negative sequence exosomes had significantly increased miR-193b-3p expression, statistically decreased TRPM2, cleaved-caspase-3 and Bax protein expressions, and significantly increased ZO-1, Claudin-5 and Bcl-2 expressions ( P<0.05); while compared with those in the OGD/R group, no significant changes in the above protein expressions in group of OGD/R+AS-derived inhibitory sequence exosomes were noted ( P>0.05). Compared with the OGD/R group (18.22%), group of OGD/R+AS-derived exosomes and group of OGD/R+AS-derived negative sequence exosomes had significantly decreased apoptosis rate (14.09% and 13.79%, P<0.05), while group of OGD/R+AS-derived inhibitory sequence exosomes had no significant change (18.41%, P>0.05). Compared with the OGD/R group (13.55%), group of OGD/R+AS-derived exosomes and group of OGD/R+AS-derived negative sequence exosomes had significantly increased migration ability (43.01% and 40.59%, P<0.05), while group of OGD/R+AS-derived inhibitory sequence exosomes had no significant change (16.26%, P>0.05). Conclusion:AS-derived exosomes can suppress TRPM2 protein expression in brain microvascular endothelial cells distantly by miR-193b-3p to improve the brain microvascular endothelial cell injury caused by OGD/R, and then improve OGD/R injury.

Objective:To analyze whether patients with myasthenia gravis (MG) have cognitive impairment and changes of brain structure, and explore the possible mechanisms of cognitive impairment in MG patients from the perspective of brain structure.Methods:Twenty-eight patients with MG admitted to Department of Neurology, First Affiliated Hospital of Soochow University from July 2019 to December 2021 were selected as MG group, and 30 family members from MG patients or healthy subjects who underwent physical examination in Physical Examination Center during the same period were selected as healthy control group. Neuropsychological test was used to evaluate the cognitive function. VBM was used to analyze the changes of brain structure on structural MRI (sMRI). Correlations of gray matter volumes of different brain regions with cognitive function between the two groups were analyzed.Results:Compared with the healthy control group, the MG group had significantly decreased scores of Mini-mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), Clock Painting Test (CDT), and Verbal Fluency Test (VFT), and significantly decreased Rey Auditory Verbal Learning Test (RAVLT) immediate memory and delayed memory scores, while statistically increased time consuming in Making Track Test Part A (TMT-A), and Hamilton Depression Scale (HAMD) and Hamilton Anxiety Scale (HAMA) scores ( P<0.05). Compared with healthy control group, MG group had significantly decreased gray matter volumes of the left orbital superior frontal gyrus, right orbital middle frontal gyrus, right triangular inferior frontal gyrus, left insula, left middle frontal gyrus, right superior limbic gyrus, right anterior cingulate gyrus, right lateral cingulate gyrus, left medial cingulate gyrus, left lateral cingulate gyrus, left medial superior frontal gyrus, and left dorsalateral superior frontal gyrus ( P<0.05). Correlation analysis showed that gray matter volume in the left insula was negatively correlated with time consuming in Stroop Color-Word Test-A ( r=-0.407, P=0.035). Conclusion:Patients with MG may have cognitive decline and gray matter cortical atrophy of some brain regions, and brain areas with gray matter cortical atrophy correspond to areas of cognitive impairment.

Objective: To analyze the association between the occurrence of menarche and the exposure to artificial light at night (ALAN) in female Han nationality girls aged 9-18 in China, so as to provide a reference for the research on the disturbance mechanism of ALAN on puberty, as well as provide a scientific evidence for controlling ALAN and making public sanitary policy. Methods: A total of 74 483 Han girls aged 9-18 with complete records of menarche, height, and weight were included in the 2014 Chinese National Surveys on Students Constitution and Health. Pearson correlation was used to explore the correlation between the incidence of menarche and the exposure to artificial light at night in 11-year-old Han girls, and the third quantiles of the area covered by outdoor night lights (0.16, 0.60) and the third quantiles of the average night light intensity [0.97, 5.91 nw/(sr ·cm) 2] were used respectively to divide the study subjects into 3 groups, and the multifactor Logistic regression model was used to analyze the effect of artificial light at night exposure on menarche. Results: In 2014, the median age of menarche of Chinese Han girls was 12.18 years old, and the incidence of menarche in the 11-year-old group was 26.31%. After controlling for body mass index, local population and the logarithmic values of per capita GDP, the incidence of menarche of 11-year-old Han girls was positively correlated with the local outdoor average night light coverage area ratio and the night average light intensity logarithmic adjustment value( r =0.29, 0.30, P <0.05).The higher the local outdoor night light coverage area ratio ( OR =1.28, 1.52) and the average night light intensity ( OR =1.47, 1.58), the higher the incidence of menarche in 11-year-old Han girls( P <0.05). Conclusion There is a positive correlation between artificial light at night exposure and the menarche rate of 11-year-old Han girls. Research on artificial light at night exposure may provide scientific evidence on puberty onset and its effects on population health.

Lower extremity deep vein thrombosis (DVT) is one of the main complications in patients with traumatic fractures, and for severe patients, the DVT can even affect arterial blood supply, resulting in insufficient limb blood supply. If the thrombus breaks off, pulmonary embolism may occur, with a high mortality. The treatment and rehabilitation strategies of thrombosis in patients with lower extremity fractures have its particularity. DVT in traumatic fractures patients has attracted extensive attention and been largely studied, and the measures for prevention and treatment of DVT are constantly developing. In recent years, a series of thrombosis prevention and treatment guidelines have been updated at home and abroad, but there are still many doubts about the prevention and treatment of DVT in patients with different traumatic fractures. Accordingly, on the basis of summarizing the latest evidence-based medical evidence at home and abroad and the clinical experience of the majority of experts, the authors summarize the clinical treatment and prevention protocols for DVT in patients with traumatic fractures, and make this consensus on the examination and assessment, treatment, prevention and preventive measures for DVT in patients with different fractures so as to provide a practicable approach suitable for China ′s national conditions and improve the prognosis and the life quality of patients.

Objective:To analyze the application effect of discharge planning based on enhanced recovery after surgery (ERAS) in patients with osteoporotic thoracolumbar fracture (OTLF).Methods:A retrospective cohort analysis was made on clinical information of 230 OTLF patients treated in Honghui Hospital of Xi′an Jiaotong University from January to December 2020, including 44 males and 186 females, aged 53-92 years [(72.0±9.9)years]. A total of 115 patients receiving conventional nursing intervention from January to June 2020 were enrolled in regular nursing group and 115 patients receiving discharge planning intervention based on ERAS from July to December 2020 were enrolled in discharge planning group. The length of hospital stay, readiness for hospital discharge scale (RHDS) at 4 hours before discharge, caregiver preparedness scale (CPS) on admission and at 4 hours before discharge, discharge rate before 12∶00, Chinese osteoporosis quality of life short questionnaire (COQOL) on admission and at 6 months after surgery, and re-fracture rate were compared in the two groups.Results:The patients were followed up for 6 months, except for 3 patients lost to follow up in discharge planning group and 4 patients in regular nursing group. The length of hospital stay was (2.8±0.6)days in discharge planning group and (2.6±0.7)days in regular nursing group ( P>0.05). The RHDS in discharge planning group was significantly greater at 4 hours before discharge when compared with regular nursing group [(103.0±8.3)points vs. (95.3±9.5)points] ( P<0.01). The two groups had no significant difference in CPS at admittance ( P>0.05), but a significantly greater CPS was found in discharge planning group at 4 hours before discharge when compared with regular nursing group [(28.9±3.5)points vs. (24.3±4.8)points] ( P<0.01). The discharge rate before 12∶00 in discharge planning group was significantly higher when compared with regular nursing group [27.7%(31/115) vs. 15.3%(17/115)] ( P<0.05). The COQOL was similar at admittance between the two groups ( P>0.05), but a significantly lower score was found in discharge planning group than that in regular nursing group [(21.6±6.2)points vs. (26.6±6.9)points] ( P<0.01). A significantly lower re-fracture rate was found in discharge planning group at 6 months after surgery when compared with regular nursing group [4.5%(5/112) vs. 12.6%(14/111)] ( P<0.05). Conclusion:For OTLF patients, discharge planning based on ERAS is superior to regular nursing in improving the readiness for hospital discharge, caregiver preparedness, quality of life and management of beds, and lowering re-fracture rate.

Objective:To evaluate the effect of goal-directed fluid therapy (GDFT) based on permissive high stroke volume variation (SVV) guidance on residual liver function in elderly patients undergoing laparoscopic hepatectomy.Methods:A total of 100 elderly patients of either sex, aged 65-80 yr, with body mass index of 18.5-24.9 kg/m 2, of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with Child-Pugh grade A or B, scheduled for elective laparoscopic hepatectomy, were divided into 2 groups ( n=50 each) by the random number table method: SVV-guided GDFT group (group SG) and CVP-guided fluid replacement group (group C). Intraoperative fluid management was divided into 2 stages.The first stage was from the start of surgery to the completion of liver resection, the SVV was maintained at 13%-20% in group SG, and the low CVP was maintained at 0-5 cmH 2O in group C. The second stage was from completion of liver resection to the end of the operation, SVV was maintained at 9%-13%, additional hydroxyethyl starch 3 ml/kg was given or repeatedly administered when SVV>13% (for 5 min) or when the response to previous fluid replacement was positive (SVV increased by more than 10%), and the infusion rate was slowed down when the SVV was 9%-13% in group SG, and CVP was maintained at 5-12 cmH 2O in group C. Mean arterial pressure and heart rate were recorded on admission to the operating room, at skin incision (T 1), at the start of liver resection (T 2), at completion of liver resection (T 3) and at the end of operation (T 4). The operation time, intraoperative blood loss, transfusion volume, urine volume and levels of serum lactic acid before operation and at the end of operation were recorded.Blood samples from the median cubital vein were collected at T 0-4 to measure blood glucose and cortisol concentrations.The concentrations of serum aspartate aminotransferase, alanine aminotransferase, total bilirubin and albumin were measured before operation, at 1, 3 and 5 days after operation, and prothrombin time, activated partial thromboplastin time, thrombin time and Fib were recorded.The concentrations of serum interleukin-6, tumor necrosis factor-alpha and C-reactive protein were measured by enzyme-linked immunosorbent assay before operation and at the end of operation, and the postoperative complications and length of hospital stay were recorded. Results:Compared with group C, mean arterial pressure and heart rate were significantly decreased at T 2, 3, blood loss was reduced, transfusion volume and urine volume were increased, prothrombin time and activated partial thromboplastin time were shortened at the end of operation, serum concentrations of interleukin-6 and lactic acid and concentrations of aspartate aminotransferase and alanine aminotransferase in serum at 5 days after operation were decreased, and the length of hospital stay was shortened in group SG ( P<0.05). Conclusions:GDFT based on permissive high SVV guidance can improve residual liver function in elderly patients undergoing laparoscopic hepatectomy.

Objective:To analyze the clinical manifestation, laboratory examination, treatment and prognosis of congenital factor Ⅺ (FⅪ) deficiency.Methods:The clinical data of 80 patients with congenital FⅪ deficiency in our hospital from September 2006 to October 2020 were analyzed retrospectively.Results:Among the 80 patients, there were 33 males (41.3%) and 47 females (58.8%) , with a median age of 32 (2-66) years. Twenty-eight cases (35.0%) had bleeding events, including 11 cases of spontaneous bleeding (13.8%) , 9 cases of ecchymosis or bleeding after skin trauma (11.3%) , 9 cases of postoperative bleeding (11.3%) . Among the female patients, there were 11 cases of menorrhagia (23.4%) and 1 case of bleeding after vaginal delivery (2.1%) . Laboratory examination were characterized by prolonged activated partial thromboplastin time (APTT) , normal prothrombin time (PT) , and decreased FⅪ activity (FⅪ∶C) . Nine patients (11.3%) were tested for FⅪ gene (F11) with 11 mutations. Twenty-seven patients (33.8%) received fresh frozen plasma (FFP) treatment, 15 patients (18.8%) were received for prophylaxis with no bleeding occurred during and after operation.Conclusion:Most patients with congenital FⅪ deficiency have no or mild bleeding symptoms. There was no significant correlation between FⅪ∶C and the severity of bleeding symptoms, and there was a well consistency between FⅪ∶C and F11 homozygous or heterozygous mutation type. Prophylactic infusion of FFP can effectively reduce the risk of operative bleeding.

Objective:To analyze the clinical phenotype and molecular pathogenesis of nine patients with hereditary factor Ⅴ (FⅤ) deficiency.Methods:Nine patients with hereditary FⅤ deficiency who were admitted to the Institute of Hematology and Blood Diseases Hospital from April 1999 to September 2019 were analyzed. The activated partial thromboplastin time, prothrombin time, and FⅤ procoagulant activity (FⅤ∶C) were measured for phenotypic diagnosis. High-throughput sequencing was employed for the F5 gene mutation screening, Sanger sequencing was adopted to confirm candidate variants and parental carrying status, Swiss-model was used for three-dimensional structure analysis, and ClustalX v.2.1 was used for homologous analysis.Results:The FⅤ∶C of the nine patients ranged from 0.1 to 10.6. Among them, eight had a hemorrhage history, with kin/mucosal bleeding as the most common symptom (three cases, 37.5%) , whereas one case had no bleeding symptom. There were five homozygotes and four compound heterozygotes. A total of 12 pathogenic or likely pathogenic mutations were detected, of which c.6100C>A/p.Pro2034Thr, c.6575T>C/p.Phe2192Ser, c.1600_1601delinsTG/p. Gln534*, c.4713C>A/p.Tyr1571*, and c.952+5G>C were reported for the first time.Conclusion:The newly discovered gene mutations enriched the F5 gene mutation spectrum associated with hereditary FⅤ deficiency. High-throughput sequencing could be an effective method to detect F5 gene mutations.

Objective:To investigate the clinical type and gene mutations, clinical manifestations, laboratory tests, diagnosis, and fibrinogen replacement therapy of congenital fibrinogen disorders.Methods:Clinical data of 146 patients with congenital fibrinogen disorders diagnosed from April 2000 to November 2020 were retrospectively analyzed.Results:Among the 146 patients, 61 (41.8%) men and 85 (58.2%) women had a median age of 33.5 years at the time of consultation. 34 patients (34.7%) were found to suffer from the disease due to bleeding symptoms, 33 patients (33.7%) due to preoperative examination. 55 patients (56.1%) had at least one bleeding symptom, and 42 patients (42.9%) had no bleeding symptoms. There is a negative correlation between fibrinogen activity concentration and bleeding ISTH-BAT score (rs=-0.412, P=0.001) . A total of 34 gene mutations were detected in 56 patients, of which 84.1% were missense mutations, and 16 new mutations were found. FGA Exon2 and FGG Exon8 mutations accounted for 71.4% of all mutation sites. Patients with afibrinogenemia were younger, with a median age of 2 (1-12) years, an ISTH-BAT score of 4, and patients with dysfibrinogenemia had significantly longer thrombin time (TT) , with a median of 28.5 (19.2-36.6) s. The 1 hour in vivo recovery (IVR) after fibrinogen infusion was (127.19±44.03) %, and the 24 hour IVR was (101.78±43.98) %. In addition to the obvious increase in the concentration of fibrinogen activity, the TT and the prothrombin time (PT) both decreased significantly, and the TT decreased more significantly, with an average decrease of 15.2% compared to the baseline after 24 hours of infusion. Conclusion:Most patients with congenital fibrinogen disorders have mild or no bleeding symptoms. Patients with afibrinogenemia have more severe symptoms. There is a negative correlation between the fibrinogen and the degree of bleeding. Genetic testing is helpful for the diagnosis of disease classification. FIB∶C/FIB∶Ag<0.7 can be used as a basis for clinical diagnosis. The TT can be used as the basis for the diagnosis of dysfibrinogenemia and the effectiveness of fibrinogen infusion.