BACKGROUND:Shujin Jiannao Prescription is an empirical formula for the treatment of cerebral palsy in Dongzhimen Hospital,Beijing University of Chinese Medicine,with clear clinical efficacy,but the specific mechanism needs to be elucidated. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into a normal group(n=12)and a model group(n=52).An animal model was established by the Rice-Vannucci method.After successful modeling,52 model rats were randomly divided into control model group(n=12),minocycline group,and the low-,medium-,and high-dose groups of the Shujin Jiannao Prescription(n=10 per group).Rats in the minocycline group were given 40 mg/kg·d minocycline by gavage;rats in the low-,medium,and high-dose groups were given 4,8,and 16 g/kg·d Shujin Jiannao Prescription granules by gavage,respectively;and rats in the normal group and control model group were given an equal dose of normal saline by gavage.Medication in each group was given once a day for 1 week.The rats in each group were evaluated behaviorally using suspension test,abnormal involuntary movement score,and Bederson score.The pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining.The levels of tumor necrosis factor α,interleukin 1β,and interleukin 10 in the cerebral cortex were determined using ELISA.The positive expressions of Janus kinase 2(JAK2),phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)in the cerebral cortex were detected using immunohistochemistry.The protein expression levels of JAK2,p-JAK2,and p-STAT3 were detected using western blot. RESULTS AND CONCLUSION:Compared with the normal group,the suspension test score and involuntary movement score were decreased in the control model group(P＜0.01 or P＜0.05).The pathological results showed structural disruption of nerve cells,formation of large numbers of vacuoles,cell swelling,and increased intercellular space in the control model group.In addition,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were significantly increased(P＜0.01),the expression of interleukin 10 was decreased(P＜0.05),and the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly increased(P＜0.01)in the control model group compared with the normal group.Compared with the model group,minocycline and Shujin Jiannao Prescription at each dose could improve the behavioral indexes of rats(P＜0.01 or P＜0.05)and ischemic-hypoxic pathological changes were attenuated,with only a small amount of necrotic nerve cells and a few vacuoles,and reduced intercellular space.Moreover,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were decreased in each drug group compared with the control model group(P＜0.05),while the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly decreased(P＜0.01).The most obvious improvement was observed in the high-dose Shujin Jiannao Prescription group.To conclude,Shujin Jiannao Prescription can inhibit inflammation in the cerebral cortex of rats with hypoxic-ischemic brain injury.The mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway.

BACKGROUND:Perinatal hypoxic-ischemic brain injury is one of the most common causes of cerebral palsy.Shujin Jiannao Prescription is an experienced formula for treating cerebral palsy and improving blood supply to the brain developed by the Dongzhimen Hospital,Beijing University of Chinese Medicine. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating hypoxic-ischemic cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into six groups.There were 12 rats in each of the control and model groups as well as 10 animals in each of the minocycline group,and the low-,medium-,and high-dose groups of Shujin Jiannao Prescription.The neonatal rat ischemic-hypoxic cerebral palsy model was established in all groups except for the control group.After successful modeling,rats in each drug group were respectively gavaged with minocycline and Shujin Jiannao Prescription at a dose of 4,8,and 16 g/kg per day for 1 week.Body mass of rats was measured and behavioral changes were detected before and after drug administration.Hematoxylin-eosin staining was used to observe the histomorphology of hippocampal CA1 region of rat brain tissue,and immunohistochemistry and western blot were used to detect the expression levels of Bcl-2,Bax,and Caspase-3 in the brain tissue of rats. RESULTS AND CONCLUSION:Compared with the model group,medium-and high-dose Shujin Jiannao Prescription significantly increased the body mass of rats(P<0.05).Compared with the model group,minocycline effectively prolonged the suspension time of ischemic-hypoxic cerebral palsy rats(P<0.05),while medium-and high-dose Shujin Jiannao Prescription significantly prolonged the suspension time,shortened the inclined plane test time,and increased the Longa score of rats(P<0.05).The pathological results showed that after drug intervention,only a small number of neuronal cells in the brain tissue of rats were necrotic,the cells were more neatly arranged,the cell structure was more complete,and only part of the cell nuclei became smaller.Compared with the model group,minocycline and medium-and high-dose Shujin Jiannao Prescription reduced the expression of Bax Caspase-3(P<0.05),medium-and high-dose Shujin Jiannao Prescription increased the expression of Bcl-2(P<0.05),and Bcl-2/Bax protein expression was increased in minocycline and three Shujin Jiannao Prescription groups(P<0.05).In addition,the protein expression was increased in a dose-dependent manner after intervention with Shujin Jiannao Prescription,and there was no significant difference between the minocycline and three Shujin Jiannao Prescription groups(P>0.05).To conclude,the mechanism by which Shujin Jiannao Prescription treats ischemic-hypoxic cerebral palsy in rats may be to enhance the expression of anti-apoptotic protein Bcl-2,inhibit the expression of pro-apoptotic protein Bax,and reduce the expression of Caspase-3,ultimately inhibiting the apoptosis of hippocampal neuronal cells in rats with cerebral palsy.Within a certain range,the higher dose of Shujin Jiannao Prescription indicates the better therapeutic effect,and the high-dose Shujin Jiannao Prescription is as effective as minocycline.

COVID-19 associated Guillain-Barré syndrome (GBS) caused by peripheral nerve damage after SARS-CoV-2 infection is one of the most common COVID-19 related nervous system inflammatory diseases, with high incidence of respiratory failure and mortality. Positive SARS-CoV-2 RNA in cerebrospinal fluid of COVID-19 associated GBS patients has been rarely reported. This paper reports 4 patients with COVID-19 associated GBS in China who developed neurological symptoms 4-15 days after fever and were confirmed SARS-CoV-2 infection. All patients presented with progressive weakness of both lower limbs, 3 patients with autonomic dysfunction such as defecation and urination disorders, and 1 patient with polycranial neuritis and Miller-Fisher syndrome such as bilateral facial palsy, dysphagia, diplopia and ataxia. Nerve conduction velocity and F wave were abnormal in 3 patients, and motor conduction pathway was abnormal in 1 patient. Anti-ganglioside antibodies were tested in 3 patients, and GD1a-IgG was positive in 1 patient. All 4 patients underwent metagenomic next-generation sequencing examination in blood and cerebrospinal fluid. SARS-CoV-2 RNA was positive in blood and cerebrospinal fluid of 3 patients, and SARS-CoV-2 RNA was positive in cerebrospinal fluid of 1 patient.

Objective:To analyze the F10 gene mutations in a Chinese pedigree affected with the deficiency of the hereditary coagulation factor X (FX), resulting from a new deletion mutation, and to study the associated molecular pathogenesis.Methods:Next generation sequencing (NGS) was performed to screen the genetic mutations in the proband which were then verified by Sanger sequencing. The FX activity (FX∶C) of probands and their family members was detected using the blood clotting method, and the mutation sites of the family members were analyzed using Sanger sequencing. The pathogenicity of the mutation site was predicted by using the online bioinformatics software, Mutation Taster. The SWISS-MODEL software was used for stimulating the three-dimensional models of the wild-type and mutant proteins for analyzing the influence of the mutation site on the structure and function of the proteins, and for analyzing the difference between the catalytic residues of the wild-type and the mutant proteins. The level of the F10 gene mRNA was quantitatively analyzed by qRT-PCR (quantitative reverse transcription polymerase chain reaction) method by constructing plasmids, transfecting human embryonic kidney 293T cells (HEK 293T), and analyzing the splicing of the mutated site by RT-PCR method. The levels of FⅩ∶Ag in cell lysates and cell culture media (both inside and outside the cells) were detected by the ELISA (enzyme linked immunosorbent assay) method.Results:A medium-grade factor X deficiency with a 36.42% FⅩ∶C ratio was detected in the proband by the coagulation method. NGS analysis demonstrated a heterozygous deletion mutation in exon 8:c.902_919del (p.Ala301_Glu306del) in the proband. Sanger sequencing analysis indicated that some members of the family (mother and grandfather) were also carriers of the corresponding deletion mutation. Online bioinformatics software predicted the pathogenic nature of the c.902_919del mutation, with a pathogenic score of 0.999. The 3D protein structure model analysis indicated that the c.902_919del mutation resulted in the disappearance of a segment of β-fold in the protein structure, thereby shortening the preceding segment of the β-fold and a subsequent loss of hydrogen bonds between adjacent amino acids with no significant difference in the side chain conformation of the key catalytic residues compared to the wild-type. mRNA splicing analysis indicated the absence of alternative splicing changes in the mutation, and qRT-PCR results indicated the absence of a statistically significant difference between the mRNA levels of F10 gene and wild-type mRNA in cells expressing c.902_919del mutant. The ELISA results indicated that there was no statistically significant difference in the FX∶Ag levels of the mutant cell culture medium and the lysate.Conclusions:In this pedigree, the heterozygous mutation in exon 8 of F10 gene (c.902_919del, p.Ala301_Glu306del) caused the hereditary factor Ⅹ deficiency.

A good social network can ensure the safety of women during pregnancy and reduce the incidence of adverse pregnancy outcomes. However, there is currently relatively little research on the overall social network of pregnant women, and the influencing factors of the overall social network of pregnant women are not yet clear. This paper reviews the current situation, influencing factors, and measurement tools of the social network in pregnant women, aiming to improve their social network, help them access high-quality nursing resources, ensure the health of pregnant women and infants, and decrease the incidence of adverse pregnancy outcomes.

Purpose To study the association between ultrasonography signs of midurethral sling(MUS)and postoperative bladder neck mobility,and urethral segmental mobility,to explore ultrasound parameters that measure the biomechanical effects of MUS and to analyze the relationship between them and the clinical outcomes.Materials and Methods This was a retrospective analysis of the clinical material and ultrasound imaging data of the patients who underwent MUS surgery and had postoperative clinic follow-up in the Second Xiangya Hospital,Central South University,from September 2017 to July 2022.According to the surgical outcome,all patients were divided into three groups:stress urinary incontinence(SUI)cure group,SUI recurrence group and postoperative voiding dysfunction(VD)group.Bladder neck mobility,urethral segmental mobility,MUS position,and sling-pubic gap(SPG)during maximal Valsalva manoeuvre were measured by pelvic floor ultrasound.Ultrasound results among the three groups were compared,respectively.The relationships between ultrasound signs of the sling(MUS position and SPG),bladder neck and urethral mobility,and the surgical outcomes were analyzed,respectively.Results A total of 117 women had valid data.The median follow-up interval was 10(6,18)months.On clinical examination and diagnosis,44 women(37.6％)had cured SUI,46(39.3％)had recurrence SUI,and 27(23.1％)had postoperative VD.The mean SPG of the 117 slings was(12.0±3.5)mm(range 4.7 to 23.0 mm),and the mean position of the MUS was the 53％(range 33％-75％).There was no significant difference in MUS position and SPG between the SUI cured group and the postoperative VD group(P＞0.05).The SUI recurrence group had farther MUS position[(56±11)％vs.(49±10)％,P=0.003]relative to the bladder neck and wider SPG[(13.9±3.7)mm vs.(11.2±2.7)mm,P＜0.001]than SUI cure group.No significant correlation was found between the ultrasound signs of MUS(MUS position and SPG)and bladder neck mobility(r=-0.138-0.205,all P≥0.05).MUS position and SPG were correlated with midurethral mobility(MUS position vs.point 2 and 3,r=0.322,0.322,both P＜0.01;SPG vss.point 3 to 6,r=0.288-0.434,all P＜0.01):the closer the MUS position was relative to the distal urethra,the higher the midurethral mobility.The wider the SPG,the higher the midurethral mobility.Logistic regression showed a positive correlation between SPG and SUI recurrence with an odds ratio(OR)of 1.401(95％CI 1.189-1.652,P＜0.001),and a negative correlation with postoperative VD with an OR of 0.755(95％CI 0.627-0.909,P=0.003).Conclusion SPG during the Valsalva manoeuvre can be used to measure the tightness of MUS.The larger the measured value of SPG,with the looser the MUS,the greater the likelihood of postoperative SUI recurrence,and the lower the risk of postoperative VD.

Objective:To study the agreements between transperineal ultrasound (TPUS) and endoanal ultrasound in assessing obstetrics anal sphincter injury (OASI), and to analyse the diagnostic efficacy of OASI in predicting AI relationship between OASI and anal incontinence (AI).Methods:A total of 217 women were prospectively recruited from the clinic in the Second Xiangya Hospital of Central South University from January 2021 to May 2022. Symptoms of AI were determined using the St Mark′s Incontinence Score (SMIS). TPUS and EAUS were performed by the same operator with the same machine on every participant for detecting OASI: OASI grades 3a, 3b, 3c, and 4 were performed according to the extent of the injuries in the anal sphincter complex. The angle of the defect in the external anal sphincter (EAS) was measured. A "significant EAS defect" was diagnosed as a defect affecting at least 2/3 of the length of the EAS with a defect angle of ≥30° in each slice.Ultrasound findings were compared between the two methods. The diagnostic efficacy of "ultrasound OASI" in predicting AI was analysed by logistic regression.Results:Of 217 women, twenty-eight (12.9%) suffered from AI with SMIS ranging from 5~20(11.9±4.5). On TPUS, 79 (36.4%) cases were suspected of OASI, that was 50 OASI 3a, 13 OASI 3b, and 16 OASI 3c/4. On EAUS, 78 (35.9%) cases were suspected of OASI that was 23 OASI 3a, 22 OASI 3b, 15 OASI 3c, and 18 OASI 4. Twenty-four "significant EAS defects" were diagnosed by TPUS and twenty-eight by EAUS, TPUS had excellent agreement with EAUS (weighted Kappa=0.91, P<0.001). Logistic regression analysis showed that "ultrasound OASI" was associated with AI symptoms. ROC curve analysis showed that the area under the curve (AUC) was 0.92, 0.87, 0.89, 0.92 for TPUS OASI 3b+ , EAUS OASI 3b+ , TPUS "Significant EAS defect" , and EAUS "Significant EAS defect" for predicting AI, respectively. Conclusions:TPUS has good agreement with EAUS in detecting OASI. OASI 3b+ and "significant EAS defect" on TPUS and EAUS had good performance in predicting AI symptoms.

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
Pregnancy ; Female ; Animals ; Mice ; Tetraploidy ; Blastocyst ; Embryo, Mammalian ; Cell Differentiation ; Embryonic Development

Objective:To evaluate the new model of group screening combined with opportunistic screening for the diagnosis and treatment of gastric cancer.Methods:Group screening combined with opportunistic screening was used for gastric cancer screening. (1) Group screening. Cluster sampling was used to screen gastric cancer by endoscopy in high-risk population (aged 40-<70 years) of rural residents in Weihai from July 2017 to December 2020, and biopsy was obtained for histopathology if necessary. Main collection parameters included the detection rate of advanced gastric cancer, early gastric cancer and high-grade intraepithelial neoplasia (HGIN). (2) Opportunistic screening. The changes of the detection rates of early gastric cancer in opportunistic screening in 2 hospitals in Weihai area were observed during the same period of time.Results:(1) In group screening, from July 2017 to December 2020, the first batch of 8 000 cases of gastric cancer screening were completed. The cases of advanced gastric cancer, early gastric cancer and HGIN were 36, 28, and 62, respectively. The detection rates of gastric cancer and early gastric cancer were 0.80% (64/8 000) and 43.75% (28/64), respectively. The proportion of early gastric cancer+HGIN who received endoscopic submucosal dissection (ESD) was 77.78% (70/90), and the rate of curative resection was 100.00%(70/70). (2) Opportunistic screening: from July 2017 to December 2020, the annual early gastric cancer detection rates in opportunistic screening in Wendeng District Traditional Chinese and Western Medicine Hospital were 16.67% (1/6), 20.00% (3/15), 23.53% (4/17), and 33.33% (6/18) in the consecutive 4 years, respectively. The annual detection rates of early gastric cancer in opportunistic screening in Ru Shan Peoples Hospital were 14.74% (14/95), 23.80% (60/252), 25.49% (65/255), and 24.04% (50/208), respectively. The detection rates of opportunistic screening for early gastric cancer in hospitals in Weihai city increased year by year.Conclusion:In areas with high incidence of gastric cancer, a certain scale of group screening can lead to a wider range of opportunistic screening, resulting in the increase of the detection rate of early gastric cancer. The new model of diagnosis and treatment of gastric cancer is worth recommendation.

It is still at the initial stage for basic-level military doctors to participate in the standardized residency training. While, their professional backgrounds and training objectives are significantly different from those civilian residents, it is necessary to explore the standardized training methods in line with the military career demands for those basic-level military doctors. Based on the actual needs of grass-roots troops and military medicine, this study makes significant progress in the integration of standardized residency training and field surgery training methods, making the basic-level military doctors achieve the goal of field surgery training in the course of clinical practice of residents. Not only the surgical capability of basic-level military doctors, but also the medical support capability in grass-roots troops has been improved. This article summarizes the application methods and experience of field surgery teaching in the standardized residency training of military medical residents at the grass-roots level.