Acute kidney injury (AKI) is one of the most common and severe nephropathy syndromes in clinical practice and also one of the most common serious complications after organ transplantation, with high incidence and fatality. Iron is an essential trace element in the body. Ferroptosis is a form of programmed cell death induced by the accumulation of iron-mediated lipid peroxidation, and its occurrence is closely related to iron metabolism, lipid metabolism, amino acid metabolism and multiple signaling pathways. Recent studies have shown that ferroptosis plays a key role in the occurrence and development of AKI and provides therapeutic targets for AKI. This article summarizes the regulatory mechanism of ferroptosis and its role in AKI, as well as the compounds that play an important role in the prevention and treatment of AKI by inhibiting ferroptosis, providing new ideas for the future treatment and research of AKI.

The fcl gene encodes GDP-fucose synthase, which catalyzes two-step differential isomerase and reductase reactions in the synthesis of GDP-L-fucose from GDP-D-mannose. It also participates in the biosynthesis of amino sugar and ribose sugar, and is one of the key enzymes to regulate the metabolism of sugar and nucleotides in organisms. The presence of fcl gene in Saccharopolyspora pogona was found through sequencing result of genome. The mutant S. pogona-fcl and S. pogona-Δfcl were constructed by gene engineering technology. The results showed that the gene had an effects on growth and development, protein expression and transcriptional level, insecticidal activity, and biosynthesis of butenyl-spinosyn of Saccharopolyspora pogona. The results of HPLC analysis showed that the yield of butenyl-spinosyn in S. pogona-Δfcl was 130% compared with that in S. pogona, which reduced by 25% in S. pogona-fcl. The results of determination of insecticidal activity showed that S. pogona-Δfcl had a stronger insecticidal activity against Helicoverpa armigera than that of S. pogona, while the S. pogona-fcl had a lower insecticidal activity against Helicoverpa armigera compared with S. pogona. Scanning electron microscopy (SEM) was used to observe the morphology of the mycelia. It was found that the surface of the S. pogona-Δfcl was wrinkled, and the mycelium showed a short rod shape. There was no significant difference in mycelial morphology between S. pogona-fcl and S. pogona. Aboved all showed that deletion of fcl gene in S. pogona hindered the growth and development of mycelia, but was beneficial to increase the biosynthesis of butenyl-spinosyn and improve insecticidal activity. Whereas the fcl gene over-expression was not conducive to the biosynthesis of butenyl-spinosyn and reduced their insecticidal activity. SDS-PAGE results showed that the difference of protein expression among the three strains was most obvious at 96 hours, which was identified by real-time fluorescence quantitative polymerase chain reaction, the results showed that there were significant differences of related genes in transcriptional levels among the three strains. Based on the results of the study, a network metabolic control map was constructed to analyze the effect of fcl gene on growth and the regulation pathway of butenyl-spinosyn biosynthesis, which provided an experimental basis for revealing the regulation mechanism of butenyl-spinosyn biosynthesis and related follow-up studies.
Bacterial Proteins ; Genetic Engineering ; Insecticides ; Macrolides ; Saccharopolyspora

Objective To analysis the change of hip joint in healthy infants by ultrasound,and establish the normal reference value of the developmental dysplasia of the hip(DDH). Methods A total of 8 000 healthy infants from 0 to 24 weeks were collected from the Multi-center study of 10 children′s medical centers. Among them,3 855 infants(2 065 females and 1 790 males) with complete data and follow-up were included in this study. All subjects were divided into 6 groups ( ＜4,4～7,8～11,12～15,16～19 and≥20 weeks group). α angle,femoral head length and width,femoral head coverage ratio by acetabulum ( FHC) were measured in the coronal view on the neutral position;distance from pubis to femoral head ( P-H) and distance from ischium to femoral head ( I-H ) were measured in the transverse view on neutral position;distance from femoral head topubis ( H-P) was measured in the posterolateraltransverse view on the flexion position. The results of each group changes with age were analysised. Results ① The α angle of healthy infants from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05),but there was no significant difference between 16～19 and ≥20 weeks group( P ＞0.05). ②The femoral head length and width of all age groups were increased with age,the difference among all the groups was statistically significant( all P ＜0.05). ③ FHC from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05) except between 16～19 and ≥20 weeks group( P ＞0.05). ④ The P-H and I-H in all age groups showed no statistically significant ( all P＞0.05). ⑤The H-P of all age groups were increased with age,the difference between the groups were statistically significant(all P ＜0.05).Conclusions The development of hip joints have the certain regular developmental pattern in healthy infants less than 5 months of birth and are relatively constant after birth more than 5 months. The ultrasound normal reference value of the hip joints can be used for the early diagnosis of the DDH.

This study was aimed to observe the clinical effect of low frequency negative pressure meridian viscera regulating sub-health.According to the diagnostic criteria,34 cases of sub healthy subjects were randomly selected.The low frequency negative pressure meridian viscera regulating method was used.By analyzing the cupping spot color feedback conditioning information of the sub healthy subjects and conditioning before and after the synthesis of the four diagnostic methods of digital and quantitative,its effect was evaluated.After 4 weeks of continuous adjustment,34 cases of sub healthy subjects,compared with preconditioning,cupping spot color of the subjects was shallow or disappeared (P ＜ 0.01);after adjusting,the proportion of light red tongue and thin white tongue coating increased obviously (P ＜ 0.05),the proportion of ecchymosis tongue was decreased compared to precoditioning (P ＜ 0.05);after adjustment,core symptoms such as back pain,neck pain,anxiety and irritability,were alleviated or disappeared;after adjustment,the pulse string tightness decreased significantly (P ＜ 0.01).It was concluded that the low frequency negative pressure meridian viscera regulation can significantly improve discomfort symptoms of sub healthy subjects and promote therecovery of tongue,pulse and cupping spot color in sub healthy subjects.This method is worth popularizing and further exploring.

OBJECTIVETo observe the effects of close-to-bone needing combined with electroacupuncture (EA) on cartilage collagen type Ⅱ/discoidin domain receptor/matrix metalloproteinase 13 (collagen type Ⅱ/DDR2/MMP 13) signaling pathway in rabbits with knee osteoarthritis (KOA), and to explore the possible action mechanism of this method on repair of extracellular matrix of knee cartilage.

METHODSForty New Zealand white rabbits were randomly assigned into a normal group (10 rabbits) and a model establishing group (30 rabbits). The Hulth-Telhag technique was applied to establish the model of KOA, and X-ray was used for outcome assessment. The rabbits with successful modeling were randomly assigned into a model group, a close-to-bone needing group, a regular acupuncture group, 10 rabbits in each one. The rabbits in the close-to-bone needing group were treated with close-to-bone needing and EA; the rabbits in the regular acupuncture group were treated with regular acupuncture and EA. "Neixiyan" (EX-LE 4), "Dubi" (ST 35), "Yinlingquan" (SP 9), "Zusanli" (ST 36) and "Liangqiu" (ST 34) were selected in the two groups. The intervention was given for 20 min, once a day; the intervention of 5 days made 1 session, 2 days as the interval and totally 4 sessions were given. Rabbits in normal and model group were immobilized without any treatment. After the treatment, western blotting method was applied to evaluate the expression of DDR2 and collagen type Ⅱ; the activity of collagen type Ⅱ, DDR2 and MMP 13 was assessed by immunohistochemistry method; the mRNA expression of DDR2 and MMP 13 was determined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal group, the activity expression of collagen type Ⅱ were significantly reduced in the other groups (all<0.01),while the activity and mRNA expression of DDR2 and MMP 13 were notably increased (all<0.01). Compared with the model group, the activity expression of collagen type Ⅱ in the close-to-bone needing group and regular acupuncture group were increased (all<0.01), while the activity and mRNA expression of DDR2 and MMP 13 were reduced (all<0.01). Compared with the regular acupuncture group, the activity and mRNA expression of MMP 13 and DDR2 in the close-to-bone needing group were reduced (all<0.01), while the activity expression of collagen type Ⅱ were increased (<0.01).

CONCLUSIONSThe close-to-bone needing combined with EA and regular EA could both promote the repair of knee cartilage, where closing-to-bone needing combined with EA shows a superior efficacy. The mechanism may be associated with the blocking effect of collagen type Ⅱ/DDR2/MMP13 signaling pathway and the inhibiting effect of degradation in extracellular matrix of cartilage.

Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
Genetic Engineering ; Macrolides ; metabolism ; Point Mutation ; Ribosomal Proteins ; genetics ; Ribosomes ; metabolism ; Saccharopolyspora ; metabolism

OBJECTIVETo observe the effects of close-to-bone needling (CBN) on the expressions of type-Ⅱcollagen, pro-collagen type Ⅱ C-terminal propeptide (PⅡCP) and C-telopeptide of type Ⅱ collagen (CTX-Ⅱ) in rabbits with knee osteoarthritis (KOA).

METHODSAmong 40 New Zealand rabbits, 10 rabbits were selected into a normal group; the remaining 30 rabbits were made into KOA model, and X-ray was used to evaluate the results of model establishment. After the model was successfully made, the rabbits were randomly divided into a model group, a CBN group and a regular acupuncture group, ten rabbits in each one.Rabbits in the CBN group and the regular acupuncture group were treated at "Neixiyan" (EX-LE 4), "Dubi" (ST 35), "Yinlingquan" (SP 9), "Zusanli" (ST 36) and "Liangqiu" (ST 34). The CBN group applied CBN, and the depth of needling was appropriate with needles reaching bone; the regular acupuncture group applied regular acupuncture. The electroacupuncture(EA) device was used in the two groups, 20 min per treatment, once a day.Five days of treatment were taken as one course, and totally 4 courses were given with an interval of 2 days between courses. The normal group received identical fixation as model group. After treatment, magnetic resonance imaging (MRI) was used to perform imaging observation on knee; transmission electron microscopy (TEM) was used to observe the cell structure of knee joint cartilage;HE staining was used to observe the pathological change of knee; TUNEL was used to observe the apoptotic index; the expressions of type-Ⅱ collagen proteins and mRNA were measured by Western-blot and reverse transcription-polymerase chain reaction (RT-PCR); the serum PⅡCP and CTX-Ⅱ levels were measured using enzyme-linked immunosorbent assay (ELISA).

RESULTSAfter treatment, compared with the model group, in the CBN group and regular acupuncture group the articular cavity effusion was reduced without the subchondral bone edema; the cell structure of knee joint cartilage was regular with less apoptosis; the expressions of type-Ⅱ collagen proteins and mRNA were significantly increased (all<0.05), the PⅡCP levels were significantly increased (both<0.05), but the CTX-Ⅱ levels were significantly decreased (both<0.05).The differences of the expressions of type-Ⅱ collagen proteins and mRNA, the levels of PⅡCP and CTX-Ⅱ between the CBN group and the regular acupuncture group were significant (all<0.05); the differences between the CBN group and the normal group were non-significant (all>0.05).

CONCLUSIONSCBN can significantly improve the pathological status of cartilage of KOA, reduce apoptosis, and is likely to regulate the expressions of PⅡCP and CTX-Ⅱ to promote the type-Ⅱ collagen, which is superior to regular acupuncture.

Objective To observe the effects of deep electroacupuncture on carlilage tissue in knee osteoarthritis (KOA) rabbits. Meth-ods 40 New Zealand rabbits were randomly divided into normal group (A, n=10) and model group (n=30). The model group was modeled KOA with Hulth-Telhag way, and identified with X-ray. Then they were divided into no-treated group (B, n=10), deep electroacupuncture group (C, n=10) and routine electroacupuncture group (D, n=10) randomly. The groups C and D accepted electroacupuncture since 6 weeks after modeling, for 4 weeks. They were measured with pH of joint fluid, observed structure and pathology of cartilage under transmission electron microscope, detected apoptosis index, and determined the expression of acid-sensing ion channel 1 (ASIC1), p38 mitogen-activated protein kinases (p38MAPK) and p53 with Western blotting, and distribution of ASIC1 with immunohistochemistry in cartilage tissue. Re-sults The pHs of joint fluid from high to low were ranged as the groups A=C>D>B (P<0.01). The cartilage structure was more complete in the groups A, C and D than in the group B. The apoptosis rates from less to more were ranged as the groups A=C

Background and objectiveSmall cell lung cancer combined with squamous cell carcinoma are rare. hTe aim of this study was to analyze the clinicopathological characteristics and treatment, and explored the prognostic factors of this disease.MethodsBetween January 2004 and December 2012, 58 patients with cytopathologically conifrmed small cell lung cancers combined with squamous cell carcinoma were retrospectively analyzed.Kaplan-Meier methods were used to calculate the survival rate, andLog-rank test was used to examine differences between arms. hTeCox regression model was used to analyze the independent factors affecting the overall survival (OS).Results hTe OS of the 58 patients was 22.7 months with a range of 0.3 to 124.3 months. In univariate analysis, Karnofsky performance score before treatment, extensive disease, tumor stage were the considered prognostic factors affecting the OS rate (P<0.05).Cox multivariate analysis showed that only the tumor-node-metastasis (TNM) stage was the independent prognostic factor (P=0.019). hTe majority of the patients received multimodality therapy and chemotherapy was the main treatment. Distant metastasis was the main reasonfor the treatment failure.ConclusionCombined therapy with chemotherapy as the main treatment should be adopted in therapeutic regimen of the patients with small cell lung cancers combined with squamous cell carcinoma. TNM stage was the independent prognos-tic factor inlfuencing the OS.

Objective To observe the effects of Jinghua Weikang Capsule on NF-κB p65 expression in gastric mucosa of Helicobacter pylori (Hp) infected mice;To discuss its mechanism of action. Methods Totally 44 SPF KM mice were randomly divided into blank group, model group, triple therapy group and Jinghua Weikang group. Except for the blank group, Hp infected model was set up by short term immunosuppression combined with peroral infection. The treatment strategies were triple therapy group with lansoprazole, metronidazole and clarithromycin for 1 week, Jinghua Weikang group with Jinghua Weikang volatile oil for 4 weeks, model group with sterile water for 4 weeks, and blank group free of any treatments. All the subjects were sacrificed in the 4th week after the treatment. NF-κB p65 was detected through real-time PCR and immunohistochemical method. Results The relative quantitative expressions of NF-κB p65 mRNA were 3.27±0.53 in blank group, 2.18±0.50 in control group, 4.19±1.21 in triple therapy group, and 7.70±1.41 in Jinghua Weikang group (χ2=86.303, P<0.001). Compared with model group, the expressions of NF-κB p65 mRNA of other three groups increased statistically differently (P=0.004), and there was also statistical difference between triple therapy and Jinghua Weikang group. Through immunohistochemical tests, the values of NF-κB p65 mean optical density of nucleus were 25.18± 7.42 in blank group, 31.17±2.98 in model group, 30.06±4.69 in triple therapy group, and 26.94±5.19 in Jinghua Weikang group (χ2=10.960, P=0.012). The values of NF-κB p65 mean optical density of cytoplasm had no statistical difference (F=1.922, P=0.134), and the nucleus/cytoplasm values were 0.23±0.06 in blank group, 0.28±0.03 in model group, 0.26±0.04 in triple therapy group, and 0.24±0.04 in Jinghua Weikang group (χ2=14.527, P=0.002). The statistical analysis of the nucleus/cytoplasm values showed significance between blank and model group (P=0.002), while triple therapy group was still equal to model group (P=0.420). Jinghua Weikang group was lower than model group (P=0.022) and equal to blank group (P=0.750). Conclusion Jinghua Weikang Capsule can effectively reduce the expression of NF-κB p65 in protein level in Hp infected mice.