OBJECTIVE To explore the molecular mechanism of Compound Dihuang Granule in treating Parkinson's disease(PD)with yin-deficiency and dynamic wind syndrome by regulating the changes of ubiquitin-proteasome system-related proteins.METHODS SPF male SD rats were randomly selected as normal control group and sham operation group,13 rats in each group.The remaining rats were treated with 6-hydroxydopamine lateral brain injection to damage the substantia nigra to establish PD yin-deficien-cy and dynamic wind syndrome rat model.The established rats were randomly divided into model group,madopar group(150 mg·kg-1),low,medium and high dose groups of Compound Dihuang Granule(1.75,3.5,7 g·kg-1),13 rats in each group.Each group was given drugs by intragastric gavage for 28 d.The general conditions and neurobehavioral manifestations of the rats were observed;the expression of ubiquitin(UB),ubiquitin activating enzyme(UBE1),ubiquitin conjugating enzyme(UBE2A)and ubiquitin carboxyl terminal hydrolase(UCH-L1)positive cells,protein and mRNA expression levels in the damaged lateral stria-tum of the brain of rats were detected by immunohistochemistry,Western blot and qPCR methods;changes of dopamine(DA)content in the damaged lateral striatum of the brain of rats were detected by ELISA.RESULTS There was no significant difference in general conditions between the normal control group and the sham operation group,and there was no significant difference in the number of ro-tations,suspension time and number of moving grids.Compared to the sham-op group,the model group rats showed poor spirit,irrita-bility,limb tremor,slow movement,dark yellow fur,less food intake,reduced body weight;the modeling increased number of rota-tions,decreased hanging time and number of moving grids(P<0.01),significantly decreased expression of UB,UBE1,UBE2A,and UCH-L1 positive cells,mRNA and protein expression levels in the damaged lateral striatum(P<0.01),and significantly decreased DA content(P<0.01).Compared with the model group,the general condition of the rats in the madopar group and Compound Di-huang Granule groups was improved,the number of rotations was reduced,the hanging time and number of moving grids were increased(P<0.05,P<0.01),the expression of UB,UBE1,UBE2A,and UCH-L1 positive cells,mRNA and protein expression levels were increased(P<0.05,P<0.01),and the DA content was increased(P<0.01);behavioral tests and the expression levels of various in-dicators were most significantly improved in the madopar group and the high-dose Compound Dihuang Granule group(P<0.05,P<0.01),and there was no statistical significance between the two groups.CONCLUSION Compound Dihuang Granule may play a role in treating PD yin-deficiency and dynamic wind syndrome by regulating the expression of ubiquitin-proteasome system-related proteins,reducing abnormal protein aggregation,and thus alleviating DA content deficiency,and the high-dose Compound Dihuang Granule has the best effect.

Objective To observe the effects of compound Dihuang Granules on striatum PINK1,Parkin and α-syn in rats with Parkinson disease yin-deficiency and wind movement syndrome;To explore the mechanism of compound Dihuang Granules in the treatment of Parkinson disease yin-deficiency and wind movement syndrome.Methods 6-hydroxydopamine was injected into the substantia nigra of the midbrain to prepare the model of PD yin-deficiency and wind movement syndrome.Totally 65 model rats were randomly divided into model group,Madopar(150 mg/kg)group,and TCM low-,medium-,and high-dosage(1.75,3.5,7 g/kg)groups.Another 26 rats were divided into normal control group and sham-operation group,with 13 rats in each group.Each administration group was given corresponding medication solution by gavage,the normal control group,sham-operation group and the model group were given equivalent volume of normal saline,once a day for 28 consecutive days.The behavioral changes of rats were detected,ELISA and biochemical assay kits were used to detect ATP content and COX Ⅳactivity in striatum,HE staining was used to observe the morphological changes of the striatum,immunohistochemistry staining,Western blot and RT-qPCR were used to detect the expression of PINK1,Parkin,α-syn protein and mRNA in striatal tissue.Results Compared with the normal control group and the sham-operation group,the rats in the model group showed increased number of rotations and pole climbing time,decreased swimming score(P<0.01),the ATP content and COX Ⅳ activity in striatum tissue decreased(P<0.01),with disordered arrangement and reduced number of neurons,cell swelling,partial nuclear shrinkage,formation of vacuoles,and blurred boundaries,the expressions of PINK1,Parkin protein and mRNA in striatal tissue decreased(P<0.01),and the expressions of α-syn protein and mRNA increased(P<0.01).Compared with the model group,the number of rotations and pole climbing time of rats in each medication group decreased,swimming scores increased(P<0.01),ATP content and COX Ⅳ activity in striatal tissue increased(P<0.05,P<0.01),the number of neurons increased,the arrangement tended to be neat,and the morphology improved,the expressions of PINK1,Parkin protein and mRNA in striatal tissue increased(P<0.05,P<0.01),the expressions of α-syn protein and mRNA decreased(P<0.05,P<0.01).The improvement was most significant in the Medopar group and TCM high-dosage group(P<0.05,P<0.01),and there was no significant difference between the two groups(P>0.05),and TCM groups showed dosage dependence(P<0.05,P<0.01).Conclusion Compound Dihuang Granules can improve the behavioral symptoms of Parkinson disease yin-deficiency and wind movement syndrome,possibly by regulating the expressions of PINK1,Parkin and α-syn in the striatum tissue,thereby alleviating mitochondrial dysfunction,protecting dopaminergic neurons,and playing a role in treating Parkinson disease.

Objective To observe the effect of compound Dihuang Granules on the expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver and colonic tissue of Parkinson disease(PD)rats with yin deficiency and wind movement syndrome.Methods 6-Hydroxydopamine was used to inject into the right substantia nigra to prepare PD with yin deficiency and wind movement syndrome rats model.65 rats that successfully modeled were randomly divided into model group,madopar group(madopar 150 mg/kg)and TCM low-,medium-and high-dosage groups(compound Dihuang Granules 1.75,3.5,7 g/kg),with 13 rats in each group;another 13 rats without intervention were selected as the normal control group,and 13 rats were only injected with ascorbic acid solution as sham-operation group.The administration groups were given corresponding drugs by gavage,and the normal control group,sham-operation group and model group were given normal saline by gavage,once a day for 28 consecutive days.The neurobehavioral changes of rats were observed,and the morphology of liver tissue and colonic tissue were observed by HE staining,the protein and mRNA expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver and colonic tissue were detected by immunohistochemical staining,Western blot and RT-PCR,respectively.Results Compared with the normal control group and the sham-operation group,the number of rotation circles of rats in the model group increased,with granular degeneration of liver tissue,loose cytoplasm of cells,slight hepatic steatosis,edema of colonic mucosal and submucosal layers,loose connective tissue,infiltration of a small number of lymphocytes,and significant vascular dilation,the expressions of DJ-1,IP3R,GRP75 and VDAC1 protein and mRNA in liver and colonic tissue were significantly decreased(P<0.01).Compared with the model group,the number of rotation circles of rats in madopar group and TCM groups were significantly decreased,with mild granular degeneration of liver cells,loose cytoplasm,orderly arrangement of hepatic cords,mild edema of the colon,a small amount of lymphocyte and granulocyte infiltration,and a small amount of vascular dilation,the protein and mRNA expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver and colonic tissue significantly increased(P<0.05).The TCM high-dosage group showing the most obvious changes,there was no statistical significance with the madopar group(P>0.05).Conclusion Compound Dihuang Granules may restore endoplasmic reticulum-mitochondria homeostasis,improve gut-liver-brain axis mitochondrial function,protect dopamine neurons,and delay PD by regulating the expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver tissue and colonic tissue.

Objective:To investigate the genetic etiology of fetal conotruncal heart defects (CTDs) and to evaluate the performance of copy number variation sequencing (CNV-seq) and whole exome sequencing (WES) in identifying the genetic etiology.Methods:This retrospective study involved 196 fetuses diagnosed with CTDs by fetal echocardiography in Beijing Anzhen Hospital, Capital Medical University from June 2017 to December 2021. CNV-seq was performed to screen for chromosomal abnormalities [aneuploidy and copy number variations (CNVs)] in the fetuses and their parents, and then WES was performed if CNV-seq was negative. The diagnostic yields of genetic abnormalities [aneuploidy+CNVs+single nucleotide variations (SNVs)] for different types of CTDs were compared using Chi-square test. Results:CNV-seq revealed 54 cases (27.6%, 54/196) with chromosomal abnormalities, including 14 (7.1%, 14/196) aneuploidies, 39 (19.9%, 39/196) CNVs and one aneuploidy complicated by CNVs. Together with another 13 fetuses with pathogenic or likely pathogenic SNVs detected by WES among the rest 142 cases whose CNV-seq results were negative, the total detection rate of genetic abnormalities was 34.2% (67/196). WES increased the diagnostic yield for CTDs by 9.2% (13/142). There was significant difference in the diagnostic yields for different types of CTDs ( χ2=20.31, P=0.002). The diagnostic yield was relatively high for interrupted aortic arch of type B, absent of the pulmonary valve -type of tetralogy of Fallot (9/10 and 8/12), but low for transposition of the great arteries (12.5%, 5/40). Conclusions:CNVs are the common genetic abnormalities in fetal CTDs, and SNVs are also detected in some cases. It is recommended that all fetuses with CTDs should undergo genetic testing. CNV-seq should be used in combination with WES if possible to improve the identification of genetic etiology and provide reference for genetic counseling.

OBJECTIVE: To summarize the clinical features and spectrum of genetic variants in 12 patients with Loeys-Dietz syndrome (LDS), and to explore the correlation between the type of genetic variants and clinical phenotypes. METHODS: Twelve patients suspected for LDS at Beijing Anzhen Hospital Affiliated to Capital Medical University from January 2015 to January 2022 were selected as the study subjects. Clinical data of the patients were collected. Genomic DNA was extracted from peripheral blood samples and subjected to genetic testing. Pathogenicity of candidate variants was analyzed. RESULTS: The clinical phenotypes of the 12 patients have mainly included cardiovascular, musculoskeletal, craniofacial, skin, ocular and other systemic signs. Four patients (patients 5-1, 5-2, 6, 7) have carried heterozygous missense variants of the TGFBR1 gene, 5 patients (patients 1-1, 1-2, 2, 3, 4) have carried heterozygous variants of the TGFBR2 gene, and 2 patients (patients 8-1, 8-2) had carried heterozygous frameshift variants of the TGFB3 gene. One patient (patient 9) had carried a heterozygous missense variant of the SMAD3 gene. Among these, TGFBR1 c.603T>G (p.1201M) and TGFB3 c.536delA (p.H179FS35) had not been reported previously. CONCLUSION Variants of the TGFBR1, TGFBR2, SMAD3, TGFB2, TGFB3 and SMAD2 genes are mainly associated with LDS. The severity of the disease phenotype caused by the same variant may vary, whilst the clinical phenotype caused by different variant sites may be specific.
Humans ; Loeys-Dietz Syndrome/genetics* ; Receptor, Transforming Growth Factor-beta Type I/genetics* ; Receptor, Transforming Growth Factor-beta Type II/genetics* ; Transforming Growth Factor beta3 ; Face

Objective:To investigate the differences of umbilical vein diameter(D), time average peak velocity(TAmax) and blood flow between congenital heart disease and normal fetus.Methods:The umbilical vein diameter and time average peak velocity of 69 fetuses with congenital heart disease (disease group) from 22 to 27 weeks were prospectively studied in Maternal-Fetal Medical Center in Fetal Heart Disease of Beijing Anzhen Hospital from May 2021 to September 2021. Q 1 (umbilical venous blood flow) was calculated according to the formular [Q=0.5TAmax·π·(D/2) 2)], and Q 2 (Q 2=Q 1/weight) was calculated according to the fetal weight. At the same time, 111 normal fetuses with matched gestational age were selected as control group. The differences of fetal umbilical vein D, TAmax, Q 1 and Q 2 between the two groups were analyzed. Results:The inner diameter of umbilical vein D, TAmax, Q 1 and Q 2 in the congenital heart disease group were lower than those in the control group(all P<0.05). In the control group, the inner diameter of umbilical vein D, TAmax and Q 1 increased with the increase of gestational age and showed a positive linear correlation( r=0.608, 0.320, 0.626; all P≤0.001), while there was no obvious linear correlation between Q 2 and gestational age( r=0.189, P=0.047). Conclusions:The decrease of umbilical vein D, TAmax, Q 1 and Q 2 in the fetus with congenital heart disease indicates the decrease of effective blood flow in placenta-fetus circulation, which indirectly reflects the decrease of placental function in the fetus with congenital heart disease.

Objective:To summarize the etiological mechanism, echocardiographic and clinical features of fetal cardiomyopathies (FCMs).Methods:According to the data of echocardiography in Maternal-Fetal Medicine Center in Fetal Heart Disease of Beijing Anzhen Hospital during 2015 January to 2020 December, 70 cases with FCMs were retrospectively reviewed, and the clinical, ultrasonic, pathological and clinical outcome data were collected. Whole exome sequencing and whole genome sequencing were used to identify the genetic changes.Results:Primary FCMs were diagnosed in 55 cases (78.6%, 55/70), including 39 fetuses with non-compaction of the ventricular myocardium (NVM), 10 with dilated cardiomyopathy (DCM), 5 with hypertrophic cardiomyopathy (HCM), and 1 with restricted cardiomyopathy (RCM). Secondary FCMs were diagnosed in 15 cases (21.4%, 15/70), including 7 fetuses with maternal anti-Ro/La antibodies (presenting with DCM), 4 with twin-twin transfusion syndrome (2 with DCM and 2 with HCM), 2 with fetal anemia (presenting with DCM), 1 with maternal diabetes (presenting with HCM) and 1 with chorioangioma of the placenta (presenting with DCM). In all cases, 9 cases were born, 3 cases died in perinatal period, and 58 pregnancies were terminated due to ineffective treatment or the decisions of pregnant women. Thirty cases with primary FCMs were performed with genetic tests, and 13 of them were identified with positive genetic changes related to FCMs, including 12 cases with NVM and 1 with HCM.Conclusions:Primary FCMs are more common than secondary FCMs in fetal period. The genetic disorders have a high proportion in fetal NVM. Fetal DCM and HCM have a large spectrum of intrinsic and extrinsic causes.

Objective:To analyze the clinical characteristics and treatment of tricuspid valve prolapse caused by chordal rupture complicated with persistent pulmonary hypertension in neonates.Methods:The clinical data of a male neonate with tricuspid valve prolapse complicated with persistent pulmonary hypertension admitted to the Neonatal Intensive Care Unit of Children′s Hospital of Hebei Province in November 2018 was analyzed retrospectively. Related literature up to September 2020 was searched with the strategy of "(neonate OR newborn) AND (tricuspid valve prolapse) AND (rupture OR necrosis) AND (papillary muscle OR chordae tendineae) AND (pulmonary hypertension)" in Wanfang, CNKI and PubMed database in Chinese and English. The characteristics of the disease were summarized.Results:A male full-term neonate was admitted due to presenting severe cyanosis for 9 hours. He was born by caesarean section and presented severe cyanosis and dyspnea at 10 min of ages, unresponsive to the positive airway pressure resuscitation. After 9 hours of mechanical ventilation, there was no improvement. Thus he was transferred to Children′s Hospital of Hebei Province. On admission, the initial blood gas analysis showed an arterial partial pressure of oxygen of 22.5 mmHg (1 mmHg=0.133 kPa). The echocardiography revealed prolapsed anterior leaflet of tricuspid valve, severe tricuspid regurgitation (TR) and pulmonary artery hypertension, and right to left shunt via a patent foramen ovale. The arterial duct was closed. The chest X-ray was normal. The boy was treated with nitric oxide, milrinone, and continued mechanical ventilation initially. Addition of prostacyclin analog (treprostinil) on day 3 led to significant improvement of pulmonary blood flow, oxygenation, and stabilization, so that the extracorporeal membrane oxygenation therapy was avoided. At 11 months after birth, the boy underwent cardiac surgery. At surgery, the rupture of chordal tendineae in anterior leaflet of tricuspid valve was found. Tricuspid annuloplasty, valvuloplasty and repair of patent foramen ovale were successfully performed. The follow-up echocardiogram at postoperative 3 months showed only mild tricuspid insufficiency. The boy was well at last follow-up at 22 months of age with normal cognitive skill development. According to literature, 20 cases of papillary muscle or chordae tendineae rupture in neonates had been reported in 12 English papers. Among the total 21 neonates, there were 12 male infants and only one premature infant with gestational age of 33 weeks. They presented with profound cyanosis soon after birth. All of them received endotracheal intubation and mechanical ventilation. Other treatments included inhalation of nitric oxide, intravenous milrinone, vasoactive drugs, diuretics and prostacyclin, etc. Extracorporeal membrane oxygenation (ECMO) was used in 6 infants as a bridge to surgical treatment. Two cases reported earlier death of cardiopulmonary failure without operation and the rest 19 survived after surgery. The followed surgery or autopsy revealed that all of them had tricuspid valve prolapse, rupture of papillary muscle or chordae tendineae.Conclusions:The severe TR resulting from rupture of papillary muscle or chordate tendineae in neonates is rare and could cause severe hypoxemia. Early recognition, adequate cardiopulmonary support to stabilize the hemodynamic status and timely surgery can significantly reduce the mortality.

Objective To establish the reference intervals (RIs) of the four biochemical parameters for liver function [aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), and alkaline phosphatase (ALP)] in adults by using the indirect method based on the data of subjects undergoing physical examination in laboratory information system (LIS). Methods With the help of the LIS, related data were collected from the healthy adults who underwent physical examination in Physical Examination Center of The First Hospital of Jilin University from October 2019 to October 2020. The Kolmogorov-Smirnov test was used to check the normal distribution of the original data; data with skewed distribution were transformed into data with approximate normal distribution using the BOX-COX method; the Turkey method was used to remove outliers; the Mann-Whitney U test or the Z -test was used for comparison between groups; the non-parametric method was used to calculate RIs. Results A total of 27 218 subjects were included after the removal of outliers. All four parameters showed age and sex differences. The RIs of AST were 14-39 U/L for male individuals aged 20-79 years, 12-32 U/L for female individuals aged 20-49 years, and 14-39 U/L for female individuals aged 50-79 years; the RIs of ALT were 10-71 U/L for male individuals aged 20-54 years, 10-49 U/L for male individuals aged 55-79 years, 7-43 U/L for female individuals aged 20-49 years, and 9-49 U/L for female individuals aged 50-79 years; the RIs of GGT were 11-70 U/L for male individuals aged 20-64 years, 10-64 U/L for male individuals aged 65-79 years, 6-45 U/L for female individuals aged 20-49 years, and 7-54 U/L for female individuals aged 50-79 years; the RIs of ALP were 38-96 U/L for male individuals aged 20-79 years, 33-89 U/L for female individuals aged 20-49 years, and 40-106 U/L for female individuals aged 50-79 years. The RIs of AST, ALT, and GGT established in this study were similar to those in the industry standards of China (relative deviation < reference change value), and the RIs of all four biochemical parameters were verified by applicability. Conclusion The RIs of the four biochemical parameters for liver function established by the indirect method are relatively consistent with those established by the direct method, which holds promise for application in clinical laboratory.

Objective:To observe the clinical efficacy of levosimendan in the treatment of patients with acute myocardial infarction (AMI) combined with cardiogenic shock (CS) and its effects on monitoring hemodynamic parameters of pulse index continuous cardiac output (PICCO).Methods:One hundred and six patients with AMI combined with CS admitted and treated in Jiaxing Second Hospital from June 2017 to December 2019 were divided into the control group and observation group according to the random number table method , with 53 cases in each group. The control group received routine comprehensive intervention, while the observation group received levosimendan treatment based on the control group. In observation group, 12 μg/kg of levosimendan was administered intravenously within 10 min, then, 0.1 μg/(kg·min) was administered intravenously and continued for 24 h. In control group, 5% glucose injection was administered intravenously, and the intravenous infusion rate and time was same as that in observation group. Both groups were treated for 24 h. PICCO was used to monitor the hemodynamic parameters , including heart rate (HR), central venous pressure (CVP), cardiac index (CI), global end-diastolic volume index (GEDVI) and extravascular lung water index (EVLWI) before and after the treatment; the neurohumoral indexes including norepinephrine (NE), angiotensinⅡ(AngⅡ); cardiac function indexes including stroke volume (SV), left ventricular ejection fraction (LVEF), and the efficacy and complications were observed and compared between two groups.Results:After 24 h of treatment, the levels of HR, CVP, GEDVI, EVLWI in two groups were decreased, and the levels of above index in the observation group were lower than those in the control group: (90.26 ± 12.61) beats/min vs. (97.82 ± 12.58) beats/min, (9.85 ± 1.14) cmH 2O (1 cmH 2O = 0.098 kPa) vs. (11.63 ± 1.37) cmH 2O, (759.53 ± 62.47) ml/m 2 vs. (867.21 ± 63.24) ml/m 2, (7.95 ± 1.56) ml/kg vs. (9.01 ± 1.78) ml/kg; after treatment the level of CI in the observation group was higher than that in the control group: (3.58 ± 0.74) L/(min·m 2) vs. (2.37 ± 0.86) L/(min·m 2), and the differences were statistically significant ( P<0.05). After 24 h of treatment, the levels of NE and AngⅡ in two groups was decreased, and the levels of NE and AngⅡ in the observation were lower than those in the control group: (60.42 ± 5.93) ml vs. (54.42 ± 6.14) ml, (41.62 ± 4.19)% vs. (36.87 ± 4.36)%, and the differences were statistically significant ( P<0.05). After 24 h of treatment, the time of intra-aortic balloon counterpulsation (IABP) in the observation was shorter than that in the control group: (61.52 ± 15.41) h vs. (89.56 ± 17.63) h; the injury rate of renal functions in the observation was lower than that in the control group: 3.77%(2/53) vs. 18.87%(10/53), and the differences were statistically significant ( P<0.05). There was no significant difference in mortality and other complications after 30 d of follow-up between two groups ( P>0.05). Conclusions:Levosimengdan can significantly improve the PICCO hemodynamics, neurohumoral indexes and cardiac function indexes of patients with AMI and CS. It has a protective effect on the kidneys of patients, but it cannot significantly improve the 30-day mortality rate of patients.