AIM:To study the changes of tear film and ocular surface after the coaxial micro incision 2. 2mm and 2. 8mm in the phacoemulsification.
METHODS:Eighty-six patients ( One hundred and six eyes ) from 2014/06 to 2016/01 in our hospital were enrolled. The patients were randomly divided into two groups. Forty-four patients ( Fifty-three eyes) in group A: coaxial 2. 2mm micro- incision phacoemulsification cataract extraction and intraocular lens(IOL) implantation;Forty-two patients ( Fifty-three eyes ) in group B: the conventional coaxial 2. 8mm small incision phacoemulsification cataract extraction and IOL implantation. The break up time ( BUT) , dry eye symptom ( DES) score, Schirmer's I test ( SⅠt ) and lid-wiper epitheliopathy ( LWE ) score were assessed preoperatively and postoperatively.
RESULTS:At 1wk, 1 and 2mo postoperatively, the BUT in two groups decreased after operations, and the BUT of group B was significantly lower than those of group A, the differences were statistically significant ( t = 3. 098, 4.512, 4.329; all P < 0. 05 ). The DES score in two groups increased, the DES score of group B was significantly higher than those of group A and the differences were statistically significant (t=-9. 449, -10. 029, -7. 141; all P<0. 05). The SⅠt in two groups increased after operations, the SⅠt of group B was significantly higher than those of group A and the differences were statistically significant (t=-6. 293, -4. 009, -3. 283; all P<0. 05). The LWE score in two groups increased after operations, the LWE score of group B was significantly higher than those of group A and the differences were statistically significant (t=-6. 542,-5. 125, -3. 632; all P<0. 05). At 6mo postoperatively, compared with the preoperative data, the BUT, DES score, S Ⅰ t and LWE score in group A showed no statistically significant differences (t=0. 659, 1. 276, 0. 548, 0.169; P>0. 05). The BUT, DES score, SⅠt and LWE score in group B showed statistically significant differences (t=-4. 063, 7. 306, 3. 621, 4. 208;all P<0. 05).CONCLUSION:Ocular surface has less damage and tear film has little influence at early stage after the coaxial 2.2mm microincision phacoemulsification, compared with the conventional coaxial 2. 8mm incision phacoemulsification surgery.

Objective To evaluate effects of tea polyphenols on the mRNA and nucleoprotein expression of Nrf2/Bach1 in human skin fibroblasts (HSFs).Methods Some HSFs were incubated with tea polyphenols at different concentrations of 0,2.5,5,10,20 and 40 mg/L for 24 hours.Methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferative activity of HSFs to screen the optimal concentration of tea polyphenols.Then,some other HSFs were treated with tea polyphenols at this optimal concentration for 24 hours.Real-time quantitative PCR (RT-qPCR) was performed to determine mRNA expression of Nrf2 and Bach1,Western blot analysis to measure nuclear expression of Nrf2 and Bach1 proteins,and immunofluorescence assay to determine the distribution of Nrf2 and Bach1 protein in the cell nucleus.Results MTT assay showed that 5 mg/L tea polyphenols had no obvious effects on the proliferation of HSFs,so 5 mg/L was chosen as the optimal concentration of tea polyphenols for subsequent experiments.HSFs cultured without tea polyphenols served as control group.After the treatment,the 5-mg/L tea polyphenol group showed significantly decreased mRNA and nuclear protein expression of Bach 1 (mRNA:0.629 ± 0.077 vs.0.940 ± 0.033,t =6.397,P ＜ 0.05;protein:1.424 ± 0.171 vs.16.966 ± 1.702,t =15.730,P ＜ 0.05),but significantly increased mRNA and nuclear protein expression of Nrf2 (mRNA:1.467 ± 0.076 vs.0.977 ± 0.091,t =7.133,P ＜ 0.05;protein:6.929 ± 0.121 vs.3.537 ± 0.126,t =33.636,P ＜ 0.05) compared with the control group.Immunofluorescence assay showed increased accumulation of Nrf2 protein,but decreased accumulation of Bach1 protein in the nucleus.Conclusion Tea polyphenols can promote the mRNA and nuclear protein expression as well as nuclear distribution of Nrf2,but suppress the mRNA and nuclear protein expression as well as nuclear distribution of Bach 1,finally exerting antioxidative effects.

To characterize the genomic sequence and arrangement of WU polyomavirus (WU virus) identified in clinical specimens collected from children with acute respiratory infections in Beijing, China, the sequences of capsid proteins VP1, VP2, and the large tumor antigen (LTAg), as well as the 5'-terminal sequence of WU virus, were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification. The PCR amplicons were sequenced, and genomic sequence analysis was performed by using the software DNAStar. In addition, VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus. The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences (CDS) sited on one strand coding for capsid proteins VP2, VP3 and VP1, and two CDS sited on the complementary strand coding for small tumor antigen (STAg) and LTAg; These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software, indicated that these sequences coming from children in Beijing shared high homology (over 98.8%) with those from GenBank. Phylogenetic analysis of these VP2 CDS by using Neighbor-joining (NJ) analyses with 2,000 bootstraps (Mega 4.0) showed that 20 sequences out of 22 belonged to clade Ia, and other 2 of them belonged to clade III, including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc. In conclusion, the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement. Overall variation of VP2 CDS was very low, and there were different clades circulating in Beijing with a dominant clade Ia, which is different from dominated Ib circulating in other parts of the world reported previously, and a novel clade IIIc was proposed.
Acute Disease ; Child, Preschool ; China ; Female ; Genome, Viral ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomavirus ; classification ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Viral Proteins ; genetics

OBJECTIVETo determine the causative gene mutation in a Chinese family with split hand/split foot malformation (SHFM) and explore the genotype-phenotype relationship.

METHODSGenomic DNA was extracted from peripheral blood samples of the patients and their family members. Polymerase chain reaction (PCR) was performed to amplify all the exons of P63 gene and HOXD13 gene. Then the PCR products were sequenced bidirectionally to screen mutations.

RESULTSA heterozygous 956G>A transversion in exon 7 of P63 gene was identified in all patients, which resulted in the substitution of histidine residue for arginine at position 280 of P63 protein (R280H). This mutation was not found in the unaffected family members.

CONCLUSIONPatients in this pedigree are characterized by symmetrical split hand and split foot with syndactyly. This condition is caused by the R280H mutation in P63 gene.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; DNA Mutational Analysis ; Female ; Genetic Diseases, X-Linked ; genetics ; Genotype ; Homeodomain Proteins ; genetics ; Humans ; Infant, Newborn ; Limb Deformities, Congenital ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Pedigree ; Phenotype ; Syndactyly ; genetics ; Transcription Factors ; genetics

OBJECTIVETo study the therapeutic effects of different doses of botulinum toxin A (BTX-A) injection on tiptoe deformation in children with cerebral palsy.

METHODSA total of 256 children with tiptoe deformation due to spastic cerebral palsy were classified into group A (muscle tension levels I-II, n=147) and group B (muscle tension levels III-IV, n=109). Group A was randomly divided into group A1 (injected with high-dose BTX-A, n=73) and group A2 (injected with low-dose BTX-A, n=74). Group B was randomly divided into group B1 (injected with high-dose BTX-A, n=55) and group B2 ( injected with low-dose BTX-A, n=54). The dose of BTX-A was 6 U/kg for groups A1 and B1 and was 3 U/kg for groups A2 and B2. Before the injection and at 1,2,6, and 12 months after injection, the muscle tension of limbs was evaluated with the modified Ashworth Scale, and the recovery of motor function of lower limbs was assessed with the Gross Motor Function Measure (GMFM).

RESULTSBefore and after treatment, there were no significant differences in Ashworth and GMFM scores between groups A1 and A2 (P>0.05). After treatment, group B1 had a significantly reduced Ashworth score and a significantly increased GMFM score, and group B1 had a significantly lower Ashworth score and a significantly higher GMFM score compared with group B2 (P<0.05). For groups A and B, Ashworth score gradually declined post-treatment, reached the lowest point at 3 months after treatment, and returned to the level before treatment at 12 months after treatment; GMFM score gradually increased post-treatment and reached the peak level at 12 months after treatment (P<0.05).

CONCLUSIONSThe level of muscle tension should be considered when BTX-A injection is used for treating tiptoe deformation in children with cerebral palsy. It makes no difference to use high- or low-dose BTX-A when the muscle tension level is within I-II, but high-dose BTX-A has a better performance in reducing muscle tension and improving motor function when the muscle tension level is within III-IV.

Botulinum Toxins, Type A ; administration & dosage ; Cerebral Palsy ; complications ; drug therapy ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Injections ; Male ; Muscle Tonus ; drug effects ; Toes ; abnormalities

OBJECTIVETo study the clinical efficiency, electroencephalogram (EEG) changes and cognitive improvements of ketogenic diet (KD) in children with refractory epilepsy.

METHODSTwenty pediatric patients (7-61 months in age) with refractory epilepsy were recruited between August 2012 and August 2013. KD therapy was performed on all participants for at least 3 months based on a fasting initiation protocol with the lipid-to-nonlipid ratio being gradually increased to 4 : 1. Seizure frequency, type and degree were recorded before and during KD therapy. A 24 hours video-electroencephalogram (V-EEG) examination and Gesell Developmental Scale assessment were performed prior to KD therapy, and 3, 6, 9 months after KD therapy.

RESULTSSix patients became seizure free after KD therapy, with a complete control rate of 30%. Seizure frequency reduction occurred in 13 (65%) patients, EEG improvement in 8 (40%) patients, and improvement in Gesell Developmental Scales (gross motor and adaptability in particular) in 6 (30%) patients. The KD therapy-related side effects were mild.

CONCLUSIONSKD therapy is safety and effective in reducing seizure frequency and improving EEG and cognitive function in children with refractory epilepsy.

Child, Preschool ; Diet, Ketogenic ; adverse effects ; Electroencephalography ; Epilepsy ; diet therapy ; physiopathology ; Female ; Humans ; Infant ; Male ; Prospective Studies ; Recurrence

OBJECTIVETo study the clinical features of non-epileptic seizures associated with cerebral palsy (CP) in children.

METHODSA total of 1 198 children with CP (age: 9 months to 6 years) were enrolled. The children with paroxysmal events were monitored by 24 hrs video-EEG (VEEG) to make sure the seizures were epileptic or non-epileptic. The symptoms, age, CP types and EEG features were observed in children with non-epileptic CP.

RESULTSFive hundred and seventy-eight children (48.24%) presented paroxysmal events. The seizures were epileptic in 231 children (19.28%) and non-epileptic in 322 cases (26.88%). In the 322 cases of non-epileptic CP, the paroxysmal events were of various kinds, including non-epileptic seizure tonic, seizure shake head, shrug shoulder or head hypsokinesis, cry or scream, panic attacks, sleep myoclonic and stereotyped movement. One hundred and fifty-eight (49.1%) out of the 322 children demonstrated nonspecific EEG abnormalities. One hundred and eleven children (34.5%) were misdiagnosed as epilepsy in primary hospitals. The CP children less than one year old showed higher frequency of non-epileptic seizures than the age groups over 1 year and 3 to 6 years. The frequency of non-epileptic seizures was the highest in children with spastic CP (168 cases, 52.2%), followed by dyskinetic CP (69 cases, 21.4%) and mixed type CP (65 cases, 20.2%).

CONCLUSIONSThe paroxysmal events in children with CP partially are non-epileptic seizures and it is important to differentiate non-epileptic from epileptic seizures. The frequencies of non-epileptic seizures may be associated with a child's age and CP type.

Cerebral Palsy ; Diagnostic Errors ; Electroencephalography ; Epilepsy ; diagnosis ; Humans ; Seizures ; diagnosis

To understand the role of respiratory syncytial virus (RSV) in children with acute respiratory infections (ARI) in Tibet Autonomous Region and the contribution of two major groups of RSV, nasopharyngeal aspirates (NPA) were collected from hospitalized children with ARI in Department of Pediatrics, Tibet People's Hospital in Lasa, Tibet from April to July in 2011 and tested for seven common respiratory viruses and human metapneumovirus (hMPV) by direct immunofluorescence assay (DFA). Total RNAs were extracted from RSV positive samples by DFA and reverse transcripted to cDNA. Nested-PCR was employed to determine the genogroups of RSV, which were confirmed by real time-PCR and sequence analysis for G protein encoding gene. The Characteristics and variations of G genes from RSV in this project were identified by sequence comparison with those G genes in GenBank. Out of 167 samples, 65 were positive for respiratory viruses with a total positive rate of 38.9%, including 45 (69.2%, 45/65)positive samples for RSV. Among 42 samples that were positive for RSV and genotyped, 40 were identified as group A and 2 as group B. Sequence analysis of full-length G genes for 7 RSV of group A indicated that all of these belonged to subgroup GA2. The nucleotide identities between RSVs from Tibet and prototype A2 strain were 90.7%-91.8%, with 86.5%-87.2% identities of amino acid. The mutations of amino acids were mainly located in both ends of a highly conserved region in the ectodomain of the G proteins. The data indicated that RSV was the most important viral etiologic agent of ARI in spring of 2011 in Tibet and group A of RSV was predominant during the study period. High divergence existed in the ectodomain of G proteins of RSVs from Tibet.
Acute Disease ; Amino Acid Sequence ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Respiratory Syncytial Virus Infections ; virology ; Respiratory Syncytial Viruses ; chemistry ; classification ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Sequence Alignment ; Tibet ; Viral Proteins ; chemistry ; genetics

AIM:To study the influence of lithium chloride (LiCl) on the neuronal differentiation of rat bone marrow mesenchymal stem cells (MSCs), and to explore whether autophagy was involved in this process .METHODS:MSCs were isolated and cultured in vitro.The cells were divided into LiCl group and control group .MSCs were treated withβ-mercaptoethanol as an inducer for triggering the cells to differentiate into neurons .The expression of neuronal markers-neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2), and autophagic marker-microtubule-associ-ated protein 1 light chain 3 ( LC3) were measured by immunofluorescence method and Western blot .An autophagy activator rapamycin and autophagy inhibitor 3-methyladenine (3-MA) were applied to modulate the autophagy in the LiCl treated-cells.The protein expression of NSE and MAP-2 were determined by Western blot .RESULTS: After induction, the ex-pression of NSE and MAP-2 were increased .The percentage of NSE-and MAP-2-positive cells and the expression of NSE and MAP-2 in the LiCl group were greater than those in control group (P<0.05).After induction, the number of LC3-positive dots and the expression of LC3-Ⅱin LiCl group were greater than those in control group (P<0.05).The expres-sion of NSE and MAP-2 increased when the autophagy was modulated by rapamycin in LiCl treated -cells, and on the contra-ry, the expression of NSE and MAP-2 were inhibited as autophagy was modulated by 3-MA.CONCLUSION: Lithium chloride may promote the neuronal differentiation of rat bone marrow mesenchymal stem cells by modulating autophagy .

OBJECTIVETo investigate the viral etiology and clinical features of hospitalized children with acute respiratory tract infections in Tibet.

METHODNasopharyngeal aspirate samples were collected from children with acute respiratory tract infection hospitalized at the department of Pediatrics, Tibet Autonomous Region People's Hospital from April to July, 2011. The specimens of nasopharyngeal aspirate were screened for antigens of 7 common respiratory viruses by direct immunofluorescence (DIF) [respiratory syncytial virus (RSV), adenovirus (ADV), parainfluenza viruses type I-III, influenza virus A and B] and human metapneumovirus. Clinical data of the children were analyzed by statistical software SPSS16.

RESULTA total of 167 children with acute respiratory tract infections hospitalized from April to July 2011 were enrolled in this investigation. Sixty-five out of 167 specimens were positive for viral antigens. The virus positive rate for specimens was 38.9% (65/167). Two of 65 positive specimens were positive for 2 virus antigens (RSV + influenza B) and (hMPV + parainfluenza virus type III), respectively. RSV was detected in 45 cases (67.2%, 45/67) which was the most predominant, followed by parainfluenza virus type III detected in 7 cases (10.4%, 7/67), ADV in 6 cases (9.0%, 6/67), parainfluenza virus type I in 4 cases (6.0%, 4/67), influenza virus type B in 3 cases (4.5%, 3/67), and hMPV in 2 cases (3.0%, 2/67). In addition to clinical manifestations of pneumonia, such as cough and shortness of breath, only 3 virus positive cases (6.67%) presented with wheezing, but the signs of severe cyanosis, fine rales in lung were common. Most of the children in this study recovered soon, only a few younger children with underlying diseases or complications had severe illness.

CONCLUSIONVirus is an important pathogen for acute respiratory infections for hospitalized children in Tibet. RSV was the most predominant etiological agent, especially for those younger than 3 years old.

Acute Disease ; Adolescent ; Age Distribution ; Child ; Child, Hospitalized ; Child, Preschool ; Female ; Fluorescent Antibody Technique, Direct ; Humans ; Infant ; Male ; Nasopharynx ; virology ; Respiratory Syncytial Virus Infections ; epidemiology ; pathology ; virology ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; epidemiology ; pathology ; virology ; Tibet ; epidemiology ; Virus Diseases ; epidemiology ; etiology ; pathology ; virology ; Viruses ; classification ; isolation & purification