Incidence and risk factors of anastomotic leak after transanal total mesorectal excision in China: a retrospective analysis based on national database / 中华胃肠外科杂志

Objective: Transanal total mesorectal excision (taTME) was a very hot topic in the first few years since its appearance, but now more introspections and controversies on this procedure have emerged. One of the reasons why the Norwegian Ministry of Health stopped taTME was the high incidence of postoperative anastomotic leak. In current study, the incidence and risk factors of anastomotic leak after taTME were analyzed based on the data registered in the Chinese taTME Registry Collaborative (CTRC). Methods: A case-control study was carried out. Between November 15, 2017 and December 31, 2020, clinical data of 1668 patients undergoing taTME procedure registered in the CTRC database from 43 domestic centers were collected retrospectively. After excluding 98 cases without anastomosis and 109 cases without complete postoperative complication data, 1461 patients were finally enrolled for analysis. There were 1036 males (70.9%) and 425 females (29.1%) with mean age of (58.2±15.6) years and mean body mass index of (23.6±3.8) kg/m(2). Anastomotic leak was diagnosed and classified according to the International Study Group of Rectal Cancer (ISREC) criteria. The risk factors associated with postoperative anastomotic leak cases were analyzed. The impact of the cumulative number of taTME surgeries in a single center on the incidence of anastomotic leak was evaluated. As for those centers with the number of taTME surgery ≥ 40 cases, incidence of anastomic leak between 20 cases of taTME surgery in the early and later phases was compared. Results: Of 1461 patients undergoing taTME, 103(7.0%) developed anastomotic leak, including 71 (68.9%) males and 32 (31.1%) females with mean age of (59.0±13.9) years and mean body mass index of (24.5±5.7) kg/m(2). The mean distance between anastomosis site and anal verge was (2.6±1.4) cm. Thirty-nine cases (37.9%) were classified as ISREC grade A, 30 cases (29.1%) as grade B and 34 cases (33.0%) as grade C. Anastomotic leak occurred in 89 cases (7.0%,89/1263) in the laparoscopic taTME group and 14 cases (7.1%, 14/198) in the pure taTME group. Multivariate analysis showed that hand-sewn anastomosis (P=0.004) and the absence of defunctioning stoma (P=0.013) were independently associated with anastomotic leak after taTME. In the 16 centers (37.2%) which performed ≥ 30 taTME surgeries with cumulative number of 1317 taTME surgeries, 86 cases developed anastomotic leak (6.5%, 86/1317). And in the 27 centers which performed less than 30 taTME surgeries with cumulative number of 144 taTME surgeries, 17 cases developed anastomotic leak (11.8%, 17/144). There was significant difference between two kinds of center (χ(2)=5.513, P=0.019). Thirteen centers performed ≥ 40 taTME surgeries. In the early phase (the first 20 cases in each center), 29 cases (11.2%, 29/260) developed anastomotic leak, and in the later phase, 12 cases (4.6%, 12/260) developed anastomotic leak. The difference between the early phase and the later phase was statistically significant (χ(2)=7.652, P=0.006). Conclusion: The incidence of anastomotic leak after taTME may be reduced by using stapler and defunctioning stoma, or by accumulating experience.

Role and mechanism of splenic myeloid-derived suppressor cells in sepsis-induced adrenal injury in mice / 中华危重病急救医学

Objective:To investigate the role and mechanism of splenic myeloid-derived suppressor cells (MDSCs) in sepsis-induced adrenal injury (SAI).Methods:Thirty male C57 mice aged 6-8 weeks were randomly divided into normal control group ( n = 5), sham operation group (Sham group, n = 5), sepsis model group [cecal ligation and perforation (CLP) group, n = 10] and sepsis+splenectomy group (CLPS group, n = 10). The sepsis model of mice was reproduced by CLP method. In Sham group, only the cecum was opened and separated, then closed, without CLP. In CLPS group, the spleen was removed before CLP. In normal control group, no challenge was given. After 24 hours, the rats were sacrificed by anesthesia, and peripheral blood, spleen, bone marrow, and bilateral adrenal glands were harvested. The pathological of adrenal gland was assessed by hematoxylin-eosin (HE) staining under optical microscope. The ratio of MDSCs in peripheral blood, spleen and bone marrow was determined by flow cytometry. The expressions of MDSCs surface antigen CD11b, Gr-1 and interleukins (IL-6, IL-1β) mRNA in adrenal tissue were measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Western Blot was used to detect the expressions of mammalian rapamycin target protein (mTOR) pathway related proteins including total mTOR (T-mTOR), phosphorylation of mTOR (p-mTOR) and caspase-3. Results:The adrenal cortex and medulla of the normal control group and Sham group were intact and the structure was clear under optical microscope, while in the CLP group, the adrenal gland showed edema, cortical hemorrhage and cell edema. Compared with the CLP group, the adrenal tissue injury was significantly reduced in the CLPS group. Compared with the normal control group and Sham group, MDSCs ratio in the peripheral blood was significantly increased and significantly reduced in the spleen in the CLP group, but there was no significant difference in bone marrow, the expression levels of CD11b, Gr-1, IL-6, IL-1β mRNA and caspase-3 protein were increased significantly and p-mTOR protein expression was significantly decreased in adrenal tissue, there was no significant difference in the expression of T-mTOR protein. Compared with the CLP group, in the CLPS group, the MDSCs ratio in the peripheral blood was significantly decreased (0.143±0.011 vs. 0.324±0.023, P < 0.01), the expression levels of CD11b, Gr-1, IL-6 , IL-1β mRNA and caspase-3 protein in adrenal gland were significantly decreased [CD11b mRNA (2 -ΔΔCt): 2.90±0.56 vs. 5.74±0.13, Gr-1 mRNA (2 -ΔΔCt): 2.71±0.14 vs. 4.59±0.46, IL-6 mRNA (2 -ΔΔCt): 2.44±0.64 vs. 5.17±1.04, IL-1β mRNA (2 -ΔΔCt): 3.58±0.52 vs. 4.44±0.26, caspase-3 protein (caspase-3/GAPDH): 0.05±0.01 vs. 0.13±0.02, all P < 0.01], the p-mTOR protein expression was significantly increased (p-mTOR/GAPDH: 0.61±0.11 vs. 0.27±0.04, P < 0.01). Conclusions:The spleen is the major source of MDSCs in SAI. Splenectomy can attenuate SAI by reducing mobilization of MDSCs and activating the mTOR signaling pathway.

Expression characteristics of Piezo1 protein in stress models of human degenerative chondrocytes / 中国骨伤

<p><b>OBJECTIVE</b>To explore the expression characteristics of new mechanosensitive ion channel Piezo1 protein in stress models of human degenerative chondrocytes.</p><p><b>METHODS</b>The stress stimulation model of human degenerative chondrocytes in vitro was constructed. Multi-channel cell stretch stress loading system FX-4000T was used to treat chondrocytes. According to the results of pre-test, the loading frequency of 0.5 Hz and the cell elongation of 20% were loaded. According to cell processing time, it was divided into 0 h, 2 h, 12 h, 24 h and 48 h mechanical stress group. The RT-PCR and Western-blot were used to test the expression of the Piezo1, also the Laser scanning confocal microscope (LSCM) was used to test the intensity of the fluorescence of the Piezo1.</p><p><b>RESULTS</b>(1)The result of the RT-PCR showed that the expression of the Piezo1 in the 2 h group was higher than the 0 h group(13.917, 0.037 1, <0.05). The expression of the piezo1 in the 24 h group was the highest. While the expression of the piezo1 in the 48 h group was lower than the expression of the piezo1 in the 24 h group(13.917, 0.049 5, <0.05). (2)The result of the Western-blot showed that the 2 h group was higher than the 0 h group(19.341, 0.037 1, <0.05). The expression of the 24 h had the highest expression which was higher than the 48 h group(19.341, 0.017 7, <0.05). (3)The Piezo1 protein was extensively expressed in the cytoplasm and nucleus of the nucleus pulposus cells. And with the increase of stress processing time, the fluorescence intensity of the protein also increased.</p><p><b>CONCLUSIONS</b>In human degeneration cartilage cells, the new mechanio sensitive ion channel Piezo1 protein has a trace expression. After loading periodic mechanical tensile force, the expression of Piezo1 protein increases with time dependence.</p>

Expression of CLAUDIN-11 in the testicular tissue of the patient with non-obstructive azoospermia and its clinical significance / 中华男科学杂志

<p><b>Objective</b>To study the expression of CLAUDIN-11 in the testis tissue of non-obstructive azoospermia (NOA) patients with different severities and investigate its clinical significance.</p><p><b>METHODS</b>Sixty-two NOA patients were divided into a hypospermatogenesis (HS) group (n = 30) and a Sertoli cell only syndrome (SCO) group (n =32). The expression of CLAUDIN-11 in the testicular tissue of the patients was detected by immunohistochemistry, that of CLAUDIN-11 mRNA determined by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the levels of serum reproductive hormones measured by chemiluminescent immunoassay.</p><p><b>RESULTS</b>Immunohistochemistry showed that the expression of CLAUDIN-11 was mainly in the cytoplasm of the Sertoli cells around the seminiferous tubule wall in the HS group, but diffusely distributed in the membrane of the Sertoli cells in the SCO group. RT-qPCR revealed a significantly lower expression of CLAUDIN-11 mRNA in the HS than in the SCO group (0.008 ± 0.001 vs 0.013 ± 0.002, t = 10.616, P<0.01). The level of serum luteotropic hormone (LH) was also markedly lower in the HS than in the SCO group (［3.62 ± 1.34］ vs ［4.96 ± 3.10］ IU/L, P<0.05) and so was that of follicle-stimulating hormone (FSH) (［5.36 ± 2.80］ vs ［10.65 ± 9.18］ IU/L, P<0.05).</p><p><b>CONCLUSIONS</b>The up-regulated expression of CLAUDIN-11 in Sertoli cells may play an important role in the development and progression of spermatogenic dysfunction in NOA patients.</p>

The clinical effect of acupoint application for the patients with the chronic insomnia with liver-stagnation and Spleen-deficiency syndrome / 国际中医中药杂志

Objective To observe the clinical effect on sleep improvement about acupoint application in treating the chronic insomnia of Liver-stagnation and Spleen-deficiency Syndrome.Methods A total of 68 patients with chronic insomnia with Liver-stagnation and Spleen-deficiency Syndrome were randomly and blindly divided into treatment group and control group by registration order. In the treatment process, 1 was eliminated and 33 were completed in the treatment group, 2 were eliminated and 32 were completed in the control group. The treatment group underwentAn Mian Tietherapy half an hour before bed and the control group underwent placebo therapy in the same way of treatment group. Two groups were treated for 40 days and followed-up visit six months. The change of Pittsburgh Sleep Quality (PSQI), Index Insomnia Severity Index (ISI) and TCM Syndromes Scale were detected.Results The clinical total effective rate of treatment group was 72.7% and the control group was 9.4%, and the difference was statistically significant (χ2=46.977,P<0.01). The PSQI scores after treatment (7.55 ± 1.52vs. 13.90 ± 2.44,t=148.165), and follow up 1 month (8.97 ± 2.51vs. 13.17 ± 2.79,t=37.926) in the treatment group were significantly lower than those in the control group (P<0.01). The ISI scores after treatment (7.03 ± 3.37vs. 20.89 ± 4.40,t=73.75), and follow up 1 month (9.81 ± 3.16vs. 19.41 ± 3.66,t=40.79) in the observation group were significantly lower than those in the control group (P<0.01). The TCM Syndromes Scale scores after treatment (2.05 ± 1.09vs. 6.98 ± 1.23,t=17.116), and follow up 1 month (4.06 ± 1.59vs. 6.83 ± 0.91,t=68.055) and follow up 6 month (5.12 ± 1.84vs. 7.19 ± 1.07,t=27.716) in the observation group were significantly lower than those in the control group (P<0.01).Conclusions Acupoint application could obviously change the sleep quality and Chinese medicine symptom in chronic insomnia of Liver-stagnation and Spleen-deficiency Syndrome.

Determination of N,N-Dimethylaniline in Quetiapine Fumarate by LC-MS / 医药导报

Objective To establish a LC-MS analytical method for determination of N,N-dimethylaniline which is genotoxic impurity in quetiapine fumarate.Methods The method was achieved by an Waters ACQUITY UPLC CSHTM PhenylHexyl(2.1 mm× 100 mm,1.7 μm) utilizing a mobile phase of buffer-methanol(900∶ 100) (A)-acetonitrile(B) with gradient elution at the flow rate of 0.4 mL·min-1.The temperature of column was set at 50 ℃;The DIONEX Ultimate 3000 HPLC-AB Science 4000 QTrap Tripling Four bar LC-MS to detect N,N-Dimethylaniline (ESI source,in MRM positive mode).Results Standard curve was linear in the range of 0.4-8.0 ng(r =0.999 3);The limit of detection was 0.2 ng;The limit of quantification of N,N-dimethylaniline was 0.4 ng,respectively.The average recovery of N,N-dimethylaniline was 103.3 ％;RSD was 4.3％ (n =9),respectively.Conclusion The method is convenient and sensitive for the determination of N,N-dimethylaniline in quetiapine fumarate.

Diversity and distribution of the threatened medicinal vascular plants in Lancang / 中国中药杂志

The rich diversity in medicinal plants provides an important material basic for the development of Traditional Chinese medicine in China. It is important to explore the present situation of medicinal plants within special regions in order to provide scientific instructions for their sustainable protection and exploitation and utilization. In this study, we carried out the field survey according to the guideline of national survey of Chinese material medica resources and the guideline of plant species diversity survey and estimation at county level with the line transect method. With the field surveyed data, we explored the diversity and distribution of the threatened medicinal vascular plants in Lancang. We found that there were 33 species of the threatened medicinal vascular plants in this county. These species were from 23 genera and 17 families, and were composed of one critical endangered, 10 endangered and 22 vulnerable species. They were widely distributed across the whole county and were most concentrated in the town of Nuozhadu, Fazhanhe, Nuofu and Zhutang, which were located in the southeastern, southwestern and western of Lancang, respectively. We also found that the plant species richness followed a unimodal pattern along elevation. In addition, we found that the areas of Nuozhadu Nature Reserve in Lancang only covered six threatened medicinal vascular plants, while most of the regions with high species richness were not well protected. Therefore, we proposed to make more efforts to improve the protection measurements in order to better protect and utilize the medicinal plants in Lancang.

A Molecular-Imprinted Sensor for Trace Detection of Gibberellin Based on Ferrocenecarboxylic Acid Multiply Marked Dendrimer / 分析化学

A novel strategy employing dendrimer to multi-label the template molecule to improve the sensitivity of molecularly imprinted electrochemical sensor was proposed. The determination relies on a competition reaction between poly ( diethylene-triaminepenta-acetic acid )-glycol ester ( PDTPA )-ferrocene-carboxylic acid ( FcA ) labeled gibberellins acid 3 ( GA3 ) and GA3 in the sample. Since one cavity corresponds with multiple FcA, instead of only one FcA, the intensity of the detecting signal was greatly enhanced, so was the sensitivity of the sensor. Experimental results showed that the molecularly imprinted electrochemical sensor was sensitive to GA3 detection at a concentration from 2. 0í10-9 to 1. 0í10-7 mol/L, with a detection limit of 9. 3í10-10 mol/L. In addition, the sensor had good reproducibility and its feasibility was verified in the analysis of series of real beer samples.

Computational Analysis of miRNA and Target mRNA Interactions: Combined Effects of The Quantity and Quality of Their Binding Sites / 生物化学与生物物理进展

MicroRNAs (miRNAs) act by binding to complementary sites on target messenger RNA (mRNA) to induce mRNA degradation and/or translational repression. To investigate the influence of miRNAs at transcript levels, two human miRNAs (miR-1 and miR-124) were transfected into HeLa cells and microarrays used to examine changes in the mRNA profile showed that many genes were downregulated and that the fold decreases in levels of these target mRNAs differed remarkably. Features depicting interactions between miRNAs and their respective target mRNAs, such as the number of putative binding sites, the strength of complementary matches and the degree of stabilization of the binding duplex, were extracted and analyzed. It was found that, for a given target mRNA, both the quality and quantity of miRNA binding sites significantly affected its degree of destabilization. To delineate these types of interactions, a simple statistical model was proposed, which considers the combined effects of both the quality and quantity of miRNA binding sites on the degradation levels of target mRNAs. The analysis provides insights into how any animal miRNA might interact with its target mRNA. It will help us in designing more accurate methods for predicting miRNA targets and should improve understanding of the origins of miRNAs.

The study of SVM-based recognition of particles in urine sediment / 中国医疗器械杂志

This article used support vector machine (SVM) algorithm to recognize the particles in urine sediment in this paper. After feature extraction, cross-validation method and the contour chart of the accuracy were implemented to select the kernel function and the parameters of SVM, and according to the characteristics of SVM classifier and sample data, Multi-SVMs with two-level-classifier was successfully designed and A classification matrix was eventually obtained. The evaluation by using clinical data and comparative results with the artificial neural network have demonstrated that the proposed algorithm gets better results.

Computational Analysis of miRNA and Target mRNA Interactions:Combined Effects of The Quantity and Quality of Their Binding Sites / 生物化学与生物物理进展

MicroRNAs(miRNAs) act by binding to complementary sites on target messenger RNA(mRNA) to induce mRNA degradation and/or translational repression.To investigate the influence of miRNAs at transcript levels,two human miRNAs(miR-1 and miR-124) were transfected into HeLa cells and microarrays used to examine changes in the mRNA profile showed that many genes were downregulated and that the fold decreases in levels of these target mRNAs differed remarkably.Features depicting interactions between miRNAs and their respective target mRNAs,such as the number of putative binding sites,the strength of complementary matches and the degree of stabilization of the binding duplex,were extracted and analyzed.It was found that,for a given target mRNA,both the quality and quantity of miRNA binding sites significantly affected its degree of destabilization.To delineate these types of interactions,a simple statistical model was proposed,which considers the combined effects of both the quality and quantity of miRNA binding sites on the degradation levels of target mRNAs.The analysis provides insights into how any animal miRNA might interact with its target mRNA.It will help us in designing more accurate methods for predicting miRNA targets and should improve understanding of the origins of miRNAs.