[Clinical characteristics and CBS gene analysis of 13 cases with classic homocystinuria].

Objective: To analyze the clinical features and CBS gene variants of 13 patients with classic homocystinuria, and the strategies of individual treatment and prevention were explored. Methods: The general information, clinical manifestations, laboratory tests, cranial images, CBS gene variants, diagnosis and therapeutic strategies of 13 patients with classic homocystinuria admitted to the Department of Pediatrics of Children's Hospital Affiliated to Zhengzhou University and Peking University First Hospital from November 2013 to June 2021 were analyzed retrospectively. Results: There were 13 patients diagnosed at the age of 10 days to 14 years, 6 were male and 7 were female. There were 3 patients detected by newborn screening and received treatment at the asymptomatic stage. There were 10 patients clinically diagnosed at the age of 5 to 14 years. Their symptoms appeared at age of 1 to 6 years. The major clinical manifestations were marfanoid features, lens dislocation and (or) myopia, developmental delay, osteoporosis, and cardiovascular diseases. Brain magnetic resonance imaging showed asymmetric infarcts in 4 patients and hypomyelination in 1 case. Increased blood methionine, plasma total homocysteine and urinary total homocysteine with normal urinary methylmalonic acid were found in 13 patients. The biochemical features were consistent with classic homocystinuria. Totally 18 variants were identified in CBS gene of 13 patients, 10 variants were novel and 8 were reported. only 1 patient was partially responsive to vitamin B6 treatment, while 12 cases were non-responsive. They were mainly treated with low methionine diet and betaine supplement. Three vitamin B6 non-responsive cases received liver transplantation at age of 3, 8 and 8 years, respectively. Their blood methionine and total homocysteine returned to normal within a week after liver transplantation. One patient died. Prenatal diagnosis was performed for a fetus when the mother was pregnant again. Two pathogenic CBS gene variants were identified from the amniocytes as same as the proband. Conclusions: The clinical manifestations of classic homocystinuria are complex and variable. Blood amino acid analysis, serum or urine total homocysteine assay and gene analysis are critical for its diagnosis. There were 10 novel CBS gene varients were identified expanding the CBS gene varient spectrum. Liver transplantation is an effective treatment. Prenatal diagnosis is important to prevent classic homocysteinuria.

[Clinical diagnosis and treatment of three cases with hyperornithinemia-hyperammonemia-homocitrullinuria syndrome].

Objective: To study the clinical characteristics, methods of diagnosis and treatment of hyperornithinemia-hyperammonemia- homocitrullinuria (HHH) syndrome. Method: From July 2011 to August 2016, 3 Chinese patients with HHH syndrome were enrolled in this study. The clinical course, biochemical features, brain MRI findings, and gene mutations were analyzed. Result: The three patients' age at onset of symptoms was 3 months to 7 years, and the age of diagonosis was 3 years and 10 months to 9 years and 10 months. All of them presented with intolerance to protein-rich foods from the infant period, development retardation and abnormal posture. Case 1 and 2 had moderate mental retardation. Serum ammonia 25-276 µmol/L (reference range<60 µmol/L), alanine aminotransferase (ALT) 20-139 IU/L (reference range 9-50 IU/L), ornithine 29.12-99.44 µmol/L(reference range 15-100 µmol/L), urinary orotic acid 1.49-29.75 mmol/mol Cr (reference range 0-7 mmol/mol Cr), uracil 6.09-103.97 mmol/mol Cr (reference range 0-1.5 mmol/mol Cr). The cranial MRI revealed lesions in the basal ganglia, abnormal white matter signal, progressive demyelination and cerebral atrophy. On their SLC25A15 gene, a novel homozygous missense mutation c. 416A>G (p.E139G) was identified in case 1, a known pathogenic homozygous nonsense mutation c. 535C>T was found in case 2 and 3. Liver transplantation had been performed when case 1 was 6 years old. Significant improvements were observed in dietary habit, mental and motor functions, and biochemical parameters. After the dietary intervention with the supplements of arginine, L-carnitine, case 2 was improved, spastic paraplegia of case 3 had no mitigation. Liver transplant was recommended. Conclusion: HHH syndrome has an aversion to protein-rich food, and the patients have recurrent vomiting and progressive neurological dysfunction. Clinical diagnosis of HHH syndrome is difficult and patients may present with incomplete biochemical phenotype. The genetic analysis is key for the diagnosis. Depending on their condition, individuals with HHH syndrome can be treated with a low-protein diet, drugs and liver transplantation.

[Two cases with generalized intracranial calcification due to hereditary folate malabsorption and literature review].

Objective: This study aimed to investigate the clinical, biochemical and genetic features of two Chinese children with hereditary folate malabsorption. Method: Clinical features, laboratory examinations, treatment and SLC46A1 gene of two cases were studied. Reports on hereditary folate malabsorption utill September of 2016 were searched and the clinical and genetic characteristics of reported cases were summarized. Result: The two patients presented with megaloblastic anemia from their infant period and seizures, psychomotor retardation and regression. In case1, mean corpuscular volume (MCV) was 100 fl. Serum folate was 9.96 nmol/L. Folate and 5-methylenetetrahydrofolate in cerebrospinal fluid were 0 and 0.01 separately. In case 2, MCV was 93.9 fl. Serum folate was 4.49 nmol/L. The concentration of folate and 5-methylenetetrahydrofolate in cerebrospinal fluid were both zero. On their brain CT, progressive bilateral symmetrical calcification was observed. On their SLC46A1 gene, four mutations were identified. Case 1 had one novel mutation, c. 1238T>C (L413P) and c. 194-195insG (p.Cys66LeufsX99). From Case 2, two reported mutations, c. 1A>T (M1L) and c. 194-195insG (p.Cys66LeufsX99) were identified. The administration of folinic acid (60 to 120 mg per day) was initiated after diagnosis. Clinical improvement and normalized hematologic markers were observed after treatment. Totally 37 cases were reported in reviewed English literature, including 30 cases with mutations on SLC46A1 gene (only one Chinese patient). All the cases had the onset in infancy. The ratio of boys to girls was 1 to 1.5. Main manifestations were characterized by megaloblastic anemia (77%), failure to thrive (50%), diarrhea (27%), psychomotor retardation (63.6%), epilepsy (27%), and infection of respiratory system (45.5%). The concentration of folate in both serum and cerebrospinal fluid was decreased (72.7% and 63.6% respectively). Hypoimmunoglobulinemia accounted for 27.3%. Most of mutations in HFM were distributed between p. 65 and p. 68 (c.194-c.204), mainly due to insertion- or deletion-related frame shifts or generation of stop codons. Oral and parenteral folinic acid treatment was effective. Conclusion: Hereditary folate malabsorption often presented with megaloblastic anemia, abnormalities of digestive and nervous system, and hypoimmunoglobulinemia with recurrent infections. Low level of serum and CSF folate and screening SLC46A1 gene are keys to the etiologic study of the patients. Early supplement with folinic acid is beneficial to the prognosis.

Magnetic anisotropy and spin-glass behavior in single crystalline U2PdSi3.

We present the magnetic and transport properties of single crystalline U(2)PdSi(3) measured with the magnetic field (H) (or measuring current, I) applied along two typical crystallographic directions, i.e. H ⊥ c-axis and H c-axis (or I ⊥ c-axis and I c-axis). For both directions, a spin-glass state is confirmed to form at low temperature with the same spin freezing temperature T(f) (=11.5 K), initial frequency shift δT(f) (=0.023) and activation energy E(a)/k(B) (=90.15 K) in zero dc field. Strong anisotropy in magnetic and transport behavior is found to be a significant feature of U(2)PdSi(3). The unusual ferromagnetic-like anomaly in ac susceptibility and dc magnetization curves around T(m)=71 K is observed in the case of H c-axis but not in the cases of H ⊥ c-axis. The characteristic temperature T(ir), below which evident irreversible magnetism originated from random spin freezing can be observed, shows much stronger field dependence for H ⊥ c-axis than for H c-axis. Moreover, an unusual finding is that the electrical resistivity measurements indicate the formation of magnetic Brillouin-zone boundary gaps and much larger magnetic scattering for I ⊥ c-axis, while the coherent-Kondo-effect-like behavior is obvious for I c-axis. We also emphasize that no resistivity minimum can be detected down to 2.5 K for either direction. The observed magnetic and transport behaviors are compared with those in polycrystalline U(2)PdSi(3) and other 2:1:3 intermetallic compounds.

[Qi deficiency and blood stasis and circulatory renin-angiotensin system as well as plasminogen activator inhibitor activity in rats with cardiac qi deficiency syndrome].

OBJECTIVE: To explore the relationship between cardiac Qi deficiency Syndrome and effect of circulatory renin-angiotensin system (RAS) and endogenous fibrinolytic system in rats with heart failure (HF). METHODS: Plasma RAS levels and plasminogen activator inhibitor (PAI-1) activity were measured in 2 groups of rats: the sham-operated group (Group A) and the HF with Heart Qi deficiency Syndrome model group (Group B) established by making arteriovenous fistula. Using radioimmunoassay and chromatography, plasma renin activity (PRA) and angiotensin II (Ang II) levels and PAI-I activity were determined before, immediately after and 30 days after the surgical operation respectively. RESULTS: After the operation was completed in Group B the left ventricular dysfunction revealed, left ventricular systolic pressure decreased and end diastolic pressure increased (P < 0.05), PRA and Ang II level and PAI-1 activity increased significantly, as compared with those in Group A and before modeling (P < 0.05). With cardiac function improved, the plasma PRA, Ang II level and PAI-1 activity lowered in Group B comparing with those immediately after modeling (P < 0.05). CONCLUSION: The activation of RAS is associated with the endogenous fibrinolyticim balance, and play an important role in endogenous fibrinolytic system dysfunction in HF with Heart Qi deficiency Syndrome and Qi deficiency-blood stasis, which was helpful to explain that cause of hypercoagulant state in HF patients and increase the risk of suffering from embolism-thrombotic diseases, and might be the pathogenetic basis of Heart Qi deficiency induced Qi deficiency-blood stasis.

Antifibrillatory effect of tetrahydroberberine.

Electric stimulation and drug-induced ventricular fibrillation (VF), monophasic action potentials (MAP), and triggered activity were studied before and after administration of tetrahydroberberine (THB) in rabbits, rats or guinea pigs. At doses of 5, 10, and 20 mg.kg-1, i.v. THB increased the ventricular fibrillation threshold, and the BaCl2-induced VF was also prevented or terminated by THB in rabbits. Centrogenic VF induced by icv aconitine in rats was inhibited by pretreatment with THB in a dose-dependent manner, whereas VF induced by iv ouabain in guinea pig was inhibited to a lesser degree. For MAP, the duration at 90% repolarization (MAPD90) was prolonged remarkably, whereas the MAPD20, the MAP amplitude, and the maximal velocity of phase 0 were shortened or decreased slightly. The amplitudes of early afterdepolarization produced by cesium chloride (CsCl) were attenuated, while the cumulative threshold doses of CsCl for sustained ventricular tachycardia were elevated by THB. These results indicated that THB had an potent antifibrillatory effect, which might be attributed to its blockade of potassium, calcium, and sodium currents.

Induction and purification of kievitone hydratase from Fusarium solani f. sp. phaseoli.

Fusarium solani f. sp. phaseoli is capable of detoxifying the major isoflavonoid phytoalexins produced by its host plant Phaseolus vulgaris. One of the enzymic activities involved is kievitone hydratase (KHase), a secreted glycoprotein which catalyses the conversion of kievitone to the less fungitoxic derivative, kievitone hydrate. Even under conditions of substrate induction, the enzyme is expressed at levels that are too low for satisfactory purification. Therefore, several other isoflavonoids were tested as inducers in culture. Among the phytoalexins produced by the host plant, phaseollinisoflavan was the best inducer, elevating the level of secreted enzyme eight-fold. Treatment with biochanin A, a product of chickpea, resulted in a 16-fold increase of secreted activity. The maximum rate of induction was observed 9-24 hr after addition of biochanin A, during which time several metabolites of the inducer were also present. KHase was purified from filtrates of biochanin A-induced cultures. Denaturing gel electrophoresis indicated that two species of Mr 47,000 and 49,000 copurified with the activity. N-Terminal sequence analysis indicated that the two species possessed related, or identical, polypeptide moieties. Comparison with the size of the non-denatured enzyme, previously determined to be ca 100,000, indicates that its native state is a dimer.

[Inhibitory action of intracerebroventricular injections of norepinephrine and clonidine on ventricular extrasystoles induced by stimulating hypothalamus in rabbits].

The effects of bilateral intracerebroventricular (icv) injections of norepinephrine (NE) and clonidine on hypothalamic stimulation-induced ventricular extrasystoles (VE) were investigated in alpha-chloralose and urethane anaesthetized rabbits. NE and clonidine reduced the number of HVE. The effects of NE were antagonized by pretreatment with phentolamine and yohimbine, but unaffected by pretreatment with prazosin. Pretreatment with yohimbine could also abolish the effects of clonidine. It is concluded that the effects of reduction in VE induced by NE and clonidine may be mediated by activating central alpha 2-adrenoceptors rather than alpha 1-adrenoceptors.

[Inhibitory effects of gamma-aminobutyric acid and diazepam on ventricular arrhythmias induced by hypothalamic electric stimulation].

The experiments were carried out on rabbits with ventricular arrhythmias (VA) induced by hypothalamic electric stimulation. The effects of GABA or diazepam (Dz) and picrotoxin on VA were observed. After GABA 0.25, 0.5, 1 mg/rabbit were injected into cerebral ventricle (icv) or cisterna magna (icm), the VA was dose-dependently reduced, the similar effect was seen after Dz 0.1 mg was injected into subarachnoid cavity (sac) or icm. The effect was not induced by GABA sac or Dz icv. Picrotoxin 20 micrograms icv or 30 micrograms icm obviously increased VA, and this effect was partly antagonized by pretreatment with GABA 0.5 mg icv or icm and Dz 0.1 mg icm. The results suggest that central GABAergic system plays an important role in development of arrhythmias, and that increasing activity in this system may inhibit VA.