As one of the first resident standardized training bases, department of ophthalmology of Shanghai Ruijin Hospital participated in this reform process from 2010. Relevant rules and regulations (training management system , training scheme implementation system and evaluation system ) were strictly obeyed. When new problems emerged, under the guidance of department in charge, a series of regimens were formulated and improved gradually by Ophthalmology Professional Committee of Shanghai Resident Standardized Training Department. Based on reviewing and summarizing the work in the last 3 years, some thoughts and suggestions on the resident standardized training in future were put forward ,in-cluding how to better solve the“heavily used, lightly cultured” problem, the“disregarding medical ethics establishment”problem, the“disregarding assessment of teachers”problem and the“disregard-ing obtaining employment”problem.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

AIM: To investigate the effect of a Chinese medicime, Dan-Shao-Hua-Xian capsule, on liver fibrosis induced by CCl_4 by observation of apoptosis and cell cycle variation in the liver cells. METHODS: Animal models were produced through eight-week treatment of the rats with CCl_4, alcohol and diet of high fat/low proteins, and then administration of Dan-Shao-Hua-Xian to the rats (1 g/kg) via stomach-tube-pouring for eight weeks was performed. Liver index, serum hyaluronic acid (HA) and glutamate pyruvate transaminase (ALT) were measured and hydroxyproline (Hyp) content in urine were determined. The extent of the liver fibrosis was observed under light microscope and apoptosis and cell cycle were also examined by cytometry between the two groups. RESULTS: Compared to the liver fibrosis group, the liver index, serum HA, ALT in the treatment group decreased, the development of liver fibrosis delayed, the urine Hyp and the number of apoptosed cells and the ratio of G_0/G_1 cells increased, as well as the S phase cells decreased, yet unable to return to normal. All those changes detected were statistically significant. CONCLUSIONS: The Dan-Shao-Hua-Xian is effective in treating the CCl_4-induced liver fibrosis in rats partly by virtue of inhibition of the growth of hepatic stellate cells and induction of apoptosis.

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.

To study the effect of pulchinenoside (PULC) on the Frizzled (FZD) expression of adjuvant arthritis ( AA) rats. AA rats were prepared through the toe injection with complete Freund's adjuvant to culture fibroblast-like synoviocytes (FLS). The effect of the oral administration with PULC on the FZD8 expression was detected by the real time qPCR. The effect of FZD8 knockout on the expressions of IL-1, IL-6, IL-8 were detected by MTT and ELISA. The role of miR-375 in the abnomal expression of FZD8 was detected by the real time qPCR. The results showed signfiicant decrease in the FZD8 expression among AA rats, FLS proliferation ater FZD8 knockout and IL-1, IL-6, IL-8 expressions and notable increase in miR-375 expression after the oral administration with PULC. The up-regulated miR-375 expression can inhibit the FZD8 expression. PULC may inhibit the FZD8 expression by up-regulating the miR-375 expression.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism ; Saponins ; administration & dosage

The present study was designed to observe the expressions of bone morphogenetic protein-7 (BMP-7) and inhibitory Smads in kidney of rats with diabetic nephropathy (DN), and explore the possible mechanism of DN. Male Wistar rats weighing 180-220 g were single injected with streptozocin (STZ, 55 mg/kg body weight) for 2, 4, 8 and 16 weeks to induce DN. Blood glucose, kidney weight/body weight and 24-hour urine protein in the control and DN rats were examined; the expressions of BMP-7, Smad6 and Smad7 were detected by using immunohistochemical techniques, Western blot and real-time PCR. The results showed that blood glucose and 24-hour urine protein in DN rats were higher than that in the control rats and kidney weight/body weight was also elevated in DN rats, especially in 16-week STZ-induced rats. The expressions of BMP-7 and Smad6 proteins in DN rats were elevated, while BMP-7 mRNA expression was increased 2 weeks after STZ injection and decreased 16 weeks after STZ injection. The expressions of Smad7 protein and mRNA were elevated in DN rats 2 weeks after STZ injection and decreased 16 weeks after STZ injection. In addition, the expressions of transforming growth factor-beta1 (TGF-beta1) and collagen type I (COL-I) mRNA were increased in DN rats. These results suggest in the early stage of DN, increase in BMP-7 and inhibitory Smad expression may play a role in the feedback regulation and restrain the development of DN.
Animals ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Collagen Type I ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; Diabetic Nephropathies ; metabolism ; Kidney ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Smad6 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo explore the application value of morphology assessment of sperm from fresh semen in routine in vitro fertilization (IVF).

METHODSWe analyzed the morphology of the sperm from fresh or optimized semen samples and, based on the sperm morphology of the raw semen, allocated 908 IVF cycles due to the pure tubal factor to different groups: morphologically normal sperm (MNS) ≤ 4%, > 4% - ≤ 15%, and > 15% in Trial 1 and MNS ≤ 1%, > 1% - ≤ 2%, > 2% - ≤ 3%, and > 3%-- ≤ 4% in Trial 2. We compared the rates of fertilization, cleavage, high-quality embryo, -blastocyst formation, and pregnancy among different groups.

RESULTSThe total fertilization rate was significantly lower in the MNS ≤ 4% than in the MNS > 4% - ≤ 15% and >15% groups (74.40% vs 78.61% and 80.03%, P < 0.01). Compared with the MNS ≤ 1%, > 1% - ≤ 2%, and > 2% - ≤ 3% groups, the MNS > 3% - ≤ 4% group showed remarkably increased rates of 2PN normal fertilization (77.23%, 78.97% and 78.99% vs 85.47%, P < 0.01), cleavage (95.71%, 96.01% and 97.27% vs 98.73%, P < 0.05), and blastocyst formation (53.85%, 49.01% and 49.55% vs 63.41%, P < 0.01). No statistically significant differences were observed in the rates of clinical pregnancy, implantation, early abortion, live birth, or malformation at birth among different groups (P > 0.05).

CONCLUSIONMNS ≤ 4% affected the total rate of fertilization while MNS ≤ 3% reduced the rate of normal fertilization in IVF. However, even MNS ≤ 1% did not result in fertilization disorder or failure. Therefore, teratozoospermia alone was not an indicator of ICSI and sperm mor- phology assessment had no obvious value for predicting the rates of embryo quality, clinical pregnancy, and live birth in IVF.

Objective To explore the correlation between serum hepatitis B virus (HBV) X antigen/antibody (HBxAg-wild/HBxAb-wild,and HBxAg-mutant/HBxAb-mutant) and the disease progression in patients with chronic HBV infection.Methods A direct enzyme immunosorbent asssay (ELISA) was performed to detect HBxAb using recombinant antigen,and a double antibody sandwich ELISA assay to detect HBxAg using monoclonal antibody and specific rabbit polyclonal antibody.HBxAg-wild/HBxAb-wild and HBxAg-mutant/HBxAb-mutant were tested in sera from cases at different stages of chronic HBV infection.A chi-square test was employed to examine statistical significance.Results The positive rates of HBxAg-wild and HBxAg-mutant in the chronic asymptomatic HBV carriers,chronic hepatitis,hepatitis B-related cirrhosis and liver cancer were 6.2％ (2/32),10.7％ (3/28),28.6％ (6/21),43.6％ (17/39) and 3.1％ (1/32),10.7％ (3/28),33.3％ (7/21),48.7％ (19/39),respectively.The positive rates of HBxAb-wild and HBxAb-mutant in the above mentioned groups were 6.2％ (2/32),21.4％ (6/28),38.1％ (8/21),53.8％ (21/39)and 6.2％ (2/32),25.0％ (7/28),42.9％ (9/21),61.5％ (24/39) respectively.The positive rates of HBxAg-wild and HBxAg-mutant were not significantly different among the above groups (x2 =0.871,0.780,0.565 and 0.317,respectively; all P＞0.05) ; The positive rates of HBxAb-wild and HBxAb-mutant were also similar among all the groups (x2 =0.780,0.709,0.580 and 0.210,respectively; all P＞0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in patients with low viral loads (HBV DNA＜1 × 104 copy/mL) were 36.5％ (23/63),44.4％ (28/63),42.9％ (27/63) and 54.0％ (34/63),respectively,those in patients with high viral loads (HBVDNA≥1×104 copy/mL) were 8.8％ (5/57),15.8％ (9/57),5.3％ (3/57) and 14.0％ (8/57),respectively.The positive rates of HBxAg and HBxAb were significantly higher in cases with low viral loads than those with high viral loads (x2 =12.869,11.522,22.556 and 20.976,respectively; all P＜0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in the HBeAg positive group were 21.7％ (18/83),30.1％ (25/83),22.9％ (19/83) and 32.5％ (27/83),respectively,while those in the HBeAg negative group were 27.0％ (10/37),32.4％ (12/37),29.7％ (11/37) and 40.5％ (15/37),respectively.No significant difference of HBxAg/HBxAb positive rates between HBeAg positive group and HBeAg negative group was noticed (x2 =0.408,0.064,0.638 and 0.722,respectively; all P＞0.05).Conclusions The antigenicity and specificity of HBV X protein remains similar after the occurrence of A1762T/G1764A double mutant in X gene.It is also found that the positive rates of HBxAg and HBxAb increase with disease progression.HBxAg/HBxAb might be promoting factors for tumorigenesis in chronic HBV infection.HBxAg and HBxAb might have negative influence on HBV replication.

Objective To investigate the effect of phloroglucinol on pregnancy outcome in patients with recurrent implantation failure (RIF).Methods A total of 146 patients with RIF from March 2014 to March 2016 from the reproductive medical center of the Guangxi Zhuang Autonomous Region people's Hospital was randomly divided into two groups,73 cases were included in study group [16 cases of in vitro fertilization and embryo transfer (IVF-ET) and 57 cases of frozen/thawed embryo transfer (FET)].Patients in study group were given intramuscular injection of phloroglucino140mg,two times a day before the transplantation day to three days after transplantation,73 cases without phloroglucinol injection were included as control group.The biochemical pregnancy rate,clinical pregnancy rate,embryo implantation rate,abortion rate,ectopic pregnancy rate,multiple pregnancy rate and live birth rate were compared between two groups.Results The biochemical pregnancy rate in study group of FET was significantly higher than the control group (57.9％ vs 36.8％,P ＜0.05);the biochemical pregnancy rate in study group of IVF-ET was higher than the control group (50％ vs 37.5％,P ＞ 0.05),but there was no significant difference between the study group and control group;compared to the control group,the study group was increased clinical pregnancy rate,implantation rate,live birth rate,and decreased abortion rate (P ＞ 0.05),but the difference was not statistically significant.Conclusions The application of phloroglucinol in women with RIF may improve the biochemical pregnancy rate,especially in FET cycles.

Objective To observe expression of the keratin 10 (K10), c-myc and cox2 in buccal mucous membrane exfoliated cells, and changes of liver function of coal-burning arsenism patients, in order to explorate coal-burning arsenic poison circumference biology symbol. Methods The buccal mucous membrane exfoliated cells were collected from both arsenism patients and normal controls. Real-time PCR was employed to detect the changes of K10, c-myc, cox2 genes expression in these cells. Simultaneously, circumference venous blood was extracted and examinated liver function. Results K10 was found obviously overexpressed in arsenism patients(3.60±0.94) compared to control(1.82±0.68), the difference had statistics significance(t=2.15, P<0.05), c-mye and cox2 did not found obviously changed(c-myc: 3.50±2.77,3.39±2.07; cox2:5.90±1.40,4.73±1.91; t=1.26,1.65, P> 0.05). The serum ALT of patients(25.83±2A5) obviously increased than control(36.86±1735, t=2.55, P<0.05). Conclusions Expression of K10 gene in buccal mucous membrane cells may be regarded as sensitive molecular markers for skin pathologic changes in arsenic patients. The liver is a sensitive target organ of inorganic arsenic.