Objective To study the indication,method and value of choledochal exploration through cystic duct.Methods The cystic duct was cut 0.5～1 cm from choledochus and its edge was raised with a forceps and a bakes dilator was inserted to probe the bill duct.If the dilator can not pass the choledochus,palpation around the point of dilator would help to distinguish the stone tumor or narrow.Results In 51 patients with cholecystitis and cholelithiasis,36 were explorated,of which 6 had the explorative indications,4 were postive(66.6%);in 30 without the indications,3 were positive(10%).The total positvity was 19.4%.Conclusions This method is less traumatic,convenient and able to find choledochal disease accurately.The exploration would be done if the cystic duct is over 0.3 cm in diameter.

[Summary] Neuroischemic diabetic foot ulcer ( NDFU) is characterized by infection, ulceration of deep tissues, neurological abnormalities, and various degrees of peripheral vascular disease in the lower limbs. The patients often have multiple risk factors such as older, longer duration, cardiovascular disease. The treatment is very difficult. The prognosis depends on the severity of complications, tissue range of infections, and the peripheral vascular disease. In this article, the treatment process of an old inpatient with NDFU and severe complications was reviewed and to propose a standard pathway for its management.

Objective To observe the influences of uncoupling protein 2 (UCP-2) rs660339 variants transfection on cell proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods Two UCP-2 green fluorescent protein (GFP) lentivirus constructs were created with the rs660339 locus carried C or T (UCP-2C or UCP-2T), respectively. HUVEC were cultured after lentiviral infection of UCP-2C or UCP-2T. The expression of UCP-2C or UCP-2T was detected with real time polymerase chain reaction. Cell proliferation and cell apoptosis were compared among negative control (NC) group, UCP-2T group and UCP-2C group using CCK-8 cell viability and flow cytometry. Western blot and immunostaining were employed to examine the expression of Bcl-2 gene. Results The lentivirus constructs were successfully created.>80%of the transfected cells were found to express GFP under fluorescent microscope. The mRNA levels of UCP-2 gene were significantly increased (F=29.183, P=0.001) in the UCP-2T group and UCP-2C group. The CCK-8 assay revealed that on day two (F=15.970, P=0.004), day three (F=16.738, P=0.004), day four (F=5.414, P=0.045) post-infection, UCP-2T and UCP-2C group showed significantly greater proliferation than the NC cells. The apoptotic rate in the UCP-2T and UCP-2C group was significantly lower than NC group (F=277.138, P=0.000), and the apoptotic rate of UCP-2T was significantly lower than that of UCP-2C (P=0.003). The protein levels of Bcl-2 in the UCP-2T and UCP-2C group were significantly greater than that in the NC group (F=425.679, P=0.000), and the Bcl-2 expression of UCP-2T was greater than that of UCP-2C (P=0.002). The Bcl-2 density in the UCP-2T and UCP-2C group were greater than that in the NC group (F=11.827, P=0.008), while there was no difference between UCP-2T and UCP-2C group (P=0.404). Conclusion The variants of UCP-2 rs660339 may influence HUVEC proliferation and apoptosis, and UCP-2T showed a stronger effect of inhibiting apoptosis than UCP-2C.

This exploratory study attempted to establish the rules in diagnosis and treatment of diabetic foot, through interpretation and comparison of the guidelines for diabetic foot from domestic with international ones. The document provided comparison between Guidelines of International Working Group on Diabetic Foot (IWGDF) and Chinese Diabetes Society (CDS) in 2019, related to diabetic foot disease on: methodology, content, prevention, offloading, peripheral artery disease (PAD), infection, wound healing interventions, and classification of diabetic foot ulcers. Prevention of ulcers in persons with diabetes foot is very important, and a non-removable offloading device is the first-choice of offloading treatment; Surgical indications and reasonable treatment should be mastered in PAD; Different anti-infection treatments, including surgical debridement, should be used base on the severity of foot infection; There are lots of treatments to improve healing, however the Grand Standard of medical evidence is not very high; There are a larger number of proposed classifications and scoring systems for diabetic foot, but none of them could cover all the needs of diagnosis and treatment. Thus, the principles outlined have to be adapted or modified by our health care professionals, based on local circumstances, to develop a standardizated procedure in diagnosis and treatment of diabetic foot.

Objective:To investigate the mechanism of maggot debridement therapy (MDT) in promoting wound angiogenesis in patients with diabetic foot ulcer (DFU).Methods:(1) From June 2018 to June 2019, the patients admitted to Nanjing Junxie Hospital who met the inclusion criteria were recruited, including 12 DFU patients given MDT for three days [6 males and 6 females, aged (56±12) years] and 12 acute trauma patients without diabetes mellitus [6 males and 6 females, aged (53±10) years], who were enrolled into DFU group and non-diabetic trauma group respectively. Before and after application of MDT, the wound characteristics of patients in DFU group were observed and the wound tissue samples were taken. The wound tissue in non-diabetic trauma group was taken at patient′s first visit before debridement. The expression of angiogenesis marker CD31 in the wound tissue of patients in DFU group was detected by immunohistochemistry before and after application of MDT. Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) were used respectively to detect the protein and mRNA expressions of fatty acid synthase (FAS) in wound tissue of patients in DFU group before and after application of MDT and in non-diabetic trauma group before debridement. (2) Human umbilical vein endothelial cells (HUVECs) were cultured in endothelial cell culture medium containing 10% fetal bovine serum. The 3rd to 6th passages of cells in logarithmic growth phase were used in the following experiments. Excretions/secretions (ES) were extracted from 3-day-old sterile Lucilia sericata larvae for subsequent experiments. Three batches of cells were divided into phosphate buffer solution (PBS) control group, high glucose alone group, high glucose+ 5 μg/mL maggot ES group, and high glucose+ 10 μg/mL maggot ES group, which were treated with PBS, glucose in final molarity concentration of 20 mmol/L, glucose in final molarity concentration of 20 mmol/L+ maggot ES in final mass concentration of 5 μg/mL, and glucose in final molarity concentration of 20 mmol/L+ maggot ES in final mass concentration of 10 μg/mL respectively. The total volume of reagents in each group was the same. After 48 hours of culture, Western blotting, real-time fluorescent quantitative RT-PCR and immunofluorescence method were used to detect the protein and mRNA expressions of FAS in each batch of cells and the expression and localization of FAS protein in cells respectively. The number of samples for mRNA expression was 3. (3) Two batches of cells were divided into small interference RNA (siRNA) alone group, siRNA control+ maggot ES group and siRNA-FAS+ maggot ES group, which were transfected with 100 μmol/L (final molarity concentration) insignificant control siRNA, insignificant control siRNA, and siRNA-FAS for 4-6 h respectively, and then they were routinely cultured for 24 h with PBS added, maggot ES in final mass concentration of 10 μg/mL, and maggot ES in final mass concentration of 10 μg/mL respectively. The total volume of reagents in each group was the same. One batch of cells was used for scratch test, the scratch width was observed at 24 hour after scratching to detect the cell migration ability; one batch of cells was subjected to tube forming experiment, and the formation of cell tubules was observed after 24 hours of culture. The number of samples was 3 in scratch test and tube forming experiments. Data were statistically analyzed with t test, one-way analysis of variance, least significant difference test, analysis of variance for repeated measurement, and Bonferroni method. Results:(1) Compared with those before application of MDT, fresh granulation tissue significantly increased and necrotic tissue decreased obviously in wound, and the expression of CD31 significantly increased in wound tissue of patients in DFU group after application of MDT. The expression of FAS protein in wound tissue of patients in DFU group before application of MDT was significantly lower than that in non-diabetic trauma group before debridement, and the expression of FAS protein in wound tissue of patients in DFU group after application of MDT was significantly higher than that before application of MDT. The expression of FAS mRNA in wound tissue of patients in DFU group before application of MDT was 1.00±0.17, which was significantly less than 3.87±1.02 in non-diabetic trauma group before debridement ( t=9.808, P<0.01). The expression of FAS mRNA in wound tissue of patients in DFU group after application of MDT was 1.85±0.31, which was significantly higher than that before application of MDT ( t=-10.853, P<0.01). (2) After 48 hours of culture, Western blotting detection showed that the expression of FAS protein in cells in high glucose alone group was significantly less than that in PBS control group, and the expressions of FAS protein in cells in high glucose+ 5 μg/mL maggot ES group and high glucose+ 10 μg/mL maggot ES group were significantly higher than the expression in high glucose alone group. Real-time fluorescent quantitative RT-PCR determination showed that the expression of FAS mRNA in cells in high glucose alone group was 0.392±0.073, which was significantly lower than 1.000±0.085 in PBS control group ( P<0.01); there was statistically significant difference between the expression of FAS mRNA in cells in high glucose+ 5 μg/mL maggot ES group (0.561±0.047) and that in high glucose+ 10 μg/mL maggot ES group (0.687±0.013) ( P<0.05), both of which were significantly higher than the expression in high glucose alone group ( P<0.01). The results of immunofluorescence detection showed that FAS protein was mainly located in the cytoplasm of cells in each group, and its expression was similar to that detected by Western blotting. (3) At 24 hour after scratch, the uncured widths of cell scratch in siRNA control+ maggot ES group and siRNA-FAS+ maggot ES group were significantly narrower than the uncured width in siRNA alone control group ( P<0.01), and the uncured width of cell scratch in siRNA-FAS+ maggot ES group was significantly wider than that in siRNA control+ maggot ES group ( P<0.01). After 24 hours of culture, the numbers of tubules in siRNA+ maggot ES group and siRNA-FAS+ maggot ES group were significantly more than the number in siRNA alone control group ( P<0.05 or P<0.01), and the number of tubules in siRNA-FAS+ maggot ES group was obviously less than that in siRNA control+ maggot ES group ( P<0.05). Conclusions:MDT up-regulates the expression of FAS through maggot ES, which promotes the activity of vascular endothelial cells, thus promoting the wound angiogenesis in patients with DFU.

Ginsenoside Compound K(CK)is a kind of protopanaxadiol ginsenosides,which exerts antitumor effects on various cancers,such as lung cancer,liver cancer and breast cancer.Pharmacological studies have proved that CK induces tumor cell cycle arrest and apoptosis,as well as regulates tumor cell autophagy and inhibits tumor metastasis by regulating multiple signaling pathways(AMPK/mTOR and PI3K/Akt et cetera).However,as an intestinal metabolite of natural protopanaxadiol ginsenosides,CK cannot be directly extracted from the ginseng plant.Therefore,biotransformation from natural ginsenosides such as ginsenoside Rb1 or biosynthesis are utilized to obtain CK.Biotransformation of CK uses enzymes or microorganisms to transform different ginsenosides or their structural analogs into CK;biosynthesis of CK cultures cell factories and biosynthetic enzymes to synthesize glucose and other simple compounds into CK.In this review,we systematically summarized the research progress in biopreparation and antitumor mechanism of CK in recent years,with the aim of providing evidence for the future development of CK as a clinical anti-tumor candidate drug or an adjunctive drug.