OBJECTIVE:To prepare Levofloxacin and triamcinolone acetonide double-loaded ophthalmic gel. METHODS:Us-ing levofloxacin hydrochloride and triamcinolone acetonide as main components,carbopol-940P as base material,HPMC K4M as tackifier,Levofloxacin and triamcinolone acetonide double-loaded ophthalmic gel was prepared. Using dissolution time as index, the contents of carbopol-940P and HPMC K4M were determined by single factor test,and dissolution time,viscosity and the con-tents of 2 main components were also determined. RESULTS:The concentrations of carbopol-940P and HPMC K4M were 0.4%and 1.2%,separately. The dissolution time was more than 24 h and viscosity was 1 142.67 Pa·s. The content of levofloxacin hydro-chloride was 97.3% of labelled amount (RSD=0.84%,n=3),and that of triamcinolone acetonide was 92.97% of labelled amount(RSD=1.32%,n=3). CONCLUSIONS:Levofloxacin and triamcinolone acetonide double-loaded ophthalmic gel has been prepared successfully.

Objective: To investigate the BRCA1 and BRCA2 gene mutations in patients with recurrent serous ovarian cancer and their clinical significances. Methods: A total of 57 patients with recurrent serous ovarian cancer in the First Affiliated Hospital of Nanchang University from January 2016 to January 2018 were collected. High-throughput second-generation sequencing technology was used to detect BRCA1 and BRCA2 mutations in patients' blood. Statistical analysis was performed on the relationship between BRCA mutations and clinicopathological factors and prognosis. Results: Of the 57 patients with recurrent serous ovarian cancer, 16 patients (28.07%) had BRCA mutations, among which 3 patients (5.26%) had nonsense mutations, 7 patients (12.28%) had frameshift mutations, and 7 patients (12.28%) had missense mutations. There was no significant difference in BRCA mutation rate among patients with different American Obstetrics and Gynecology Union (FIGO) stage, lymphatic metastasis, tissue differentiation and age stratification (all P > 0.05). Patients with family history of ovarian cancer or breast cancer had a higher BRCA mutation rate than patients without family history (7/12 vs. 9/45, χ² = 5.13, P = 0.02), and patients who were sensitive to first-line platinum treatment had a higher BRCA mutation rate than those with drug resistant (16/45 vs. 0/12, P = 0.04). Patients with BRCA mutation had a lower serum CA125 level than patients with BRCA wild-type [(774±548)×10³ U/L vs. (1 522±1 269)×10³ U/L, t = 3.106, P = 0.003], and a longer median progression-free survival (PFS) time (20.5 months vs. 12.0 months, P = 0.01). Conclusions Serous ovarian cancer patients with BRCA mutations have higher sensitivity to platinum-based chemotherapeutic drugs and better PFS. Detection of BRCA mutations helps to judge prognosis and guide medication.

Objective To investigate the correlation between quantitative parameters with contrastenhanced ultrasound (CEUS) and microvessel density (MVD).Methods Thirty-four placenta of rat model of preeclampsia underwent CEUS examination.The peak intensity time curves on the enhanced images were analyzed quantitatively with computer to get quantitative parameters[the time to peak(TTP), maximal peak intensity(Imax), the area under curve(AUC) and the mean perfusion volume(V)].These parameters were compared with MVD counted with immunohistochemistry and the correlation was statistically studied.Results The TTP in the enhanced images was (14.55 ± 3.45)s, Imax was (20.83 ± 6.15) dB, AUC was (1868.61 ± 25.76)dB, V was (58.01 ± 23.56)dB, and the MVD of placenta of rat model of preeclampsia was (88.98 ± 24.78) in 34 rats.The Imax was correlated positively to MVD (r = 0.885, P = 0.000) ,AUC was correlated positively to MVD (r = 0.677, P = 0.001), V was correlated positively to MVD (r =0.877, P = 0.000).There was no correlation between TTP and MVD in lesions.Conclusions The Imax,AUC and V calculated with CEUS were correlated to MVD, these parameters were valuable index for quantitative evaluation of placental blood perfusion.

OBJECTIVETo detect the disease-causing mutation in a pedigree affected with autosomal dominant congenital cataract.

METHODSGenomic DNA was extracted and purified from peripheral blood samples from members of the pedigree and 100 healthy controls. Coding regions of 18 candidate genes were screened with PCR and Sanger sequencing. Identified mutations were verified among 100 healthy individuals to exclude single nucleotide polymorphisms.

RESULTSA heterozygous nonsense mutation c.471G>A of the CRYGD gene, which resulted in p.Trp157Term, was identified in all three patients. The same mutation was not found in the two normal individuals from the family and 100 healthy controls. The nonsense mutation was predicted to be "disease causing" by Mutation t@sting program.

CONCLUSIONThe nonsense mutation c.471G>A of the CRYGD gene probably underlies the congenital cataract in the pedigree.

Cataract ; etiology ; genetics ; Child ; Codon, Nonsense ; Humans ; Male ; Sequence Analysis, DNA ; gamma-Crystallins ; genetics

OBJECTIVETo identify potential mutations of PKD1 gene in a family affected with autosomal dominant polycystic kidney disease (ADPKD).

METHODSThe coding regions of the PKD1 gene were subjected to PCR and Sanger sequencing. Reverse transcription-PCR (RT-PCR) was used to determine the relative mRNA expression in the patient.

RESULTSA splicing site mutation, c.8791+1_8791+5delGTGCG (IVS23+1_+5delGTGCG), was detected in the PKD1 gene in all 5 patients from the pedigree but not in 6 phenotypically normal relatives and 40 healthy controls. Sequencing of RNA has confirmed that there were 8 bases inserted in the 3' end of exon 23 of the PKD1 gene.

CONCLUSIONThe novel c.8791+1_8791+5delGTGCG mutation has created a new splice site and led to a frameshift, which probably underlies the ADPKD in the family. Above finding has enriched the mutation spectrum of the PKD1 gene.

Adult ; Female ; Humans ; Male ; Mutation ; genetics ; Pedigree ; Polycystic Kidney, Autosomal Dominant ; genetics ; RNA Splicing ; genetics ; TRPP Cation Channels ; genetics ; Young Adult

OBJECTIVETo identify potential mutation of PHEX gene in two patients from a family affected with X-linked hypophosphatemia (XLH).

METHODSPCR and Sanger sequencing were performed on blood samples from the patients and 100 healthy controls. Reverse transcription-PCR (RT-PCR) was used to determine the mRNA expression in patient samples.

RESULTSA splicing site mutation, IVS21+2T>G, was found in the PHEX gene in both patients but not among the 100 healthy controls. RT-PCR confirmed that exon 21 of the PHEX gene was deleted.

CONCLUSIONThe novel splicing mutation IVS21+2T>G of the PHEX gene probably underlies the XLH in this pedigree. At the mRNA level, the mutation has led to removal of exon 21 and shift of the open reading frame (p.Val691fsx), resulting in premature termination of protein translation.

Adult ; Base Sequence ; DNA Mutational Analysis ; Exons ; Familial Hypophosphatemic Rickets ; genetics ; Female ; Genetic Diseases, X-Linked ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; PHEX Phosphate Regulating Neutral Endopeptidase ; genetics ; Pedigree ; RNA Splicing ; Young Adult

OBJECTIVE: To detect potential variants of MECP2 gene in three pedigrees affected with Rett syndrome (RTT). METHODS: All exons and their flanking regions of the MECP2 gene were subjected to Sanger sequencing and multiplex ligation-dependent probe amplification assay. RESULTS: The probands of pedigrees 1 and 2 have respectively carried a c.965C>G and a c.1157_1197del41 variant of the MECP2 gene, while the proband of pedigree 3 carried a heterozygous deletional variant in exon 4 of the MECP2 gene. CONCLUSION Variants of the MECP2 gene probably underlay the RTT in the three pedigrees. Above finding has enriched the spectrum of MECP2 gene variants, and provided a guidance for the patients upon preimplantation genetic testing and prenatal diagnosis.

Objective:To observe the clinical effects of hot-wet compression with Xiaohua ointment for acne mastitis in mass stage and its impacts on humoral immune function and inflammation.Methods:85 cases of patients with acne mastitis in mass stage treated in our hospital from January 2018 to January 2019 were selected as the research objects and randomly divided into control group (42 cases) and observation group (43 cases). The control group taken Tuoli xiaodu powder and external use of purple detumescence cream, and the observation group received hot-wet compression with Xiaohua ointment additionally. All treated for 30 days. The clinical efficacy, symptom scores, breast mass size, humoral immune indexes, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were compared, and the adverse reactions were recorded.Results:After treatment, the humoral immune indexes of the two groups had no significant change ( P>0.05), but the pain score, breast tumor size, mass score, CRP and ESR were significantly decreased than those before treatment ( P<0.05); compared with the control group, the pain score, breast tumor size and mass score in the observation group were significantly lower than those in the control group ( P<0.05). The total effective rate of the observation group was 83.7%, which was significantly higher than 59.5% of the control group ( P<0.05). There were no obvious adverse reactions in both groups. Conclusions:Hot-wet compression with Xiaohua ointment is effective and safe for patients with acne mastitis in mass stage, and could improve their inflammation.

OBJECTIVETo explore the clinical application of droplet digital PCR (ddPCR) for genetic testing and prenatal diagnosis of spinal muscular atrophy (SMA) with deletion of SMN1 gene exon 7.

METHODSA total of 138 clinical samples, including 121 peripheral blood, 13 amniotic fluid, 2 umbilical cord blood and 2 chorionic villi from 56 SMA families, were tested by both ddPCR and multiplex ligation-dependent probe amplification (MLPA). Results of the two approaches were analyzed with commercial software QuantaSoft (ddPCR) and Coffalyser (MLPA), respectively.

RESULTSAmong the 138 cases, 25 had two copies, 84 had one copy, and 29 had null copy of exon 7 of the SMN1 gene. The results of ddPCR and MLPA were completely consistent.

CONCLUSIONAs a rapid, precise and economically efficient method, ddPCR will provide a new choice for genetic testing of SMA.

Adult ; DNA Copy Number Variations ; Family Health ; Female ; Gene Dosage ; Genetic Predisposition to Disease ; genetics ; Genetic Testing ; methods ; Humans ; Male ; Multiplex Polymerase Chain Reaction ; methods ; Muscular Atrophy, Spinal ; diagnosis ; embryology ; genetics ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Sequence Deletion ; Survival of Motor Neuron 1 Protein ; genetics