Adolescent ; Adult ; Cholecystitis ; complications ; drug therapy ; surgery ; Cholecystolithiasis ; complications ; drug therapy ; surgery ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; drug therapy ; etiology ; surgery ; Phytotherapy ; Postoperative Period ; Sphincterotomy, Endoscopic

PNS (Panax notoginseng saponins) is the main medical bioactive component in Panax notoginseng. The medical value of PNS cannot be extended because of its low production. With the deep study of saponins biosynthetic pathway, the control of PNS biosynthesis through metabolic engineering has gradually become possible. In this study, the Squalene synthase (SS) over-expression vector was established. By the way of agrobacterium-mediated method, the vector was transfered and integrated into the Panax notoginseng genome. The result of the PCR detection and the saponin content detection shows that over-expression SS is able to produce high level of Panax notoginseng saponins, and confirms the regulatory function of SS in the biosynthesis of ginsenosides in Panax notoginseng. It provides a theoretical basis and technical basis for the construction of PNS homologous or heterologous efficient expression system in the future.

Objective To investigate the clinical characteristics and mutations of guanosine triphosphate eyclohydrolase (GCH) Ⅰ gene in patients with dopa-responsive dystonia (DRD). Methods Five families with 18 affected family members and 17 patients with sporadic DRD were examined. Patients were allocated into 3 groups according to onset time, either in childhood, or in adolescence or adult. Interview, physical examination, psychologic testings and CT/MR scan were performed. Mutation screening was performed on 26 patients and 1 normal family nember. Thirty-five healthy control subjects were matched for age and sex. Statistical analysis were conducted with the use of SPSS 13.0 computer software. Results(1)Most of patients started with dystonia. The main clinical manifestation was dystonia too. There was no difference among 3 groups.(2) There were significant differences in diurnal fluctuation among 3 groups(15/15,6/6,7/14, χ2=13.125,P=0.001). Diurnal fluctuation negatively correlated with age (r=-0.720, P<0.01).(3)The differences in postural tremor were also found among 3 groups (7/15,5/6,1/14, χ2=8.073, P=0.018). Postural tremor positively correlated with age (r=0.399, P=0.018).(4)There were differences in exaggeration of tendon among three groups(11/15,1/6,4/14, χ2=8.309, P=0.016). Exaggeration of tendon reflexes negatively correlated with age (r=-0.429, P=0.010).(5)The scores of Hamilton Depression Scale and Hamilton Anxiety Scale in patients were higher than those in controls.(6)DNA sequencing revealed a heterozygous A224G missense mutation(Tyr75Cys)located within exon 1 in one autosomal dominant inheritance family. Conclusions The manifestations of DRD varies. The clinical course is closely correlated with age. A missense mutation(A224G)in coding region of the GCH 1 gene probablyleads to the occurrence of DRD.

Objective To evaluate the therapeutic effect of acupoint catgut embedding on gastrointestinal dysfunction after surgery in patients with severe traumatic brain injury.Methods Sixty-eight consecutively hospitalized patients with gastrointestinal dysfunction after surgery in patients with severe traumatic brain injury admitted to the Department of Intensive Care Unit (ICU) in the First Affiliated Hospital of Zhejiang University of Chinese Medicine from January 2015 to December 2015 were enrolled.Six of those patients were withdrawn from the course of observation,and actually only 62 patients entered the study.They were divided into an observation group (32 cases) and a control group (30 cases) by random number table.All the patients of two groups were given conventional treatment of western medicine for consecutive 7 days.The observation group was additionally treated with acupoint catgut embedding.The control group took mosapride tablets via a nasogastric tube 5 mg,3 times a day.Intra-abdominal pressure (IAP),gastric residual volume (GRV),time for reaching standard enteral nutrition,serum diamine oxidase(DAO),D-Lactate activity,incidence of hospital-acquired pneumonia (HAP),hospitalization days in ICU,28-day mortality and adverse reactions of acupoint catgut embedding were observed in the two groups.Results After treatment,the levels of IAP,DAO,D-lactic acid were lower than those before treatment,GRV was less than that before treatment in the two groups,and after treatment for 7 days the changes were more significant in the observation group [IAP (cmH2O,1 cmH2O =0.098 kPa):12.42±3.11 vs.17.70 ± 2.98,GRV (mL):129.54 ± 57.54 vs.242.91 ± 99.29,DAO (U/L):9.06± 2.84 vs.13.60 ± 7.16,D-Lactate (mg/L):16.47 ± 5.39 vs.26.29 ± 9.62,all P ＜ 0.01].Days of mechanical ventilation (days:6.97 ± 4.13 vs.9.23 ± 4.61),time for reaching standard enteral nutrition (days:4.25 ± 3.15 vs.5.50 ± 3.12) and hospitalization days in ICU (days:9.50± 4.03 vs.12.20 ± 6.38) in observation group were significantly shorter than those in control group (all P ＜ 0.05).The incidence of HAP [15.6％ (5/32) vs.40.0％ (12/30)] and 28-day mortality [3.1％ (1/32) vs.6.7％ (2/30)] in observation group were obviously lower than those in control group.The patients in observation group had no significant adverse reactions of acupoint catgut embedding.Conclusions Acupoint catgut embedding therapy is not only easy to operate,but also can effectively improve gastrointestinal motility and intestinal barrier function in patients with gastrointestinal dysfunction after surgery of severe head injury,thus it is beneficial to reaching the enteral nutritional goal in less time,reducing the incidence of HAP,shortening the hospitalization time in ICU and promoting the recovery of the patients.

Objective To induce the experimental corneal neovascularization(CRNV) by alkali burn,and explore the methods for quantitative analysis of CRNV and observation of permeability of CRNV. Methods Thirty-six C57BL/6 mice were selected and divided into experiment group(n=30) and control group(n=6).For the experiment group,alkali burn was induced by application of filter paper with 1mol/L sodium hydroxide to the cornea for 5 s.For the control group,no intervention was conducted.Areas of CRNV were measured on day 4,7,10 and 14 after alkali burn.Histological examinations of cornea were performed with HE staining on day 3, 7,10,16 and 28 after alkali burn.On day 10,endothelial cell marker CD31 was used with immunohistochemical staining for CRNV counting,and fluorescence angiography(FA) was employed to reveal the permeability of CRNV.Corneal ulceration and hyphema were observed everyday.Results CRNV developed after alkali burn,and extincted afterwards.Axenic coneal ulceration and hyphema were frequently observed,with the incidences of 6.7% and 10.0%,respectively.Histologic changes of corneal tissues at different time points could be observed with HE staining.On day 10, CRNV could be labeled and counted with immunohistochemical staining for CD31 antibody,and the permeability of CRNV could be detected by FA. ConclusionCRNV counting with immunohistochemical staining for CD31 antibody and measurement of area of CRNV are appropriate methods for quantitative analysis of CRNV.FA is an effective method in the detection of permeability of CRNV.

AIM:To establish a liquid chromatography method for determining the monosaccharide composition of human lipoproteins, and to investigate the differences between diabetic patients and healthy participants.METHODS:Liquid chromatography with pre-column derivatization was used to determine the neutral and basic monosaccharides, and liquid chromatography tandem mass spectrometry was applied to quantify N-acetylneuraminic acid content.RESULTS:The contents of mannose, glucosamine, N-acetylglucosamine, glucose, galactose and N-acetylneuraminic acid in high-density lipoprotein from healthy participants and diabetic patients were (5.88 ±0.94),(16.49 ±4.11),(1.31 ±0.33), (0.87 ±0.16), (7.18 ±1.64), (2.14 ±0.12) mmol/(g protein) and (8.68 ±0.39), (24.73 ±5.50), (1.91 ±0.54), (1.23 ±0.35), (9.73 ±2.85), (3.53 ±0.27) mmol/(g protein), respectively.The contents of mannose, glu-cosamine,N-acetylglucosamine, glucose, galactose and N-acetylneuraminic acid in low-density lipoprotein from healthy par-ticipants and diabetic patients were ( 29.20 ±3.57 ) , ( 50.77 ±4.72 ) , ( 5.28 ±0.64 ) , ( 10.06 ±1.37 ) , ( 28.44 ± 3.96),(6.86 ±0.11) mmol/(g protein) and (30.08 ±3.78), (38.52 ±6.38), (3.79 ±0.78), (7.63 ±1.50), (20.05 ±2.63), (6.45 ±0.18) mmol/(g protein), respectively.The contents of mannose, glucosamine, glucose, ga-lactose and N-acetylneuraminic acid in very-low-density lipoprotein from healthy participants and diabetic patients were (91.21 ±4.12), (27.05 ±2.34), (4 230.95 ±15.83), (43.40 ±3.75), (2.95 ±0.24) mmol/(g protein) and (82.40 ±0.51), (30.16 ±0.32), (4 722.73 ±93.27), (34.05 ±2.81), (4.42 ±0.15) mmol/(g protein), respec-tively.CONCLUSION:Liquid chromatography with pre-column derivatization is suitable for the neutral and basic mono-saccharide analysis in human lipoproteins, and the glycosylation of lipoproteins in diabetic patients are significantly changed compared with the healthy controls.

Objective To investigate expression and significance of matrix metalloproteinase-2(MMP-2),MMP-9 and tissue inhibitor of metalloproteinase-1(TIMP-1) in rats with glomerular sclerosis made by doxorubicin.Methods Forty Wistar male rats(8-week-old) were randomly assigned into 2 groups:sham operated and model groups.Rats in model group were nephrectomized after anesthesia and injected with adriamycin(5 mg/kg) after 1 week.Rats in sham operated group was subjected to sham operation and injected with normal saline after 1 week through the tail vein.All rats were killed in the 12th week.Immuno-histochemistry was performed on renal tissue to detect Collagen Ⅳ(Col-Ⅳ),fibronectin(FN),MMP-2,-9 and TIMP-1.Results Immunohistochemistry staining indicated that expressions of MMP-2,-9 in model group decreased significantly compared to sham operated group(Pa

Objective To study the expression of ?-smooth muscle actin(?-SMA)in glomerulosclerosis rats and its relationship with renal function.Methods Forty healthy Wistar rats were equally divided into 2 groups including sham operated group and model control group.Rats in model groups were uninephrectomized and injected with daunorubicin(5 mg/kg)after 1 week through the tail vein.Twenty-four hours of urinary protein excretion,serum creatinine(Scr),blood urea nitrogen(BUN)were measured at the 12th week.Renal pathology was evaluated.Immunohistochemistry(SupervisionTM)was performed on renal glomeruli tissue to detect the expression of ?-SMA.Reverse transcription polymerase chain reaction(RT-PCR)was used to examine the expression levels of ?-SMA mRNA in glomeruli.SPSS 13.0 software was used to analyze the two variables.Results In model control group,the urinary protein,Scr,BUN significantly increased(Pa