Advances in bacterial adsorption and transport of aromatic compounds / 生物工程学报

Aromatic compounds are a class of organic compounds with benzene ring(s). Aromatic compounds are hardly decomposed due to its stable structure and can be accumulated in the food cycle, posing a great threat to the ecological environment and human health. Bacteria have a strong catabolic ability to degrade various refractory organic contaminants (e.g., polycyclic aromatic hydrocarbons, PAHs). The adsorption and transportation are prerequisites for the catabolism of aromatic compounds by bacteria. While remarkable progress has been made in understanding the metabolism of aromatic compounds in bacterial degraders, the systems responsible for the uptake and transport of aromatic compounds are poorly understood. Here we summarize the effect of cell-surface hydrophobicity, biofilm formation, and bacterial chemotaxis on the bacterial adsorption of aromatic compounds. Besides, the effects of outer membrane transport systems (such as FadL family, TonB-dependent receptors, and OmpW family), and inner membrane transport systems (such as major facilitator superfamily (MFS) transporter and ATP-binding cassette (ABC) transporter) involved in the membrane transport of these compounds are summarized. Moreover, the mechanism of transmembrane transport is also discussed. This review may serve as a reference for the prevention and remediation of aromatic pollutants.

Clinical application of minimal invasive liposuction system in thigh areas of liposuction / 中华医学美学美容杂志

Objective:To investigate the clinical application and surgical effect of minimal invasive liposuction system (MILS) method in the thighs liposuction.Methods:From July 2019 to March 2021, 868 patients were selected for thigh liposuction. Local anesthesia and intravenous anesthesia were used in all operations. The preoperative design and intraoperative procedures were carried out according to the requirements of MILS ? method. There was an average of 2 154.4 ml fat aspirated, and the average operation time was 156.8 min. Results:After 2-18 months of follow-up, 83.3% of the patients were discharged on the day of operation. The thigh line curve of the patients was fluent and the perimeter of upper thigh was significantly reduced. The satisfactory rate was 92.3%, and the incidence of complications was 2.8%. No severe complication occurred.Conclusions:MILS method is a safe and effective procedure to partition thighs liposuction area scientifically and finely. The MILS method makes the lumbar abdominal liposuction operation standardized, reduces unnecessary operation, shortens the liposuction learning curve, reduces the occurrence of complications, and achieves the best aesthetic effect.

Distribution of mesencephalic astrocyte derived neurotrophic factor in human, mouse and rat brain / 解剖学报

Objective To observe the distribution of mesencephalic astrocyte derived neurotrophic factor (MANF) within the brain in human, mice and rats and the comparison across the species. Methods An immunohistochemical method was applied to investigate the expression of MANF and to compare the differences among species in the brain of human specimens ( n = 5) , mice ( n = 6) , and rats ( n = 6). Results The expression of MANF varied in different brain regions in human, mice and rats. In human, in the cortex, the expression of MANF was the highest among all brain regions. In the subcortical areas or the nucleus, brainstem and cerebellum expression level of MANF were relatively lower. The distribution of MANF in mouse and rat brain were slightly different from that in human brain, but generally consistent. Conclusion The distribution pattern of MANF is similar across the species, however, within a species, MANF expression levels varies in different brain regions.

Overexpressing of chalcone isomerase (CHI) gene enhances flavonoid accumulation in Glycyrrhiza uralensis hairy roots / 药学学报

Chalcone isomerase (CHI) is the second rate-limiting enzyme involved in the biosynthetic pathway of flavonoids in Glycyrrhiza uralensis. Based on our previous studies, we selected the specific CHI haplotype (GenBank Accession No. KY115232) to maximize flavonoid accumulation. We constructed a plant binary expression vector for overexpression of this CHI gene by the gene fusion method and transfected the plasmid into Agrobacterium tumefaciens ACCC10060 by electroporation. The recombinant A. tumefaciens ACCC10060 subsequently was used to infect cotyledons and hypocotyls of G. uralensis to obtain transgenic hairy roots. A qRT-PCR method was used to determine the copy number of CHI and a UPLC method was used to assay the content of four flavonoids in different hairy root lines. The qRT-PCR results showed that the copy number of CHI in hairy roots was 1 or 5. UPLC results showed that the content of total flavonoids, liquiritin, liquiritigenin, and isoliquiritigenin in transgenic hairy root samples was significantly higher than that in wild-type samples. This study demonstrates that overexpression of CHI significantly increases the content of flavonoids in hairy roots of G. uralensis. This work provides a theoretical basis for clarifying the function of CHI. Three transgenic hairy root lines of G. uralensis were isolated which can be used to increase the accumulation of licorice flavonoids in vitro.

The role of licorice DXS knockout and overexpression in glycyrrhizic acid biosynthesis / 药学学报

1-Deoxy-D-xylulose-5-phosphate synthase (DXS) is a rate-limiting enzyme involved in the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for terpenoid precursor biosynthesis. DXS plays an essential role in glycyrrhizic acid (GA) biosynthesis. Based on our previous transcriptome study, there was a negative correlation between DXS expression and GA content. Therefore, we explored the regulatory role of DXS in GA biosynthesis using both gene overexpression and gene knockout in a hairy root culture system. DXS was cloned from Glycyrrhiza glabra L. (GenBank Accession No. MN158121). A plant binary expression vector pCA-DXS was constructed by a gene fusion method. The sgRNA sequence was designed based on the first exon of DXS to construct the gene editing vector pHSE-DXS. Hairy roots overexpressing or knocking out DXS were generated through an Agrobacterium-mediated method with licorice hypocotyls as explants. Wild-type hairy roots and negative control hairy roots containing empty plasmids were also evaluated. UPLC was used to determine the GA content in each licorice hairy root line. Results showed that the content of GA in the hairy root group knocking out DXS was significantly higher than that in the wild-type and negative control groups, while in the hairy root group overexpressing DXS was significantly lower, suggesting that DXS plays a negative role in GA biosynthesis. This study provides a foundation for determining the function of DXS in terpenoid metabolism and for further establishment of a molecular regulatory network of GA biosynthesis.

Ferulate 5-hydroxylase gene (F5H) regulation of glycyrrhizic acid biosynthesis determined by gene overexpression and knockout / 药学学报

Ferulate 5-hydroxylase (F5H) is a key enzyme involved in the phenylpropane metabolism pathway. Based on our previous transcriptome sequencing study, F5H played a negative regulatory role in glycyrrhizic acid (GA) biosynthesis. Therefore, in this study we cloned the F5H gene and investigated its regulatory effect on GA accumulation through gene overexpression and knockout. F5H was cloned from Glycyrrhiza glabra L. (GenBank Accession No. MK882511). A plant binary expression vector pCA-F5H was constructed by inserting F5H into pCAMBIA1305.1 at Spe I and Bgl II sites. The sgRNA sequences were designed based on the first exon of F5H. The CRISPR/Cas9 gene editing vector pHSE-F5H was constructed by inserting F5H sgRNA into pHSE401 at two Bsa Ⅰ sites. PCA-F5H and pHSE-F5H were transfected into Agrobacterium tumefaciens ATCC15834, which was used to induce hairy root overexpressing or knocking out F5H with licorice hypocotyl as explants. At the same time, wild type and negative control hairy roots were also generated. UPLC was used to assay the GA content in different hairy root lines, and results showed that the GA content in hairy root lines knocking out F5H was significantly higher, whereas in hairy root lines overexpressing F5H GA content was lower than that in the wild-type and negative control. In this work, through a reverse genetics strategy, the negative regulatory effect of F5H on GA biosynthesis was confirmed through gene overexpression and knockout. This work will lay a foundation for further elucidation of the molecular regulatory network of GA biosynthesis.

Cloning and Bioinformatic Analysis of ARPI Gene from Glycyrrhiza glabra / 中国实验方剂学杂志

Objective::To clone the complementary deoxyribonucleic acid (cDNA) of auxin/indole acetic acid protein (Aux/IAA) from Glycyrrhiza glabra (GgARPI) and analyze its sequence by bioinformatics. Method::RNA was extracted from fresh root of G. glabra, the cDNA sequence of GgARPI gene was cloned by reverse transcription polymerase chain reaction (RT-PCR), then sequencing and bioinformatic analysis were performed. Result::The GgARPI cDNA sequence with the full length of 686 bp was obtained from G. glabra. The full open reading frame (ORF) was 585 bp, encoding 194 amino acid residues. The bioinformatic analysis showed that the protein coded by GgARPI was a stable hydrophilic protein, with a relative molecular weight of 21.95 kDa and isoelectric point of 6.85.It contained no signal peptides or transmembrane domain. Its secondary structure mainly consisted of random coil. An Aux/IAA superfamily was included in the conversed domain. Homology analysis indicated that it had a close evolutionary relationship with leguminous plants, and a distant evolutionary relationship with monocotyledon, such as Setaria italica. Conclusion::GgARPI cDNA sequence is successfully cloned from G. glabra for the first time, which will lay a foundation for studying the function of GgARPI and explaining the molecular regulatory mechanism of biosynthesis of glycyrrhizic acid in G. glabra.

Molecular Cloning and Sequence Analysis of GgUGE Gene in Glycyrrhiza glabra / 中国实验方剂学杂志

Objective::To clone the cDNA sequence of UDP-glucose 4-epimerase (UGE) in Glycyrrhiza glabra and analyze its sequence, so as to explore the potential relationship between the UGE gene and the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis. Method::The cDNA sequence of UGE was cloned from the root of G. glabra by reverse transcription polymerase chain reaction (RT-PCR), then sequenced and analyzed by bioinformatics software. Results::A GgUGE cDNA sequence with the full length of 1 121 bp was obtained. The open reading fame (ORF) of GgUGE was 1 053 bp, encoding 350 amino acid residues. The GgUGE cDNA sequence was submitted to GenBank, and the accession No. was MK638908. Sequence analysis showed that GgUGE was an unstable hydrophilic protein, its average relative molecular weight was 39.02 kDa, and isoelectric point was 6.13. It contained no signal peptides or transmembrane domains. Its secondary structure mainly constituted of α-helix and had a conversed domain of UDP-glucose 4-epimerase superfamily. The homologoue analysis showed that the cDNA and amino acid sequences of GgUGE had the closest evolutionary relationship to Leguminosae and relatively distant evolutionary relationship to Salicaceae. Conclusion::In this study, GgUGE cDNA sequence is successfully cloned from G. glabra for the very first time, which will provide reference for studying the function of GgUGE and explaining the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis in G. glabra.

Optimization of extraction process of total triterpenoid from root of Rose odorata var.gigantean by orthogonal test and its inhibitory effect on contraction of isolated rat intestinal smooth muscle / 药物评价研究

Objective To optimize the extraction technology of total triterpenoid from root of Rose odorata var.gigantean (TTROG) by orthogonal test combined with the contraction effect of TTROG on the isolated intestinal smooth muscle of rats in vitro.Methods UV spectrophotometric method was used to determine the contents of total triterpenoids in the TTROG extractive at the wavelength of 550 nm by taking ursolic acid as standard substance,and vanillin acetic acid as chromogenic reagent.The extraction rate of total triterpenoids was used as index to evaluate the technology based on single factor test,in which three factors were considered as follows:the concentration of extraction solvent,ratio of material to liquid,extraction time,and their interaction on extraction were studied by orthogonal experimental design.The inhibition effect of different extracts obtained from the optimized extraction process on the contraction of intestinal smooth muscle were recorded by tension transducer to the BL-420 biological experimental multi-channel physiological signal acquisition and processing system.The extraction process of TTROG was evaluated by the combination of biological activity and extraction rate with weighting method.Results The optimal extraction conditions of TTROG were as follows:extraction solvent 80％ ethanol,solid-liquid ratio 1∶10,extraction time for 2 h,three times and extraction temperature of 80 ℃.The optimized extraction rate could reach 42.12 mg/g.TTROG obtained using the optimized method showed significantly contraction effect on rat intestinal smooth muscle with dose effect dependence,and the effect on jejunum was the strongest,and the inhibition rate was 41.96％.Conclusion The optimized extraction technology is stable and effective with high extraction rate.TTROG showed the significant inhibitory function on contraction of isolated rat intestinal smooth muscle.

Anticonvulsant effect of intranasal midazolam on maximal electroshock seizure and metrazol seizure model in rats / 军事医学

Objective To evaluate the anticonvulsant effect of midazolam and diazepam when administered nasally on maximal electroshock seizure and metrazol seizure threshold test models .Methods Rats were randomly divided into 7 groups:model group, low-dose, middle-dose and high-dose of midazolam nasal spray groups , diazepam nasal spray in low-dose, middle-dose and high-dose of midazolam nasal spray groups .After the establishment of the maximal electroshock seizure( MES) and metrazol seizure threshold test models ( MST) in rats, the anticonvulsant effects of different doses of midazolam and the clinically used antiepileptic drug diazepam were evaluated and compared .HE staining was used to observe the histopathological changes in the hippocampus , cortex and amygdala in rats .Results Significant anticonvulsant effects were observed on MES and MST in rats pretreated with different dosages of midazolam .In addition , the anticonvul-sant effects of midazolam were stronger than those of diazepam at the same dosage on MES and MST (P<0.05,P<0.01). Histopathological results showed that both midazolam and diazepam could effectively prevent the seizure -induced brain inju-ries, inhibit the increase of microglial cells and the inflammatory cell infiltration in the hippocampus, cortex and amygdala, and reduce the nucleus pycnosis and neuronophagia .Conclusion Midazolam has significantly anticonvulsant and neuropro-tective effects on different seizure models when administered nasally in rats .

Comparison between laparoscopic and open surgical high ligation of the internal spermatic vein / 中华男科学杂志

<p><b>OBJECTIVE</b>To compare the effects of laparoscope-guided high ligation of the internal spermatic vein and open surgery in the treatment of varicocele.</p><p><b>METHODS</b>We collected the clinical data of 85 cases of varicocele treated by laparoscope-guided high ligation of the internal spermatic vein (the laparoscopic group) and another 47 that underwent open surgery (the open group) , and compared the results of treatment between the 2 groups.</p><p><b>RESULTS</b>The operations were successful in all the cases. In the laparoscopic group, the mean operation time was (32.0 +/- 5.9) minutes, with little bleeding, and the mean postoperative hospital stay was (3.0 +/- 1.6) days, with 3 cases (3.5%) of recurrence during the 3-12 months follow-up. In the open group, the mean operation time was (46.0 +/-7.2) minutes and the mean postoperative hospital stay was (7.0 +/- 3.7) days, with 5 cases (10.6%) of recurrence during the same length of follow-up.</p><p><b>CONCLUSION</b>Laparoscopic method for the treatment of varicocele is obviously superior to open surgery for its advantages of minimal invasiveness, fast recovery, sure effectiveness, few complications and low recurrence, particularly for those with bilateral varicocele who experienced recurrence after open surgery.</p>

Retroperitoneal laparoscopic ureterolithotomy for treatment of 33 Patents with ureteral calculi / 国际外科学杂志

Objective To discuss the efficacy and safety of retroperitoneal laparoscopic ureterolithotomy in treatment of ureteral calculi.Methods All 33 cases with upper or midst ureter calculi were treated by laparoscopic ureterolithotomy,ESWL or URSL ineffective for 26 cases.The diameters of calculi ranged from 0.7 to 2.1 cm.Results All the operations were successful.The operation time was 30～120 min with a mean of 55 min.The intraoperative bleeding volume Was 40 ml (20～80 ml) on average no complication Was observed duing the follow-up of 6～12 months.All the hydronephrosis Was alleviated.Conclusion Laparoscopic ureterolithotomy is suitable for ureteral calculi treated unsuccesfully by open surgery.It is effective,safety and miniinvasive.

Effect of lugu Ganoderma lucidum on low-density lipoprotein oxidation and monocyte adhesion to endothelium / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Lugu Ganoderma Lucidum (LGL) on low-density lipoprotein (LDL) oxidation and monocyte adhesion to endothelium (AdM-E) induced by oxydative LDL and advanced glycosylation endproducts (AGE) by using serum pharmacological technique.</p><p><b>METHODS</b>LDL oxidation was determined by measuring the thiobarbituric acid reactive substances in the supernatants, and AdM-E was determined by measuring myeloperoxidase activity of adherent monocyte.</p><p><b>RESULTS</b>Serum derived from rats 0.5 hrs, 1 hr, 2 hrs, 3 hrs after LGL administering 0.12 g/kg once and 0.5 hrs, 1 hr after LGL administering twice showed no significant effect on LDL oxidation, but the serum from rats 2 hrs, 3 hrs after LGL 0.12 g/kg administering twice or from rats after 10 successive days LGL administering in dose of 0.12 g/kg, 0.24 g/kg and 0.72 g/kg, all could lower the LDL oxidation (P < 0.05). Besides, the serum from rats with 10 days LGL administering of all dosages also could inhibit AdM-E induced by AGE (P < 0.05), and those of 0.24 g/kg and 0.72 g/kg could inhibit AdM-E induced by oxydative LDL (P < 0.05).</p><p><b>CONCLUSION</b>LGL could decrease LDL oxidation and AdM-E induced by AGE or oxydative LDL.</p>

Ureteroscopic Holmium Laser Lithotripsy for Ureteral Calculi after Failure of ESWL:Report of 89 Cases / 中国微创外科杂志

Objective To evaluate the efficacy of ureteroscopic holmium laser lithotripsy for ureteral calculi after failed extracorporeal shock-wave lithotripsy(ESWL).Methods A total of 89 patients with ureteral calculi were treated by ureteroscopic holmium YAG laser lithotripsy after failed ESWL.Among them,69 cases were complicated with ploypi or calculi surrounded by granuloma tissues,which were melted by holmium laser simultaneously;4 cases were complicated with distal ureteral stenosis,and received open surgery for resection of the stenotic segment.Results Of the patients,calculi were fragmented by one operation in 81 cases.The success rate was 91%.67 cases were stone free in one week and 14 cases in two weeks.In 4 cases,calculi were flushed into the pelvis,and were cured by open surgery.Conclusions Ureteroscopic holmium laser lithotripsy is an effective and safe method for ureteral calculi after failed ESWL.It should be used as the first choice for the disease.

Laparoscopic treatment of ureteropelvic junction obstruction / 中国微创外科杂志

Objective To evaluate the clinical efficiency of laparoscopic operation for the treatment of ureteropelvic junction obstruction(UPJO).Methods A total of 19 patients with UPJO underwent laparoscopic dismembered pyeloplasty.The operation was performed under general anesthesia.The patients were maintained in a lateral decubitus on unaffected side.After the establishment of pneumoperitoneum,3~4 trocars were introduced at proper sites.During the operation,the lateral peritoneum was opened and the ureteropelvic junction obstruction was dissociated and dissected.The end-to-end anastomosis of the renal pelvis and ureter was conducted and a double-J stent was placed in the ureter.Results All the operations were performed successfully.The operation time was 110~240 min(mean,150 min),the blood loss was 50~100 ml(mean,80 ml),and the postoperative hospitalization time,6~10 d(mean,7.8 d).No severe complications occurred.Follow-up assessment with intravenous urography for 3~15 months(mean,6 months) in 14 patients showed no ureteral stricture.Conclusions Laparoscopic dismembered pyeloplasty is an effective and safe procedure in the treatment of UPJO.It can replace the open surgery.

Holmium laser percutaneous nephrolithotomy for treating upper ureteral callui: Report of 132 cases / 中国微创外科杂志

Objective To explore the safety and efficiency of holmium laser percutaneous nephrolithotomy in the treatment of upper ureteral calcui.Methods A retrospective analysis was made on 132 cases of upper ureteral calcui treated by holmium laser percutaneous nephrolithotomy.After retrograde placement of the ureteral catheter,a track from the skin surface into the middle renal calyx was established under the guidance of a C-arm fluoroscopy.Then a ureteroscope was passed to introduce a holmium laser lithotriptor.Stones were broken up and a nephrostomy tube was placed in the incision to carry fluid from the kidney.Results Holmium laser percutaneous nephrolithotomy was completed on one session in all the 132 cases(155 sides),including 116 cases of single-tract nephrolithotomy and 16 cases of double-tract nephrolithotomy.The stone-free rate was 95.5%(148/155).The duration of operation was 30~320 min(mean,108 min).The intraoperative blood loss was 50~200 ml(mean,100 ml).No blood transfusion was required and no postoperative hemorrhage happened.No analgesics were needed after operation.The nephrostomy tube was left for 3~8 d(4.6 d).The length of postoperative hospital stay was 5~11 d(mean,7.4 d).Follow-up observations in the 132 cases for 1~6 months(mean,3 months) showed complete relief of symptoms.B-ultrasonography showed a mean of renal pelvis separation of 15 mm,which had decreased by 9~28 mm postoperatively.Conclusions Holmium laser percutaneous nephrolithotomy is safe and effective in the treatment of upper ureteral calcui.