Objective To discuss risk factor and operation guard in dissection of cholecyat from its bed during laparoseopic cholecystectomy.Methods The clinical data of 490 cases of laparoscopic cholecystectomy dissect cholecyst from its bed by traditional method during were retrospectively analyzed and simultaneously perspective study on 500 cases of improved laparoscopic cholecyatectomy dissecting cholecyst from its bed.Results There are 3 cases in hemorrhea due to injuring bed brancha of arteria cystica posterior branch,2 cases in bed brancha of arteria cystica posterior branch haemorrhage,5 cases in postoperative bile leak,2 cases in variant right hepatic portal vein injury,1 case in variant right hepatic duct in the 490 cases of laparoscopic choleeystectomy by traditiorhI method.Using our new technique exposed,and then ligated 39.6%(198/500)bed brancha of arteria cystica posterior branch,11.2% (56/500)bed brancha of venae hepaticae intermediate almost in fight side of cholecyst bed,2.2%(11/500)dilated aberrant bile-duct duct.We separated and then protected 0.6%(3/500)unusual right hepatic duct being exposed,0.4%(2/500)right liver branch of portsl vein.Conclusion There are some larvate risk in isolation of choleeyst from its bed during laparoseopic cholecystectomy.Unexpected injury happen during LC,and it ought tO be open cholecysteetomy The improved method provides a safe,efficiency operative procedure.

Objective To introduce a two-stage operative method to treat ma lu nion after the distal femoral fracture. Methods From Aug. 1992 to Aug. 2000, 12 patients underwent osteotomy and secondary fixation due to malunion after the di stal femoral fracture. There were 10 closed fractures and 2 open ones. The prima ry treatment was conservative for 2 patients, and operative for 10. We first per formed the osteotomy along the primary fracture line where the deformity was the most severe. When the roentgenogram confirmed that the deformity had been corre cted satisfactorily, a secondary operation of ORIF (open reduction with internal fixation) was performed. Results On average, all the 12 patients gained bone un ion 5.1 months postoperatively. The mean follow-up duration was 7.2 years (rang e: 3.5 to 11.5 years). None of the patients was afflicted by infection or loosen ing of the internal fixator. According to the Schatzker scoring scale system, al l the patients were evaluated as poor preoperatively, but as excellent in 7, goo d in 4 and fair in 1 at the last follow-up. Conclusion Osteotomy and secondary fixation is not only simple and accessible but also can lead to confirmed clinic al results. However, it is still very important to reduce and fix the acute frac tures stably to avoid malunion.

Objective To investigate the role of CD4+CD25+Foxp3 regulatory T cells in chronicity of hepatitis B and viral clearance of hepatitis B virus(HBV).Methods Nineteen patients with chronic active hepatitis B(CAH).21 HBV carriers(AsC)and 12 patients with resolved HBV infection and 1 5 healthy controls were enrolled.The frequency and phenotype of peripheral CD4+CD25+Foxp3+ T cells were detected by flow cytometry.CD4+CD25+T cells were sorted by magnetic-activated cell sorting(MACS)assay.Level of Foxp3 mRNA in CD4+CD25+T cells was examined by real time polymerase chain reaction(PCR)assay.The data were analyzed by one-way ANoVA or nonparametric statistics.Results Both frequencies of CD4+CD25+Foxp3+T cells and levels of Foxp3 mRNA in CD4+CD25+T ceils in patients with CAH or AsC were significantly higher than those in healthy controls Or resolved HBV infection(F=6.8,F=3.72,respectively;both P<0.05).Accumulation of Foxp3+T cells in liver tissue of CAH patients was higher than that of healthy controls,while that in AsC was lower than CAH.The frequency of CD4+CD25+Foxp3+T cells of hepatitis B e antigen(HBeAg)positive patients(including CAH and AsC)was significantly higher than that of HBeAg negative patients(t=2.3,P<0.05),and that of antFHBe negative patients were significantly higher than anti-HBe positive patients(t=2.4,P<0.05).Furthermore,the frequency of CD4+CD25+Foxp3 regulatory T cells was positively correlated with serum HBV DNA level of patients with chronic hepatitis B(r=0.56,P<0.01).Conclusion The findings have important implication in the understanding of the role of CD4'CD25'regulatory T cells in chronicity and viral clearance in HBV infection.

OBJECTIVE:To study the effect and thepossible mechanism of low molecular weight heparin(LMWH)on immunocytes' adhesion to fibroblast-like synovocytes(FLS)isolated from patients with rheumatoid arthritis(RA)and to study LMWH'S possible anti-inflammatory effect on RA.METHODS:LMWH's interference on the adhesion of peripheral blood monouclear cells(PBMC)isolated from healthy volunteers to FLS of RA patients was determined by quantitative counting using flow cytometry.The expression of CYR61 in the in vitro cultured FLS of RA patients was detected using real-time PCR technology.RESULTS:When FLS culture system was added with PBMC,PBMC were obviously found to adhere to FLS,but the number of adhered PBMC decreased after LMWH treatment,which manifested as increase of deciduous PBMC,increased more with the increase of LMWH dose.There was a high expression of CYR61 in synovium tissue in RA patients.CONCLUSION:LMWH inhibited the adheresion of PBMC to FLS from RA patients in a dose-dependent manner,which might be attributed to its competitive combination with heparin sulfate sites on CYR61.

Objective To comparatively analyze the advantages and disadvantages between a double immune-diffusion assay and a rate nephelometer analysis for the detection of antigen activity of pneu -mococcal capsular polysaccharides .Methods The antigen activity of pneumococcal capsular polysaccharides of serotypes 1,6B,9V,10A,14 and 19A from four manufacturers and ATCC were analyzed by a double im-mune-diffusion assay and a rate nephelometer analysis , respectively .The effects of antiserum samples and gain values on the rate response value were evaluated .Results The sample 4 of type 9V showed no antigeni-city with a rate response value similar to that of negative control as indicated by both tests .However ,the pre-cipitation lines and the rate response values presented by other polysaccharide samples differed in a wide range.Results of the rate nephelometer analysis were not affected by the anti -serum samples from different sources and the gain values .Conclusion The rate nephelometer analysis could quantitatively analyze the antigen activity of pneumococcal capsular polysaccharides .

0.05).CONCLUSION: The high expression of GPC3 mRNA in hepatocellular carcinoma is independent to the gene mutation,and to the expression of P53 and PCNA protein.

OBJECTIVETo confirm the diagnosis of a Wolf-Hirschhorn syndrome by family study using both cytogenetic and molecular genetic techniques.

METHODG-band karyotyping was performed for all the 6 members in the family. Multiplex ligation-dependent probe amplification (MLPA) was used to detect the chromosome abnormality for the proband, his father and brother. Microarray comparative genomic hybridization (Array-CGH) was carried out to map the exact chromosomal breakpoints for the proband.

RESULTThe proband presented with a typical face, delayed growth and hypotonia in Wolf-Hirschhorn syndrome. His G-band karyotype was 46, XY, der(4)t(4;8) (p16.2; p23.1)pat. MLPA showed 4pter loss and 8pter gain. Array-CGH revealed an XY male with a 3.781 Mb deletion of 4p16.3-p16.2 and a 6.760 Mb duplication of 8p23.3-p23.1. The proband's brother has mental retardation and skeletal abnormalities. His G-band karyotype was 46, XY, der(8)t(4;8)(p16.2;p23.1)pat. MLPA showed 4pter gain and 8pter loss. The proband's father had normal phenotype with a balanced translocation of 46, XY, t(4;8)(p16.2;p23.1)pat. MLPA showed a normal result. The proband's grandfather showed a normal phenotype with a balanced translocation 46, XY, t(4;8)(p16.2;p23.1). The other members in the family showed normal phenotypes with normal karyotypes.

CONCLUSIONThe proband has features of Wolf-Hirschhorn syndrome with partial monosomy 4p and partial trisomy 8p. The proband's brother has a partial trisomy 4p and partial monosomy 8p. The derived chromosomes are inherited from paternal balanced translocation t(4;8)(p16.2;p23.1).

Abnormalities, Multiple ; genetics ; Adult ; Chromosome Deletion ; Chromosomes, Human, Pair 4 ; genetics ; Chromosomes, Human, Pair 8 ; genetics ; Comparative Genomic Hybridization ; Female ; Humans ; Infant ; Karyotyping ; Male ; Multiplex Polymerase Chain Reaction ; methods ; Oligonucleotide Array Sequence Analysis ; Pedigree ; Phenotype ; Translocation, Genetic ; Trisomy ; Wolf-Hirschhorn Syndrome ; diagnosis ; genetics

AIM: To analyze the diagnostic value of optical coherence tomography angiography(OCTA)in evaluating the parameters of foveal avascular zone(FAZ)in early diabetic retinopathy(DR).METHODS: Retrospective study. A total of 209 cases(209 eyes)of type 2 diabetes mellitus(T2DM)with DR admitted to our hospital from June 2019 to December 2022 were selected as DR group. The DR group was divided into three groups, with 115 cases(115 eyes)in mild group, 54 cases(54 eyes)in moderate group, and 40 cases(40 eyes)in severe group according to stage. Another 100 cases(100 eyes)of T2DM patients without DR were selected as No-DR group, and 70 cases(70 eyes)of healthy people were selected as control group for physical examination at the same time, all of whom underwent OCTA examination. The DR group was enrolled according to the disease degree, one eye was randomly taken for the study if the degree in both eyes was the same, while the control group and the No-DR group were randomly selected for one eye to be included in the study. The perimeter of the foveal avascular area(PERIM), FAZ transverse diameter, FAZ vertical diameter, FAZ area, macular fovea retinal thickness(MFRT), acircularity index(AI), full layer retinal blood flow density within a range of 300 μm around the FAZ(FD-300), and changes in FAZ vascular density(VD)levels among different groups of subjects were compared. Furthermore, Pearson correlation analysis was used to determine the correlation between general data and FAZ related indicators, and receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of macular FAZ indicators for DR.RESULTS: The levels of FBG, MFRT, FAZ area, and PERIM in patients with mild, moderate, and severe DR were significantly higher than those in the No-DR group and the control group, while the levels of AI and VD were significantly lower than those in the No-DR group(all P<0.05); patients in the No-DR group, mild DR group, moderate DR group, and severe DR group had significantly higher cystatin C, FAZ transverse diameter, and FAZ vertical diameter than the control group, while FD-300 vascular density was significantly lower than the control group(all P<0.05); the MFRT was positively correlated with age(r=0.295, P=0.001); The AI and VD was negatively correlated with age(r=-0.296, -0.235, all P<0.05); the area under the curve(AUC)of MFRT, FAZ area, PERIM, AI, and VD for the diagnosis of DR were 0.745, 0.738, 0.696, 0.792, and 0.847, respectively.CONCLUSION:The structure and microcirculation of FAZ can be changed in DM patients, and the related parameters of FAZ have certain value in the diagnosis of early DR.

Di-(2-ethylhexyl) phthalate (DEHP) is currently one of the most widely used plasticizers, widely found in all kinds of items, such as children’s toys and food packaging materials, but also added to wallpaper, cable protective agents and other building decoration materials. DEHP is toxic and absorbed by the human body through respiratory tract, digestive tract and skin contact, which can cause damage to multiple systems, especially the male reproductive system, and testis is an important target organ. Oxidative stress injury is the core mechanism of spermatogenesis disorder caused by DEHP. DEHP exposure can cause oxidative stress or reactive oxygen species (ROS) increase in germ cells, and on this basis, promote cell apoptosis or cause excessive autophagy. The toxicity of DEHP to Leydig cells is mainly to interfere with the synthesis of steroid hormones. For Sertoli cells, ferroptosis and destruction of the blood-testis barrier are common injury mechanisms. In addition, gene methylation caused by DEHP not only affects the spermatogenic process, but also has epigenetic effects on offspring. In this paper, we reviewed the pathological damage, germ cell toxicity and epigenetic effects of DEHP on testis, and focused on the damage and molecular mechanism on testicular spermatogenic cells, Leydig cells and Sertoli cells. Future research is required to elucidate the body’s clearance mechanism and treatment plan after exposure to DEHP and whether DEHP will damage the function of myoid cells. It is hoped that this can provide new ideas for prevention and treatment of male reproductive disorders resulting from long-term exposure to plastic products.

Objective: To evaluate the relationship between the hippocampal neuron-protective mechanism of hydrogen in a rat model of oxygen-glucose deprivation and restoration (OGD/R) and mitochondrial autophagy. Methods: Hippocampal neurons isolated from healthy Sprague-Dawley rats (24 h after birth) were cultured in vitro, seeded in polylysine-coated 6-well plates at a density of 7×10⁵ cells/well and then divided into 5 groups (n=30 each) using a random number table method: control group (C group), OGD/R group, OGD/R+ H₂ group, OGD/R plus 3-methyladenine (3-MA) group (OGD/R+ 3-MA group), and OGD/R plus H₂ plus 3-MA group (OGD/R+ H₂+ 3-MA group). The cells were cultured for 24 h in normal culture atmosphere (75%N₂-20%O₂-5%CO₂) in group C, and cells were subjected to oxygen-glucose deprivation for 2 h followed by O₂-glucose supply for 24 h to establish the model of OGD/R injury in OGD/R, OGD/R+ H₂, OGD/R+ 3-MA and OGD/R+ H₂+ 3-MA groups.The cells were cultured for 24 h in a hydrogen-rich incubator (60% H₂-10% O₂-5% CO₂-25% N₂) after establishing the model in group OGD/R+ H₂.Autophagy inhibitor 3-MA 10 mmol/L was added, and then cultured for 24 h in normal culture atmosphere after establishing the model in group OGD/R+ 3-MA.Autophagy inhibitor 3-MA 10 mmol/L was added, and then cultured for 24 h in hydrogen-rich incubator after establishing the model in group OGD/R+ H₂+ 3-MA.The cell survival rate was measured using MTT assay.DCFH-DA fluorescent probe was applied for determination of reactive oxygen species (ROS) activity.The mitochondrial membrane potential was measured using a JC-10 assay kit.The neuronal apoptosis was detected by flow cytometry, and apoptosis rate was calculated.The expression of mitophagy-related protein microtubule-associated protein 1 light chain 3 (LC3), PINK1 and Parkin was determined by Western blot, and LC3Ⅱ/LC3Ⅰ ratio was calculated. Results: Compared with group C, the cell survival rate and MMP were significantly decreased, the apoptosis rate and ROS activity were increased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were increased in OGD/R and OGD/R+ H₂ groups (P<0.05). Compared with group OGD/R, the cell survival rate and MMP were significantly increased, the apoptosis rate and ROS activity were decreased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were increased in group OGD/R+ H₂(P<0.05), and the cell survival rate and MMP were significantly decreased, the apoptosis rate and ROS activity were increased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were decreased in group OGD/R+ 3-MA (P<0.05). Compared with group OGD/R+ H₂, the cell survival rate and MMP were significantly decreased, the apoptosis rate and ROS activity were increased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were decreased in OGD/R+ 3-MA and OGD/R+ H₂+ 3-MA groups (P<0.05). Conclusion Hippocampal neuron-protective mechanism of hydrogen against OGDR injury is related to promoting mitochondrial autophagy in rats.