The diagnostic value of KRAS gene mutation in appendiceal adenocarcinoma / 中国医师杂志

Objective:To analyze the mutation characteristics of Kirsten rat sarcoma virus oncogene homology (KRAS) gene in patients with appendiceal adenocarcinoma and its relationship with the activity of Ras Raf Mitogen activated protein kinase/extracellular regulated protein kinase (MAPK/ERK) signaling pathway.Methods:A total of 41 patients with appendiceal adenocarcinoma who were treated in the Lishui Central Hospital from January 2014 to January 2020 were selected as the observation group, and 50 patients with Appendicitis who were operated at the same time were randomly selected as the control group. Clinical and follow-up data were collected, and the mutation of the KRAS gene in the patient′s tissue was measured using the snapshot method. The expression of key proteins in the MAPK/ERK signaling pathway in cancer tissue was measured using Western blotting (WB) assay. We compared the clinical characteristics and prognosis of patients with KRAS mutation and non KRAS mutation appendiceal adenocarcinoma.Results:The KRAS gene mutation rate in the observation group was higher than that in the control group (41.5% vs 10.0%), and the expression levels of p-ARAF/ARAF, p-MEK1/MEK1, and p-ERK1/ERK1 proteins were also higher than those in the control group. The differences between the groups were statistically significant (all P<0.05). The protein expression levels of p-ARAF/ARAF, p-MEK1/MEK1, p-ERK1/ERK1 in KRAS mutation patients in the observation group were significantly higher than those in non KRAS mutation patients. The proportion of stage IV, positive rates of carcinoembryonic antigen (CEA), carbohydrate antigen (CA)199 and CA125 in KRAS mutation patients were higher than those in non KRAS mutation patients, and the survival time and progression free survival time were shorter than those in non KRAS mutation patients, with statistical significance (all P<0.05). Conclusions:The mutation rate of KRAS in appendix adenocarcinoma is high, and the activation of MAPK/ERK signaling pathway caused by KRAS mutation may play a role in the pathogenesis of appendix adenocarcinoma, which has the value of in-depth research.

Correlation between endoscopic classification and antibiotic resistance of Helicobacter pylori in 5 441 patients with benign gastric diseases / 中华消化杂志

Objective:To analyze the relationship between the gastric diseases classification under endoscopy and infection and antibiotic resistance of Helicobacter pylori ( H. pylori) based on the data of gastric diseases classification under endoscopy, H. pylori isolation and antibiotic resistance. Methods:From January 1, 2015 to December 31, 2019, endoscopic diagnosis and the data of drug susceptibility of H. pylori of 5 441 patients with benign gastric diseases along with gastrointestinal symptoms who visited The First People′s Hospital of Putuo District of Zhoushan were retrospectively collected. The H. pylori infection in patients with different genders and different types of gastric diseases and the drug resistance rates of H. pylori among different antibiotic types were analyzed. Chi-square test was used for statistical analysis. Results:The positive rate of H. pylori in 5 441 patients with benign gastric diseases was 51.37% (2 795/5 441). The drug resistance rates of clarithromycin, levofloxacin, metronidazole, amoxicillin, tetracycline and furazolidone were 26.73% (747/2 795), 43.22% (1 208/2 795), 98.68% (2 758/2 795), 0.32% (9/2 795), 0 and 0, respectively. Among the 2 795 cases of H. pylori-positive patients, only 23 cases (0.82%) were sensitive to all six antibiotics; 1 263 cases (45.19%) were resistant to single antibiotic; 1 072 cases (38.35%) were resistant to combination of two antibiotics, and 433 cases (15.49%) were resistant to combination of three antibiotics. The drug resistance rate to three antibiotics, clarithromycin, levofloxacin and metronidazole simultaneously was the highest (15.31%, 428/2 795). And only four patients (0.14%) were simultaneously resistant to four antibiotics clarithromycin, levofloxacin, metronidazole and amoxicillin. H. pylori infection rate in men was higher than that in women (55.23%, 1 615/2 924 vs. 46.88%, 1 180/2 517), and the difference was statistically significant ( χ2=37.76, P<0.01). However there were no statistically significant differences in the drug resistance rates of six antibiotics between different gender among H. pylori-positive patients (all P>0.05). There was statistically significant difference in the positive rate of H. pylori among patients with different types of gastric diseases ( χ2=909.07, P<0.01). The positive rate of H. pylori was the highest in patients with duodenal ulcer (97.53%, 79/81), and the lowest in patients with bile reflux gastritis (19.79%, 37/187). There were statistically significant differences in the drug resistance rates of metronidazole, clarithromycin, levofloxacin and amoxicillin among 9 types of gastric diseases ( χ2=24.75, 38.62, 80.54 and 7.01, all P<0.01). Conclusions:The positive rate of H. pylori and antibiotic resistance rate are different in patients with different types of gastric diseases. It is necessary to pay attention to the selection of drugs for ulcerative diseases in the process of clinical treatment. Individualized treatment is recommended according to the results of antibiotic susceptibility test.

Causes of oocyte vitrification and its value in assisted reproductive technology / 南方医科大学学报

OBJECTIVE@#To explore the causes of oocyte vitrification and its application in assisted reproduction.@*METHODS@#We retrospectively analyzed the data of 26 patients with 27 cycles of oocyte vitrification cryopreservation undergoing intracytoplasmic sperm injection (ICSI) and embryo transfer between January, 2008 and October, 2018. The causes of oocyte vitrification and the outcomes of ICSI and clinical pregnancy were analyzed.@*RESULTS@#The causes of oocytes vitrification included mainly azoospermia or severe spermatogenesis disorder of the husband, failure to obtain sperms from the husband, failure of the husband to be present on the day of oocyte retrieval and acute diseases of the husband to not allow sperm collection. A total of 274 oocytes were frozen in 27 oocyte retrieval cycles, and 217 eggs were thawed in 19 cycles with a survival rate of 81.11% (176/217). The normal fertilization rate, cleavage rate and high-quality embryo rate was 74.81% (98/131), 89.80% (88/98) and 36.73% (36/98), respectively. Fifteen patients underwent embryo transfer, and the clinical pregnancy rate and live birth rate was 53.33% (8/15) and 33.33% (5/15), respectively. Compared with patients below 35 years of age, the patients aged above 35 years had significantly lower oocyte survival rate after thawing (82.76% 74.42%, =0.211), clinical pregnancy rate (77.78% 16.67%, =0.041) and live birth rate (55.56% 0, =0.044).@*CONCLUSIONS@#Oocytes vitrification can be used as a remedy for infertile couples who fail to provide sperms due to male factors on the day of oocyte retrieval. Vitrification of the oocytes does not significantly affect the fertilization rate or the clinical pregnancy rate. The survival rate of the thawed oocytes is related to the age of the wife, and an age younger than 35 years can be optimal for achieving favorable clinical pregnancy outcomes after oocyte vitrification.

CEP55 may be a potential therapeutic target for non-obstructive azoospermia with maturation arrest / 南方医科大学学报

OBJECTIVE@#To explore the effect of small interfering RNA (siRNA)-mediated CEP55 gene silencing on the proliferation of mouse spermatogonia.@*METHODS@#Six patients with azoospermia diagnosed to have maturation arrest (3 cases) or normal spermatogenesis (3 cases) based on testicular biopsy between January 1 and December 31, 2017 in our center were examined for differential proteins in the testicular tissue using isobaric tags for relative and absolute quantitation (iTRAQ), and CEP55 was found to differentially expressed between the two groups of patients. We constructed a CEP55 siRNA for transfection in mouse spermatogonia and examined the inhibitory effects on CEP55 expressions using Western blotting and qPCR. The effect of CEP55 gene silencing on the proliferation of mouse spermatogonia was evaluated with CCK8 assay.@*RESULTS@#In the testicular tissues from the 6 patients with azoospermia, iTRAQ combined with LC/MS/MS analysis identified over two hundred differentially expressed proteins, among which CEP55 showed the most significant differential expression between the patients with maturation arrest and those with normal spermatogenesis. The cell transfection experiment showed that compared with the cells transfected with the vehicle or the negative control sequence, the mouse spermatogonia transfected with CEP55 siRNA showed significantly lowered expressions of CEP55 mRNA and protein ( < 0.05) and significantly decreased proliferation rate as shown by CCK8 assay ( < 0.05).@*CONCLUSIONS@#CEP55 may play a key role in spermatogenesis and may serve as a potential therapeutic target for non-obstructive azoospermia with maturation arrest.

Feasibility study of transplantation of penile corpus cavernosum and major pelvic ganglion in renal subserous region / 北京大学学报(医学版)

Objective:To study the feasibility of transplantation of normal rat penile corpus cavernosum and major pelvic ganglion (MPG)into the renal subserous region of a Nu /Nu mouse based on allograft technology.Methods:Penile corpus cavernosum and MPG,harvested from Sprague-Dawley (SD)rats under sterile condition,were transplanted underneath the kidney capsule of Nu /Nu mice through the mi-crosurgery instruments and surgery microscope.The histopathologic changes and cellular proliferation in the transplanted penile corpus cavernosum and MPG were then analyzed at the end of 1week and 4 weeks after transplantation.Histological staining and immunohistochemical staining were used to evaluate the main outcome measures.Results:After 1 week,the tissue morphology of the transplanted corpus caverno-sum underneath the kidney capsule of Nu /Nu mice was consistent with normal penile corpus cavernosum, and blood could be observed in the penis cavernous sinus of the graft;after 4 weeks,the mophorlogy of the tranplanted corpus cavernosum near the kidney was consistent with normal penile corpus cavernosum, while fibrosis was noteworthy in the graft away from the kidney,but blood could still be seen in the penis cavernous sinus.After 1 week,the tissue morphology of the transplanted MPG was consistent with normal MPG,multiple islet-like cell clusters could be seen in the transplanted MPG in the renal subserous re-gion,and angiogenesis could be observed near the kidney;after 4 weeks,a network of blood vessels was clearly visible away from the kidney,and islet-like cell clusters were still clearly observed in the trans-planted MPG.In addition,ki67 positive cells were observed in the transplanted penile corpus cavernosum and MPG after 4 weeks of transplantation,which indicated that there was still cell proliferation activity in the grafts.Conclusion:The transplanted corpus cavernosum and MPG underneath the kidney capsule of Nu /Nu mice could survive at least 4 weeks.Moreover,the inner structure of the transplanted corpus ca-vernosum and MPG was close to the normal tissue.The underlining mechanism may be related to the lo-cal microenvironment underneath the kidney capsule of Nu /Nu mice and the neovascularization in the transplanted grafts.

Low-Intensity Shock Wave Therapy and Its Application to Erectile Dysfunction

Although phosphodiesterase type 5 inhibitors (PDE5Is) are a revolution in the treatment of erectile dysfunction (ED) and have been marketed since 1998, they cannot restore pathological changes in the penis. Low-energy shock wave therapy (LESWT) has been developed for treating ED, and clinical studies have shown that LESWT has the potential to affect PDE5I non-responders with ED with few adverse effects. Animal studies have shown that LESWT significantly improves penile hemodynamics and restores pathological changes in the penis of diabetic ED animal models. Although the mechanisms remain to be investigated, recent studies have reported that LESWT could partially restore corpus cavernosum fibromuscular pathological changes, endothelial dysfunction, and peripheral neuropathy. LESWT could be a novel modality for treating ED, and particularly PDE5I non-responders with organic ED, in the near future. However, further extensive evidence-based basic and clinical studies are needed. This review intends to summarize the scientific background underlying the effect of LESWT on ED.