Background Carbachol solution (0.01％) is an agonist of M cholinoceptor and skeletal muscle N cholinoceptor,and it is used to play miotic effect and open peripheral iridectomic hole during the surgery of implantable collamer lens (ICL) implantation in order to lower intraocular pressure (IOP).However,the anterior chamber injection of 0.01％ carbachol solution often causes relevant complications,while whether lower dose of carbachol solution is effective and safe is unclear.Objective This study was to compare the effectiveness and safety between 0.01％ carbachol solution and 0.005％ carbachol solution after anterior chamber injection during the ICL implantation.Methods One hundred and fifty-two eyes of 76 cataract patients were included in Daping Hospital of Third Military Medical University from September 2014 to September 2015.ICL implantation and periphery iridectomy were carried out on both eyes of the patients and the 0.01％ carbachol solution was injected into the anterior chamber during the surgery of the right eyes and 0.01％ carbachol solution was used in the left eyes.The operation duration and IOP at postoperative 2 hours and systemic choline-like reaction were compared between 0.01％ carbachol solution group and 0.005％ carbachol solution group.Results The mean operation duration was (11.86± 2.39) minutes and (11.22 ± 1.85) minutes in the 0.01％ carbachol group and 0.005％ carbachol group,without significant difference between two groups (t =1.851,P =0.066).IOP was (15.76 ± 2.18) mmHg and (13.58 ±2.24)mmHg in the 0.01％ carbachol group before and after surgery,and those in the 0.005％ carbachol group was (15.70±2.35)mmHg and (13.12±2.17)mmHg,there was no significant difference in the IOP between the two concentrations of carbachol (Fsroup =0.986,P=O.322).The IOP at postoperative 2 hours was lower than that before operation,the difference was statistically significant(Ftime =97.339,P =0.000).There was no interaction between drug concentration and time (Fcorrelation =0.772,P =0.381).The incidences of complications,such as dizziness,nausea and vomiting,were lower in the 0.005％ carbachol group than those in the 0.01％ carbachol group (x2 =13.01,5.16,4.03,all at P＜0.05).Conclusions Carbachol solution (0.005％) can play intraoperative miosis effect and maintain effective operation duration in ICL implantation.In addition,the application of 0.005％ carbachol solution is quite safe in both intraoperation and postoperation.

Objective To describe the phenotype of NK cells in Bama miniature pigs, and establish an efficient activation and culture method for porcine cytokine-induced killer ( CIK) cells in vitro. Methods The porcine peripheral blood mononuclear cells ( PBMCs) were isolated by Percoll gradient centrifugation, and the phenotype of NK cells was test-ed by detecting the CD2 + /CD8 + /CD3 - cell compartment. To establish an efficient activation and culture method for por-cine CIK cells, we optimized the culture conditions to improve the CIK activation efficiency. Results Using the optimized induction culture conditions, the ratio of CIK ( CD2 + /CD8 + /CD3 -) cells was up to 43. 63% at the fifth day, approxi-mately 5. 59 times increased compared with the initially separated PBMCs. Cell proliferation experiments showed that three obvious fluorescence peaks were observed on the fifth day. The results indicated that the induced CIK cells underwent three times cell division, in theory, about increased 8-fold compared with the initial separation of PBMCs. Furthermore, the qRT-PCR result of the surface markers of porcine NK cells also showed a similar variation tendency as the flow cytometry results. Conclusions Our findings demonstrate the successful establishment of an efficient activation and culture method for porcine CIK cells in vitro.

Objective To investigate the clinicopathological characteristics of HBV recurrence after liver transplantation. Methods The retrospective analysis of the clinicopathological changes was performed on 17 patients who had HBV recurrence after liver transplantation in our medical department. Results HBV recurrence happened from 4 to 48 months. Twelve of them which were identified to be YMDD mutation switched to entecavir or added adefovir. Three of them receiving chemotherapy when liver cancer recurred switched to entecavir. Two of them with withdrawal of lamivudine were given lamivudine continuously. Liver function returned to the normal level and HBVDNA was ＜ 102 U/ml after anti-hepatitis B virus. The histological changes in the transplanted livers included hepatocellular degeneration, necrosis and apoptosis, portal infiltrations and fibrosis.With time after recurrence, it was easier to see hepatitis B virus replication in liver cells, incidence of acute rejection, increases of liver fibrosis and the formation of fibrous septa, even pseudolobule.Conclusion In native HBV infection livers, fibrosis occurs more early and develops rapidly. The number of virus is closely related to liver necrosis and inflammation. Early discovery and change to quick and effective treatment of anti-hepatitis B virus in time can improve greatly the prognosis of the patients.

Objective To investigate the effect of microcireulation blood flow and altering immunity by Chuanxiongqin,ulinastain and thymosin α1 on sepsis patients.Methods 90 patients were randomly divided into 3 groups(n=30),namely ICU group,Chuanxiongqin group,ulinastain and thymosin α1 group.HLA-DR/CD14+and IL-6,TNF-α,Lac,DD were measured.Results (1)DD showed no significant difference at every time point between ICU group and ulinastain+thymosin α1 group(P＞0.05).DD decreased in Chuanxiongqin group,and was significantly different from the others on the third day.(2)Lac unchanged significantly at every time point in ICU group(P＞0.05).Lac in Chuanxiongqin group and ulinastain+thymeain α1 group tended to decrease,and was statistically different from ICU group on the second day.(3)IL-6 and TNF-α tended to increase at every time point in ICU group(P＜0.05).In ulinastain+thymosin α1 group,IL-6 and TNF-α returned to the level before treatment,HLA-DR/(D14+increased significantly,and was higher than Chuanxiongqin group and ICU group statistically.Conclusion Chuanxiongqin could ameliorate circulation;ulinastsin and thymosin α1 could depress IL-6,TNF-α.So ulinastain and thymosin α1 might protect the immunity of sepsis patients.

Objective To develop a real-time RT-PCR assay ( rRT-PCR) for efficient detection of duck hepatitis virus type 1 ( DHV-I) .Method According to the different gene sequences of DHV-I from different provinces download from NCBI and to find the conserved sequences.One pair of the specific primers and one TaqMan probe were designed. Then reaction parameters were optimized to develop a real-time RT-PCR assay ( rRT-PCR) .Results This developed rRT-PCR assay could detect 20 template copies of RNA, and its sensitivity was higher than that of the conventional RT-PCR. This rRT-PCR assay was found to be specific and able to detect DHV-I, and no positive results were observed when nucleic acid from Muscovy duck parvovirus, goose parvovims, Newcastle disease and avian influenza virus, egg drop syndrome virus, reticuloendotheliosis virus, duck Tembusu virus, poultry intestinal arc virus were used as rRT-PCR templates.The results of this developed rRT-PCR assay used for 100 duck clinical samples showed a positive rate of 92%, indicating that DHV exists in duck group of Jiangsu province in China.Conclusion This rRT-PCR assay can be used as a rapid tool for detection of DHV-I.

OBJECTIVETo establish a SPE-HPLC method for analyzing astilbin in rats serum and explore the effects of Yinxieling (YXL) prescription and Smilacis Glabrae Rhizoma on the pharmacokinetic characteristics of effective components.

METHODMale Sprague-dawley rats were administrated YXL and Smilacis Glabrae Rhizoma respectively. At different time points, serum concentration of astilbin was extracted by Solid Phase Extraction (SPE) and determined using HPLC method. Chromatographic separation was achieved on a reversed-phase Phenomenex C18 column with the mobile phase of methanol-acetonitrile-formic acid water solution (0.05% formic acid) and gradient elution, temperature of bar was 24 degrees C, flow rate was 0.8 mL x min(-1).

RESULTThe method showed excellent linearity over the concentration range 0.266-53.1 mg x L(-1) of astilbin (r = 0.996). The extract recoveries were from 79.0% to 89.1%. Significant diffenerce in pharmacokinetic parameter of astilbin including t1/2, Cmax, AUC(0-t), AUC(0-infinity) and MRT were obtained through non-compartment model after oral administration of YXL prescription comparing with Smilacis Glabrae Rhizoma.

CONCLUSIONThe method was applied to a pharmacokinetic study of astilbin. It indicated that the bioavailability of astilbin in YXL enhanced in rats and one of the reasons may be that components of prescription affect the pharmacokinetics of astilbin in vivo.

Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Flavonols ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results

During severe acute respiratory syndrome coronavirus (SARS-CoV) infection, the activity of the replication/transcription complexes (RTC) quickly peaks at 6 hours post infection (h.p.i) and then diminishes significantly in the late post-infection stages. This "down-up-down" regulation of RNA synthesis distinguishes different viral stages: primary translation, genome replication, and finally viron assembly. Regarding the nsp8 as the primase in RNA synthesis, we confirmed that the proteolysis product of the primase (nsp8) contains the globular domain (nsp8C), and indentified the resectioning site that is notably conserved in all the three groups of coronavirus. We subsequently crystallized the complex of SARS-CoV nsp8C and nsp7, and the 3-D structure of this domain revealed its capability to interfuse into the hexadecamer super-complex. This specific proteolysis may indicate one possible mechanism by which coronaviruses to switch from viral infection to genome replication and viral assembly stages.
Amino Acid Sequence ; Crystallography, X-Ray ; DNA Primase ; chemistry ; genetics ; physiology ; Humans ; Isoenzymes ; chemistry ; genetics ; physiology ; Molecular Sequence Data ; Protein Structure, Secondary ; RNA, Viral ; biosynthesis ; SARS Virus ; chemistry ; genetics ; physiology ; Sequence Alignment ; Severe Acute Respiratory Syndrome ; virology ; Virus Replication

Acupuncture can improve the motor and non-motor symptoms of Parkinson's disease, and the effect of acupuncture combined with drug therapy is better than that of drug therapy alone. The possible mechanism includes inhibiting α-synuclein aggregation, oxidative stress, and neuroinflammation, inhibiting the apoptosis of dopaminergic neurons, and achieving a neuroprotective effect. The points mainly selected for Acupuncture treatment for this disease are Zusanli (ST 36), Yanglingquan (GB 34), Taichong (LR 3), Xuehai (SP 10), and other points. Early use of acupuncture and acupuncture combined with medical treatment strategy is worthy of clinical application.