Objective To study the clinical characteristics and iduronate-2-sulfatase (IDS)gene mutation of one child patient with mucopolysaccharidosis type Ⅱ (MPS Ⅱ).Methods All the 9 exons of IDS gene were amplified by polymerase chain reaction (PCR)technlogy.The PCR products were screened by direct gene Sanger sequencing.Results A missense muta-tion (c.445T>C)on exon 4 was found after the analysis of the gene sequencing results of PCR products in this patient’s IDS gene.Thi smutation leaded to the 149th codon TCT encoded serine into a CCT encoding proline (p.Ser149Pro).Mean-while,the IDS gene in the parents were widetpye,so this was a de novo mutation.Conclusion The de novo mutation of IDS gene is the cause of our patient with?mucopolysaccharidosis,one novel mutation (p.Ser149Pro)was identified.

[ Objective ] To investigate the therapeutic effect of Chinese herbal medicines, which have the actions of dispersing stagnated liver-Qi and promoting bile secretion, clearing heat and removing damp, activating blood and removing blood-stasis, in the treatment of the third-level or above intrahepatic biliary duct stone. [Methods] After the routine surgical treatment including cholecystectomy, choledochotomy with T-tube drainage and choledochoscope lithotomy, 42 cases of third-level or above intrahepatic biliary duct stone received Dandao Paishi Mixture (DPM, mainly composed of Herba Lysimachiae, Radix Clematidis, Herba Artemisiae Scopariae, Radix Aucklandiae, Rhizoma Curcumae Longae, Radix Curcumae, etc.) for oral use. The effects of DPM on the intrahepatic biliary duct stone were evaluated after treatment. [Results] Before the treatment of DPM, the stones in third-level or above intrahepatic biliary ducts cannot be taken out; after the treatment of DPM, T-tube reverse biliary contrast examination showed that stones were removed in 39 (92.9%) cases, stones still existed in the common bile duct in 3 cases and then were removed by lithotomy through T-tube sinus of the common bile duct with Olympus fiber choledochoscope 8 weeks later. [Conclusion] DPM has good lytholytic and lithagogue effects for the treatment of third-level or above intrahepatic biliary duct stone and for intrahepatic biliary duct incarcerated stone. It can also reduce the postoperative residue of stone and recurrence rate.

By engineering the subsite +1 of cyclodextrin glycosyltransferase (CGTase) from Paenibacillus macerans, we improved its maltodextrin specificity for 2-O-D-glucopyranosyl-L-ascorbic acid (AA-2G) synthesis. Specifically, we conducted site-saturation mutagenesis on Leu194, Ala230, and His233 in subsite +1 separately and gained 3 mutants L194N (leucine --> asparagine), A230D (alanine --> aspartic acid), and H233E (histidine --> glutamic acid) produced higher AA-2G yield than the wild-type and the other mutant CGTases. Therefore, the 3 mutants L194N, A230D, and H233E were further used to construct the double and triple mutations. Among the 7 obtained combinational mutants, the triple mutant L194N/A230D/H233E produced the highest AA-2G titer of 1.95 g/L, which was increased by 62.5% compared with that produced by the wild-type CGTase. Then, we modeled the reaction kinetics of all the mutants and found a substrate inhibition by high titer of L-AA for the mutants. The optimal temperature, pH, and reaction time of all the mutants were also determined. The structure modeling indicated that the enhanced maltodextrin specificity may be related with the changes of hydrogen bonding interactions between the side chain of residue at the three positions (194, 230 and 233) and the substrate sugars.
Ascorbic Acid ; analogs & derivatives ; chemistry ; Glucosyltransferases ; genetics ; metabolism ; Hydrogen Bonding ; Kinetics ; Mutagenesis, Site-Directed ; Paenibacillus ; enzymology ; Polysaccharides ; chemistry ; Protein Engineering ; Substrate Specificity ; Temperature

Objective To investigate cartilage oligomeric matrix protein( COMP) gene mutation in a three-generation pedigree with two cases of pseudoachondroplasia, and to definitize genotype-phenotype correlation. Methods The clinical data and peripheral blood were collected from the patients with pseudoachondroplasia and their family members. All the 19 exons and their flanking sequences of COMP gene in two patients and three unaffected family numbers and 50 unrelated individuals were analyzed by PCR amplification and direct sequencing. Results The proband, a 6-year-old girl presented with typical clinical features of pseudoachondroplasia, including disproportionate short limb dwarfism, staggering gait, double genu varus deformity, and wider clinical and imaging long bone metaphysis. The 33-year-old father showed a similar manifestation including disproportionate short limb dwarfism and double genu varus deformity, and was performed correcting operation on lower limbs for double genu varus at the age of 10 years. DNA sequencing analysis of the COMP gene revealed a del mutation ( c. 1417 1419delGAC)in exon 13 in two patients with pseudoachondroplasia, but not in the other unaffected members from the pedrgree and 50 control subjects. Conclusion A del mutation c. 1417 1419delGAC of COMP gene may contribute to the disease in the pedigree.

OBJECTVE To study the improvement effects of obacunone on renal interstitial fibrosis （RIF） in unilateral ureteral obstruction （UUO） model mice， and to investigate its mechanism based on ferroptosis mediated by nuclear factor erythroid 2- related factor 2（Nrf2）/glutathione peroxidase 4 （GPx4） signaling pathway. METHODS Thirty mice were randomly divided into sham operation group， model group， irbesartan group （positive control， 20 mg/kg）， obacunone low-dose and high-dose groups （10， 40 mg/kg）， with 6 mice in each group. Except for sham operation group， UUO model was established by ligation of unilateral ureter in other groups. After operation， administration groups were given intraperitoneal injection of relevant medicine， and sham operation group and model group were given intraperitoneal injection of constant volume of normal saline， once a day， for 7 consecutive days. The levels of creatinine （Cr） and blood urea nitrogen （BUN）， the content of malondialdehyde （MDA） and the activities of total superoxide dismutase （T-SOD） in serum and the concentration of Fe2+ in renal tissue were all detected. HE staining and Masson staining were performed to observe the morphology and the fibrosis of renal tissues. Immunohisto- chemical staining was used to determine expressions of the fibronectin （Fn）， α-smooth muscle actin （α-SMA）， GPx4 964083717@qq.com and Nrf2 in renal tissue. Western blot and real-time quantitative polymerase chain reaction （RT-qPCR） were used E-mail：834300205@qq.com to detect the protein and mRNA levels of Fn， α-SMA， Nrf2， GPx4 and SLC7A11 in the renal tissues. RESULTS Compared with sham operation group， serum levels of Cr and BUN， the concentration of Fe2+ in renal tissue， the protein and mRNA levels of Fn and α-SMA in model group were increased significantly （P＜0.05）， while the activity of T-SOD in serum， protein and mRNA expressions of Nrf2， GPx4， SLC7A11 in kidney tissue were significantly decreased （P＜0.05）； in the kidney tissue， the renal tubules were dilated， the collagen deposition was obvious， the fibrous bands were thicker and darker， and the renal interstitial inflammatory cells infiltrated significantly. After intervened with obacunone， the levels of above indexes （except for mRNA expression of SLC7A11 in obacunone low-dose group） in serum and renal tissue were reversed significantly （P＜0.05）， and pathological damage and collagen deposition of kidney tissue were alleviated. CONCLUSIONS Obacunone can improve renal interstitial fibrosis of UUO model mice， the mechanism of which may be associated with activating the Nrf2/GPx4 pathway and then inhibiting ferroptosis to relieve RIF in UUO model mice.

By reviewing ancient materia medica， medical books， prescription books and modern literature， this paper conducted a systematic research on name， origin， medicinal parts， producing area， quality， harvesting and processing methods， functions and toxicity of Polygoni Multiflori Radix（PMR） and Polygoni Multiflori Caulis（PMC） in famous classical formulas. It was found that the name of Heshouwu was first found in the Biography of Heshouwu and originated from its discoverer， and then still in use today. Since the Song dynasty， Heshouwu has been included in the materia medica with Fallopia multiflora as the mainstream origin. Since the Ming dynasty， in addition to F. multiflora， Cynanchum bungei， C. wilfordii， C. auriculatum have been used as the origin of Heshouwu. Heshouwu is widely distributed， the production areas recorded in ancient times are mainly Luchuan， Guangxi and Ganzhou， Jiangxi， and in modern times， Deqing， Guangdong and other places are respected as the geo-authentic habitats. Its origin processing is mostly dug out， washed， sliced and dried in the sun. Modern literature concludes that the quality of PMR is better in terms of weight， solid quality and powder， while PMC is better in terms of uniformity and purplish-red skin. In the Qing dynasty and before that， it was recorded in the materia medica that PMR was harvested mainly in late spring， mid-summer and early autumn， in modern times， it was harvested mainly in spring and autumn， while PMC was harvested in autumn. In Song dynasty， the processing method of PMR was mainly soaked with rice-washed water， the Ming dynasty for black bean steaming method， the Qing dynasty for steaming followed by black bean juice maceration， while in modern times， it is mostly sliced and steamed with black bean juice and yellow rice wine until the juice is absorbed. PMC is prepared by removing impurities， washing or moistening， and cutting into sections to dry. During the five dynasties， PMR was used to treat infertility as well as gastrointestinal diseases and blood in the stool caused by wind chill， and during the Song dynasty， it was mostly used raw products for the treatment of scrofula and carbuncles， and in the Yuan and Ming dynasties， raw and processed products were used， and Polygoni Multiflori Radix Praeparata was used to treat hemorrhoids， waist and knee soreness. PMC is mainly used for treating insomnia， fatigue and sweating， wind sores and scabies， etc. In the Song dynasty， the ancients considered that PMR was non-toxic， but its toxicity was gradually discovered after the Ming dynasty， and the toxicity mechanism has not been clearly analyzed yet. Based on the results of the textual research， it is recommended that F. multiflora be used as the base for development of famous classical formulas containing PMR， and from the safety perspective， it is recommended to use raw products with caution， oral administration must be used processed products. PMC is recommended to use raw products.