OBJECTIVE To investigate the relatedness between chronic hepatitis C patients in southern Anhui Province, and the serum ALT levels, the quantity of HCV RNA and the HCV genotypes. METHODS The rate method and fluorescence quantitative PCR method were used to detect serum levels of ALT and HCV RNA in 141 cases of chronic hepatitis C patients in southern Anhui Province. Reverse transcript PCR was used to genotype in 50 cases of specimens. RESULTS Spearman rank correlation analysis was used to analyze correlation between serum ALT levels and the quantity of HCV RNA, rs=0.213, P=0.011

Objective Effects of aqueous extract from Lishi No.5 formula on concentration of intracellular free-calcium fluorescent intensity in neurons. Methods Hippocampus neurons of 1-day newborn SD rats were cultured with conventional culture technique. The cells cultured for 9-12 days were used for experiment. Intracellular free calcium fluorescent intensity of neurons cultured under different conditions was assayed with confocal microscopic calcium image technique after loading of Fluo-3/AM. Results Free-calcium concentration was enhanced by aqueous extract from Lishi No.5 formula, this concentration was up to 126.35?9.35nmol/L, but the concentration is at normal scope; L-type calcium channel blocker Nifedipine may block the effect of aqueous extract from Lishi No.5 formula enhancing intracellular free-calcium concentration in part, it make intracellular free-calcium concentration down to 90.75?10.15nmol/L, but Nifedipine itself also decreased markedly free-calcium concentration to 40.65?5.65nmol/L. NMDA increase Markedly calcium fluorescent intensity, the effects of NMDA was decreased notably after pre-treating with aqueous extract from Lishi No.5 formula. The effect of MK-801, an antagonist of NMDA receptor, on inhibition of NMDA increasing free calcium fluorescent intensity was significantly reduced after pretreatment of aqueous extract from Lishi No.5 formula. Conclusion Aqueous extract from Lishi No.5 formula maybe bidirectional adjust intracellular free calcium concentration via enhancing L-type calcium channel activity and blocking NMDA receptor in part.

Objective To study the antiviral effects of Pudilan xiaoyan oral liquid on Hep-2 cell models infected with respiratory syncytial virus (RSV), adenoviruses serotype 3 strains (ADV3) in vitro. Methods The cell cytotoxic and inhibition effect of Pudilan xiaoyan oral liquid on RSV or ADV3 were investigated by MTT assay and the inhibitory assay for cytopathic effect (CPE) in Hep-2 cell cultures to detect its antiviral effects. Results The median toxic concentration (TC50) of Pudilan xiaoyan oral liquid on Hep-2 cells was 776.97 mg/L. The median effective concentration (EC50) of inhibiting RSV and ADV3 were 28.08, 28.10 mg/L,whose therapeutic index (TI) were 27.67 and 27.65 respectively. The safety coefficient of Pudilan xiaoyan oral liquid is higher than that of control, ribavirin. Compared with the virus control group, Pudilan xiaoyan oral liquid can obviously produce actions of a dose-dependent relationship to inhibit CPE in Hep-2 cells infected with RSV or ADV3 virus. Conclusions Pudilan xiaoyan oral liquid significantly improves the protection against RSV and ADV3 virus infection in Hep-2 cells. And the inhibition of CPE induced by viruses infection increased with the elevation of higher drug concentration for its antiviral effect augmented in vitro.

Objective To assess the efficiency and reliability of clinical genetic diagnosis of methylmalonic acidemia(MMA) using new generation sequencing platform (HiSeq2000).Methods 1.Nine patients diagnosed with clinical signs of MMA were recruited.DNA library from the patients were mixed with designed gene capture probe.The whole exons region of 48 genes related to organic acid metabolism were screened using the gene capture combined with high-throughput sequencing.2.The joints were removed and the low quality data were filtered,the data were analyzed by means of SNP and InDel.To avoid the false positive,the abnormal sites were verified using the Sanger sequencing method.3.The detection of the organic acid in the urine was performed through gas chromatography-mass spectrometry and other auxiliary examinations.Results 1.Gene mutation:7 gene mutations of MMACHC were identified in 7 patients.Seven mutations:c.482G ＞ A,c.567_568insT,c.609G ＞ A,c.440_441del,c.80A ＞ G,c.315C ＞ G,c.90G ＞ Awere screened.The mutation c.440_441del had not been reported before,and others were all related to the disease.Two gene mutations of mutase apoenzyme(MUT) were identified in 1 case,all of which were introns:.c.754-1G ＞ C,c.1677-1G ＞ A.The novel mutation was c.754-1G ＞ C.No gene mutation was identified in 1 patient.2.Clinical manifestation:all of the patients were development delay,but the degrees were different;3 patients with convulsion; 1 patient with headache and central facial paralysis;1 patient with repeated intractable metabolic acidosis;1 patient with repeated hemolysis.Electroencephalogram of the all patients were abnormal;the result of cranial MRI of the 8 patients were abnormal;In all patients,urine level of methylmalonic acid significantly increased (273.4-146 022.8 times).Blood homo cysteine of 8 patients were significantly increased(27.13-396.84 μmol/L,normal ＜ 20 μmol/L).3.Sanger sequencing:there were no false positive exists.Conclusions 1.There were not a correlation between the clinical manifestation and gene mutation of the patients with MMA.The c.609G ＞ A was the hotspot mutation of MMACHC gene in Chinese patients with MMA and homocysteinemia.2.The mutations c.440_441del and c.754-1G ＞ C were presumed to be novel mutations.3.Gene capture technology combined with next-generation sequencing technology could be used to interrogate the wealth of data available in the human genome and lay the foundations for counseling of gene.This platform can be readily and timely adopted by clinical molecular diagnosis of MMA and represents a high throughput,high sensitivity,high efficiency and other characteristics approach for screening common genetic diseases.

ObjectiveTo explore the related factors influencing effect of community-based rehabilitation (CBR) on activities of daily living (ADL) in stroke patients in.Methods202 stroke patients were randomly divided into the CBR group (n=103) and control group (n=99). The patients of the CBR group treated with rehabilitation and following-up including risk factors controlled with drug, rehabilitation, health education and mental treatment, those of the control group only treated by following-up. Before and five months after treatment, patients of two groups were assessed with Barthel Index (BI), clinical never function limitation score, cognitive item of Functional Comprehensive Assessment (FCA). Multielement regression analysis was applied, the ADL score of last assessment was taken as dependent variable, group information, location, smoking, sex, age, course of diseases, education, drinking, sleeping, the first scores of FCA and BI were taken as independent variable.ResultsThe improvement of ADL of stroke patients was related with group information, drinking, course of diseases, the first scores of FCA, BI, and etc.ConclusionEarly CBR can significantly improve the ADL of the stroke patients. Cognitive deficit also has influence on ADL.

Objective: To investigate the effect of cobalt chloride (CoCl2) on transgene expression and viral particle titers in tumor cells infected by conditionally replicating adenovirus expression vector with hypoxia response elements(HRE)-regulated E1AE1B expression (Ad-5HRE-E1AE1B-RFP) and non-HRE regulated replication-deficient adenovirus expression vector (Ad-EGFP, Ad-Luc) in vitro and in vivo. Methods: Ad-5HRE-E1AE1B-RFP had five duplicated HRE and mini CMV acted as a promoter to drive E1AE1B expression and constitutive expression of RFP as reporter. The hypoxia model was optimized by exposing tumor cells to different concentrations of CoCl2. The hypoxia-inducible factor 1 alpha (HIF-1α) protein expression was determined by Western blotting. Under the optimized hypoxia model, the positive expression of exogenous gene and virus replication of Ad-5HRE-E1AE1B-RFP or Ad-EGFP-infected tumor cells were examined by conversed microscopic observation, FACS analysis and plaques formation test. Furthermore, transgene expression induced by combined application of hypoxia-inducible replicative adenovirus and replication deficient adenovirus (Ad-Luc) was also evaluated by examining the lucifererse activity in xenografted tumor models in nude mice by micro PET. Results: Western blotting results showed that CoCl2 at 0.4 and 0.08 μg/mL could stabilize and acumulate HIF-1α protein in gastric cancer SGC7901 cells, which could better mimic hypoxia condition. The microscopic observation and FACS analysis showed that CoCl2 at 0.4 μg/mL could remarkably increase the transduction efficacy of Ad-5HRE-E1AE1B-RFP, which was verified by significant increase in the percentage of positive expression of exogenous gene RFP and fluorescence intensity. But plaques formation test showed that Ad-5HRE-E1AE1B-RFP had no replication. CoCl2 0.4 μg/mL augmented the tranduction efficacy and expression levels of non-HRE regulated replication deficient adenovirus Ad-EGFP and Ad-Luc. Combined intratumoral injection of Ad-5HRE-E1AE1B-RFP and Ad-Luc significantly increased the expression of Ad-Luc in nude mice.Conclusion: CoCl2 markedly enhances transgene expression of recombinant adenovirus. However, the underlying mechanism is not only related to the CoCl2-induced hypoxia, but also probably related to regulation of gene transcription.

Objective To explore the value of cMRI and MRS in diagnosis and differential diagnosis of single brain metastases and localized high grade gliomas.Methods The cMRI (T1WI,T2WI and contrast-enhanced MRI)and MRS data of 23 cases with single brain metastases and in 28 cases with high grade gliomas confirmed by pathology were collected and analyzed retrospectively.Results cMRI:①There were no significant differences of T1WI,T2WI and constrast-enhanced MRI between single brain metastases and high-grade gliomas.②Enhanced images of single brain metastases and localized high-grade gliomas showed that there was a significant difference in the peritumoral edema area:the nodular or ring enhancement were found in single brain metastases,the outer outline of ring enhancement was smooth,and there was no enhancement at the peripheral edema area;Irregular patchy or ring enhancement were found in localized high-grade glioma tumor,both of the outer and inner edges of the ring enhancement rough,a little patchy enhancement at the peripheral edema area were found in 1 6 patients,and there was no enhancement of the edema area in the other 1 2 patients.MRS:①There were no significant differences of the tumor parenchyma in the peaks of NAA,Cr,Cho and central Lac between single brain metastases and localized high grade gliomas.②A significant difference of the peripheral edema area were observed between them.The peaks of NAA,Cr,Cho at the edema area in all single brain metastases patients were normal.But in all high-grade glioma patients,the NAA and Cr peaks were decreased,while the Cho peaks were increased.Conclusion The morphological manifestations of single brain metastases are similar to localized high grade gliomas.An obvious difference of cMRI and MRS lines exists in the peritumoral edema area between them,which could be used for differential diagnosis.

Objective To observe and analyze the effects of rehabilitation training com-bined with drugs in prevention from secondary osteoporosis patients with hemiplegia caused by stroke.Methods 120 patients with hemiplegia caused by stroke were randomly divided into obser-vation group and control group with 60 patients in each group.The patients in the control group were treated with rehabilitation training on the basis of the conventional therapy,while the pa-tients in the observation group were treated with the combined anti-osteoporosis drugs including salmon calcitonin and calcium carbonate vitamin D3 tablets based on the therapy of the control group.The bone density of femur proximal Ward River,distal radius,first lumbar spine at hemi-plegia side of research time,the first month and the third month after the treatment in the two groups were detected and compared.Results There were no significant differences in bone density of the three tested sites of patients at the research time between the two groups (P >0.05),while at the first month and the third month after the treatment,the bone density of the patients in the observation group at the three sites were significantly higher than those in the control group (P <0.05).Conclusions The combined application of drugs and rehabilitation training to the patients with post-stroke hemiplegia can gain significant effect in prevention from the osteoporosis.

Objective:To explore the early neurodevelopmental features of young children with SYNGAP1 variants and their genotype-phenotype correlation. Methods:Young children with neurodevelopmental disorders (NDDs) (< 5 years old) who were referred to the Children′s Hospital Affiliated to the Capital Institute of Pediatrics between January 2019 and July 2022 were selected as the study subjects. All children had undergone whole-exome sequencing, comprehensive pediatric neuropsychological assessment, familial segregation analysis, and pathogenicity classification. Meanwhile, young Chinese NDD children (< 5 years old) with pathogenic/likely pathogenic SYNGAP1 variants were retrieved from the literature, with information including detailed clinical and genetic testing, neurodevelopmental quotient (DQ) of the Children Neuropsychological and Behavior Scale-Revision 2016 (CNBS-R2016). Children who did not have a detailed DQ but had their developmental status assessed by a medical professional were also included. The correlation between neurodevelopmental severity, comorbidity and SYNGAP1 variants were summarized. Results:Four young NDD children carrying SYNGAP1 variants were recruited (1 male and 3 females, with a mean age of 34.0 ± 18.2 months), among whom one harboring a novel variant (c.437C>G, p. S146*). Combined with 19 similar cases retrieved from the literature, 23 Chinese NDD young children were included in our study (8 males and 10 females, 5 with unknown sex, with a mean age of 37.1 ± 14.2 months). A loss of function (LOF) variant was found in 19 (82.6%) children. All of the children had presented global developmental delay (GDD) before the age of two. In addition, 16 (69.6%) had seizure/epilepsy at the age of 27.0 ± 12.1 months, among whom 15 had occurred independent of the global developmental delay. Myoclonic and absence were common types of seizures. Compared with those with variants of exons 8 to 15, the severity of developmental delay was milder among children with variants in exons 1 to 5. Conclusion:The early neurodevelopment features of the SYNGAP1 variants for young children (< 5 years old) have included global developmental delay and seizure/epilepsy. All of the children may present GDD before the age of two. The severity of developmental delay may be related to the type and location of the SYNGAP1 variants.

Objective To observe and analyze the effects of rehabilitation training com-bined with drugs in prevention from secondary osteoporosis patients with hemiplegia caused by stroke.Methods 120 patients with hemiplegia caused by stroke were randomly divided into obser-vation group and control group with 60 patients in each group.The patients in the control group were treated with rehabilitation training on the basis of the conventional therapy,while the pa-tients in the observation group were treated with the combined anti-osteoporosis drugs including salmon calcitonin and calcium carbonate vitamin D3 tablets based on the therapy of the control group.The bone density of femur proximal Ward River,distal radius,first lumbar spine at hemi-plegia side of research time,the first month and the third month after the treatment in the two groups were detected and compared.Results There were no significant differences in bone density of the three tested sites of patients at the research time between the two groups (P >0.05),while at the first month and the third month after the treatment,the bone density of the patients in the observation group at the three sites were significantly higher than those in the control group (P <0.05).Conclusions The combined application of drugs and rehabilitation training to the patients with post-stroke hemiplegia can gain significant effect in prevention from the osteoporosis.