Objective To explore the effect of Chinese herbal medicine formula "tonifying kidney plus strengthening vital energy" on regulating CD40, CD40L protein and mRNA expression in spleen cell in mice with SHENXU. Methods The interaction of CD40L expressed by activated T cells with CD40 on monocytes, macrophages, or B cells is essential for the production of IL-12 and other cytokines. An experimental model of mice with SHENXU by corticosterone was established to observe the effect of Chinese formula on CD40, CD40L protein and mRNA expression in spleen cells or in ConA stimulated spleen cells by immunohistochemistry and RT-PCR. Results No significant differences were found among 5 groups of mice about the CD40 and CD40L protein expression in spleen cells. Both CD40 and CD40L expression detected by immunohistochemistry decreased seriously in ConA induced cells from SHENXU mice (18.9%?3.3% vs 15.8%?2.3%) as compared to that of normal controls(29.7%?4.2% vs 23.3%?1.3%), and so did the mRNA expression on ConA stimulated cells from SHENXU mice. They were restored in different degrees when the SHENXU mice were cured by different Chinese medicine. The effect of "tonifying kidney plus strengthening vital energy" was stronger than that of "tonifying kidney" and "strengthening vital energy" alone. Conclusions "Tonifying kidney plus strengthening vital energy" enhanced the expression of CD40, CD40L mRNA and protein, and this may be one of the mechanisms of improving the status of immune inhibition resulted by Corticosterone.

Objective To explore the application value of D-dimmer(DD),hs-CRP and homocysteine(Hcy) in postoperative condition monitoring in the patients with femoral neck fracture.Methods Forty cases of femoral neck fracture treated in our hospital were selected as the observation group and 40 patients with other fractures were selected as the control group.The observation group were given the surgical treatment,while the control group adopted the corresponding measures for conducting intervention according to the fracture situation.The DD,hs-CRP and Hcy levels were detected in the two groups.Then the detection results were compared between the two groups.Results The various indexes before treatment in the observation group were slightly higher than those in the control group without statistical difference(P>0.05).The levels of various indicators at postoperative 24,48 h in the observation group were significantly elevated,moreover the increase range at postoperative 24 h in the observation group was maximal,which was significantly higher than that in the control group(P<0.05).The positive rates of hs CRP,Hcy and DD in the observation group were 75.00%,77.50% and 60.00% respectively,while which in the control group were 0.00%,2.50% and 0.00%,the observation group was significantly higher than the control group (P<0.05).Conclusion Hs CRP,Hcy and DD can be used as the important indicators of condition monitoring for femoral neck fracture.

Objective: To explore early diagnostic and resent prognostic assessment value of serum levels of ischemia-modified albumin (IMA) and cardiac troponin I (cTnI) for acute coronary syndrome (ACS).Methods: A total of 175 ACS patients were selected, including 73 cases with unstable angina pectoris (UAP), 61 cases with non-ST elevation myocardial infarction (NSTEMI) and 41 cases with ST elevation myocardial infarction (STEMI).According to chest pain-to-visit time, ACS patients were divided into <3h group (n=112) and 3~6h group (n=63);another 40 healthy subjects were selected simultaneously as healthy control group.Serum IMA and cTnI levels were compared among above groups and between patients suffering from major adverse cardiovascular events (MACE) within 30d or not in <3h group.Risk factors for MACE in <3h patients within 30d were screened.Results: Compared with healthy control group and UAP group, there were significant rise in serum levels of IMA[(16.78±4.25) μg/L, (35.16±8.32) μg/L vs.(49.76±9.29) μg/L, (52.07±11.34) μg/L], cTnI[(0.17±0.06) ng/ml, (0.15±0.06) ng/ml vs.(7.65±1.29) ng/ml, (8.83±1.40) ng/ml]in NSTEMI group and STEMI group, and IMA level in STEMI group was significantly higher than that of NSTEMI group, that of UAP group was significantly higher than that of healthy control group (P<0.05 or <0.01);serum IMA level of <3h group was significantly higher than those of 3~6h group and healthy control group, and cTnI level of <3h group was significantly lower than that of 3~6h group (P<0.01 all);serum levels of cTnI and IMA in patients suffering from MACE in <3h group were significantly higher than those of patients without MACE (P<0.01 both);multi-factor Logistic regression analysis indicated that elevated serum IMA level was an independent risk factor for MACE within 30d in ACS patients[OR=2.757,95%CI(2.084~4.705), P=0.001].Conclusion: The levels of cTnI and IMA significantly rise in ACS patients.IMA level possesses early diagnosis and recent prognosis evaluation value.

Objective To analyze the genetic changes and detailed clinical presentations of 5 maturity-onset diabetes of the young (MODY) cases in order to enhance the knowledge about MODY in children.Methods Seventy-eight patients initially diagnosed as diabetes mellitus between January 1 and December 31,2015 in Capital Institute of Pediatrics were retrospectively studied.Nine of them were suspected of MODY,and 5 patients were diagnosed as MODY through gene test.Clinical informations were collected including age,gender,main complaint,family history,body mass index (BMI),fasting blood glucose,fasting blood insulin,2-hour blood glucose and insulin after oral glucose tolerance test and glycosylated hemoglobin.The blood glucose was monitored dynamically in 2 patients.Targeted capture panel was designed to capture the 16 genes related to MODY,including 12 genes from MODY1 to MODY13 type and 4 genes with weak evidence of MODY according to Human Gene Mutation Database Exome capture,and Next-Generation sequencing on a HiSeq2000 (Illumina) was performed.After bioinformatics analysis,all prioritized variants detected in patients were validated by Sanger sequencing,including the probands and their parents.Results Five patients were confirmed as MODY by molecular diagnosis,accounting for 6.4％ of all the 78 patients in 2015.The ratio of male to female was 2 ∶ 3.The ages at diagnosis ranged from 2 to 11 years old,and the median age was 3 years old.Two cases were found to have abnormal blood glucose in physical examination.The rest 3 cases were discovered with abnormal blood glucose during hospitalization because of pneumonia (1 case)or diarrhea (2 cases).In 4 cases,their mothers had gestational diabetes history,in 1 case the father suffering from diabetes.BMI ranged 15.68-23.40 kg/m2.Fasting blood glucose was 6.3-7.2 mmol/L.Fasting blood insulin was 0.5-8.0 IU/L.Glucose tolerance test results showed that blood glucose of the patients was 8.6-10.8 mmol/L after 2 hours.The level of glycosylated hemoglobin was 5.5％-6.7％.Blood glucose was 3.9-13.0 mmol/L.All the 5 confirmed patients were caused by GCK gene mutation (MODY2 type).The mutations detected were located at Exon7 (2 cases),Exon4 (1 case),Exon5 (1 case),and Exon10 (1 case).Conclusions All the confirmed MODY patients were identified either through medical exam or infectious disease,and all had positive family history.Their BMI ranged widely.Fasting blood glucose was slightly elevated and glycosylated hemoglobin was normal or slightly elevated,but fasting blood insulin was normal in all the patients.Abnormal glucose tolerance test results were found in all 5 patients.Glycosylated hemoglobin was normal or slightly elevated.MODY2 was the only subtype detected in this group,which indicated that the common type in children was different from that in adults.

Objective Congenital nephrogenic diabetes insipidus (CNDI) is a rare disease, the aim of this article is to help better understanding of this disease. Methods The clinical features, genetic analysis and treatments of two siblings with CNDI were retrospectively analyzed, and related literatures were reviewed. Results Both brothers had polydispia, polyuria and low concentrate urine continuously, and they both had a mutation in AQP 2 conifrmmed with Sanger sequencing. This novel frame shift mutation caused arginine of 254 to histidine, and prolonged AQP 2 protein. Conclusions Gene analysis can help diagnosis of CNDI. Amiloride is useful option for treatment.

Objective To analyze the clinical characteristics and mutation of SLC26A3 gene of a patient with congenital chloride diarrhea in order to deepen the understanding of the disease.Methods The clinical data of the patient who was admitted in Affiliated Hospital of Capital Pediatric Institute in June 2014 were collected.Venous blood of the proband and his parents (2 mL for each) had been extracted for genomic DNA isolation.The 21 exons of SLC26A3 gene were amplified with polymerase chain reaction and screened for mutations by sequencing.Results The main clinical features of the patient included polyhydramnios,preterm,normal birth weight,watery diarrhea,low weight and severe electrolyte disturbances with hypochloremia,hypokalemia,hyponatremia and metabolic alkalosis.Renin angiotensin and aldosterone were high.His urine chloride concentration was low and fecal chloride concentration was high (＞ 90mmol/L).After oral salt substitution therapy with KCl and NaCl [3 mmol/(kg · d),4 mmol/(kg · d)],the electrolyte was better,alkalosis was alleviated,and growth and development were improved.The gene analysis revealed that the patient carried nt1631T ＞ A homozygous mutation on exon 15 which lead to Ile544Asn mutation in the predicted SLC26A3 transmembrane protein sequence,which was considered to be responsible for the functional abnormality of the Cl-/HCO3-protein.His parents were carriers of SLC26A3 gene and their clinical phenotype was normal.Conclusions Congenital chloride diarrhea is a rare autosomal recessive disorder and easily misdiagnosed.The patient of early postnatal diarrhea with persistent hypochloremia,hypokalemia,hyponatremia and metabolic alkalosis should be thought about this disease.Genetic analysis can help make the diagnosis.The prognosis is good if a patient has an early diagnosis and appropriate management.

Objective To report clinical characteristics and genetic results of two sisters suffered from congenital adrenal cortex hyperplasia (17-α-hydroxylase deficiency), and relevant literatures were reviewed. Methods Clinical manifestation and laboratory examination data of two sister cases of 17-α-hydroxylase deficiency enrolled in Capital Institute of Pediatrics in March 2016 were analyzed. Sanger sequencing and MLPA for CYP17A1 genes were performed and the parents' genes were also verified. Results The two patients were four years and 10 years old, both suffered from hypokalemia after infections, and hypergonadotrophin gonad hypofunction. One case was with slightly high blood pressure. Laboratory test results showed potassium fluctuation tendency in 1.9~4.0 mmol/L, 17-OHP and DHEA was decreased. Enhanced CT showed different degree of adrenal gland enlargement. Chromosome examination of the older sister is 46, XY. Both sisters demonstrated heterozygous mutation of CYP17A1 gene. The molecular genetic analysis suggested a c.985_987delTACinsAA from father and a deletion spanning exons 1-7 of the CYP17A1 gene from mother. Conclusion 17-α-hydroxylase enzyme deficiency can be diagnosed before adolescence. Clinical hypokalemia with unknown reason and high blood pressure may indicate the disease. The diagnosis can be confirmed with gene sequencing of CYP17A1.

Objective A 10-years-old girl with Schimke immuno-osseous dysplasia ( SIOD ) was reported and a literature review presented to provide clinical and genetic information of this rare disease. Methods Retrospective analysis of a case of SIOD in Capital Institute of Pediatrics was reported. The patient and her parents' DNA were extracted from blood for detecting SMARCALl gene mutation. Literatures of the disease were reviewed. Results The patient was a ten-years-old girl who admitted because of slow growth in height for 3 years. Herstaturewas123cm(T(p.Q149X)andc.1933C>T(p.R645C)compound heterozygous mutation. A novel nonsense c.445C>T(p. Q149X)was found. One reported missense mutations c. 1933C>T(p. R645C) was detected. We reviewed literatures and found that there were 4 confirmed cases in China including this one. All the 4 cases had the characteristic of short stature, special facial features, osseous dysplasia, pigmentations in body, and proteinuria. However the severity of the disease and genetic changes are not the same. Conclusion When a patient was admitted because of short stature, diagnosis of SIOD should be suspected if he or she also had special facial feature, osseous dysplasia, café-au-lait spots, and proteinuria. Gene test is a tool to help us to make a definite diagnosis of SIOD.

Objective:To analyze the clinical features and CYP17A1 gene mutation of 17α-hydroxylase/17, 20-lyase deficiency (17OHD). Methods:The clinical data, laboratory examination and genetic results of 6 children with 17OHD in the Department of Endocrinology, Children′s Hospital Affiliated to Capital Institute of Pediatrics from March 2014 to December 2019 were enrolled and analyzed retrospectively.Meanwhile, the clinical types of all congenital adrenocortical hyperplasia (CAH) patients were calculated and then the incidence of 17OHD was calculated.Results:The 6 cases were from 5 families, and the age at diagnosis was ranged from 1 year and 6 months to 15 years old, in which 2 cases were 46, XX and 4 cases were 46, XY.Their gender were all female.Three cases presented with hypertension (50.0%), 4 cases with hypokalemia (66.7%), and 1 case with labia mass (16.7%). The gonad developed into a testis in patients with 46, XY, and patients with 46, XX had ovarian hypoplasia.The laboratory tests revealed an decrease in the cortisol at 8 AM in all cases, ranging from 0.62 to 5.93 mg/L.Five cases displayed an increase in adrenocorticotropic hormone (ACTH) in the range of 84-271 ng/L, and 1 patient with normal ACTH (58 ng/L) had a peak cortisol of 1.75 mg/L after the ACTH challenge test.Elevated progesterone was detected in 6 patients with a normal 17 hydroxyprogesterone level.Further results proved low levels of testosterone and estradiol, and high levels of luteinizing hormone (LH), and follicle stimulating hormone (FSH). CT scan showed mild adrenal hyperplasia in all cases.Among 114 CAH patients during the same period, the incidence of 17OHD came second at 5.3%.The CYP17A1 gene mutation results indicated that 2 unrelated patients were homozygous mutation for p. Y329fs (c.985_987delTACinsAA), 2 siblings were compound heterozygous mutations for p. Y329fs and exon 1-7 deletion, 1 patient was compound heterozygous mutations of p. Y329fs and p. R416C (c.1246C>T), and 1 patient was homozygous mutations for p. L465P (c.1394T> C), which was first reported in China. Conclusions:17OHD is not rare in CAH.Female children with hypokalemia, hypertension, and hypogonadism can lead to diagnostic suspicion of 17OHD.The p. Y329fs mutation in Chinese 17OHD children is a hotspot.The p. L465P (c.1394T>C) mutation is a new mutation in China and it could enrich the mutant spectrum of CYP17A1 gene in China.

The ompA gene of Chlamyia psittaci in cows was amplified by PCR with primers designed based on those reported in GenBank.The amplified ompA gene was inserted into the bacterial plasmid vector pGEX-4T-1 and then transformed into E.coli BL21(DE3) with IPTG induction. The gene was derived from plasmid pMD18-T vector and then sequenced.It was demonstrated that this recombinant fusion protein of approximately 68kD in molecular mass was highly expressed in inclusion body and more pure proteins would be produced after purification.The fusion protein specifically reacted with positive sera of bovine Chlamydia as demonstrated by Western blotting. These results indicate that this recombinant fusion protein shows good reactivity and could be used to develop the diagnostic kit for bovine Chlamydia and genetic engineering vaccine.