A strain of Mucor named M2,which could produce protease,was isolated from a traditional fermented soybean product.Culture conditions of proteases produced by M2 were studied therefore.The results showed that nitrogen source and carbon source preferred for protease production were soybean protein isolate and glucose,while inorganic salts preferred were KH2PO4,CaCl2 and MgCl2.The suitable culture conditions for protease production were as follows:culture temperature was 28℃,inoculation volume was 2%,liquid level was 100 mL in 300 mL triangle bottle at pH 5,rotating speed was 150 r/min and culture time was 48 h.The obtained protease activity in culture was about 4.35 U/mL.The protease produced by Mucor was analyzed with SDS-PAGE.The protease had a molecular weight of 36.4 kD.

OBJECTIVETo compare the difference in the clinical efficacy on posterior circulation ischemia between acupuncture at stellate ganglion and conventional acupuncture as well as the impacts on blood pressure.

METHODSEighty cases of posterior circulation ischemia were randomized into an observation group (40 cases) and a control group (40 cases). In the observation group, acupuncture was applied to the bilateral stellate ganglions on the neck, stimulated with reinforcing technique by rotating the needles. In the control group, the acupuncture of reducing technique was applied to Fengchi (GB 20), Baihui (GV 20), Neiguan (PC 6) and Taichong (LR 3) in the excess syndrome. The even needling or reinforcing technique was applied to Fengchi (GB 20), Baihui (GV 20), Ganshu (BL 18), Shenshu (BL 23) and Zusanli (ST 36) for the deficiency syndrome. The treatment was given once every 3 days and 4 treatments were required totally in the two groups. The changes in total syndrome score, peak Systolic blood flow velocity (Vp) of vertebral artery and basilar artery, systolic and diastolic blood pressures were compared before and after treatment in the two groups. The clinical efficacy was compared between the two groups.

RESULTSThe total syndrome score was reduced apparently after treatment compared with that before treatment in the two groups (P < 0.01), and the reducing was more obvious in the observation group as compared with that in the control group (P < 0.01). The total effective rate was 87.5% (35/40) in the observation group, higher than 67.5% (27/40, P < 0.05) in the control group. After treatment, the reduced Vp of vertebral artery was not improved apparently as compared with that before treatment in the control group, Vp in blood velocity abnormality (including vascular spasm, stenosis or reduced velocity) of vertebral artery and basilar artery was all improved as compared with that before treatment in the two groups (P < 0.01), and the improvements in the observation group were more obvious than those in the control group (P < 0.01). After treatment, the systolic and diastolic pressures were reduced as compared with those before treatment in the two groups, and the reduced systolic and diastolic pressures in the observation group were more apparent than those in the control group (P < 0.01).

CONCLUSIONAcupuncture at stellate ganglion achieves the satisfactory efficacy in the treatment of posterior circulation ischemia and the significant efficacy of reducing blood pressure, more advanced than the conventional acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Blood Pressure ; Brain Infarction ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stellate Ganglion ; physiopathology ; Treatment Outcome

A novel hydrophobic ferrocene polymer ( P-Fc) was synthesized successfully. Taken the advantage of 2-hydroxypropyl-β-cyclodextrin ( HP-β-CD ) with hydrophilic outside and hydrophobic inside, P-Fc was successfully enveloped in the cavity of HP-β-CD to form macromolecular vesicle by a self-assembly process. The structure and morphology of the macromolecular vesicle was characterized by FTIR, 1HNMR, SEM and CV. Rhodamine 6G (R6G) and doxorubicin hydrochloride (DOX) was used as target drugs and loaded in the vesicles successfully. After added the oxidant, ferrocene was oxidized to dicyclopentadienyl iron, which destroyed the vesicles successfully. The rapid directional release of the drugs was realized. The maximum loading values of R6G and DOX were 6. 89 and 39. 06 μg/mg, respectively. The release rates were 73. 7%and 88. 2%, respectively. It would increase the utilization level of the drugs and enhance their efficiency.

[Ab]stract] Objective To investigate, according to core myodynamic theory, the clinical efficacy of electroacupuncture at Huatuo jiaji points in treating motor dysfunction in stroke patients during the flaccid paralysis stage.Methods Sixty stroke patients in the stage of flaccid paralysis were randomly allocated to treatment and control groups, 30 cases each. The control group received basic treatment and conventional acupuncture and the treatment group, electroacupuncture at Huatuo jiaji points in addition. The simplified Fugl-Meyer assessment (FMA) score and the Modified Barthel Index (MBI) score were counted before and after treatment and the clinical therapeutic effects were compared between the two groups.Results There were statistically significant pre-/post-treatment differences in the FMA score and the MBI score in the two groups (P<0.05). There were statistically significant post-treatment differences in the FMA score and the MBI score between the treatment and control groups (P<0.05). The total efficacy rate was 96.7% in the treatment group and 86.7% in the control group; there was a statistically significant difference between the two groups (P<0.05).Conclusion Electroacupuncture at Huatuo jiaji points has a promoting effect on the rehabilitation of limb function in the flaccid paralysis stage of stroke.

Objective To observe the clinical efficacy of thread embedding at Back-Shu points in treating stroke-associated pneumonia.Method Seventy patients with stroke-associated pneumonia were randomized into a treatment group and a control group, 35 cases in each group. The control group was intervened by regular medications, while the treatment group was additionally intervened by thread embedding at the Back-Shu points. Before and after the intervention, C-reactive protein (CRP), white blood cell (WBC), neutrophil ratio (NE%), and symptoms and signs scores were observed, and the clinical efficacies were compared between the two groups.Result The CRP, WBC, NE%, and symptoms and signs scores were significantly changed after intervention in both groups (P<0.05). After intervention, the CRP, WBC, NE%, and symptoms and signs scores in the treatment group were significantly different from that in the control group (P<0.05). The total effective rate was 91.4% in the treatment group, versus 82.9% in the control group, and the difference was statistically significant (P<0.05).Conclusion Thread embedding at the Back-Shu points can promote the treatment of stroke-associated pneumonia.

Objective To investigate the therapeutic effect of high intensity focused ultrasound (HIFU) ablation combined with homemade liquid fluorocarbon nanoparticles on cervical cancer in nude mice.Methods The cell experiment was divided into three groups:a,control group;b,HIFU group;c, HIFU + PFB nanoparticle group,and the viability of cells was detected using CCK-8 reagent.The mice were also divided into three groups:A,0.9% NaCI group;B,HIFU + 0.9% NaCI group;C,HIFU + PFB nanoparticle group. The tumors were removed and underwent triphenyl tetrazolium chloride(TTC) staining,and the necrosis area was measured.Histopathological changes of the tumors were examined by light microscopy.Results After HIFU irradiation,the viability rate of group c was (40.5 ±9.7)%,it was lower than that of group b (77.7 ±8.5)% (P <0.05) and that of group a(100 ±4.8)% (P <0.05). TTC staining of tumor showed a large scale of necrotic tissue in group C.The necrosis ratio of the three groups was 0%,(34.14±12.2)% and (65.97 ±25.1)%,respectively (P <0.05).HE staining showed karyorrhexis or an absence of nuclei in group B and group C,which demonstrated the coagulation necrosis. Conclusions HIFU ablation combined with liquid fluorocarbon nanoparticles can effectively treat the xenograft model of the human cervical carcinoma in nude mice.

Objective To compare the difference of the clinical efficacy on post-stroke shoulder-hand syndrome treated by acupuncture at stellate ganglion and by conventional acupuncture. Methods Ninety cases of post-stroke shoulder-hand syndrome ( at stageⅠ or stageⅡ) were randomized into treatment group and control group, 45 cases in each group. Both groups received basic rehabilitation training. Additionally, the treatment group received acupuncture at the bilateral stellate ganglions of the neck, with reducing acupuncture by rotating the needles. The control group received the conventional acupuncture on Jiquan ( HT 1) , Chize ( LU 5) , Jianyu (LI 15), Quchi (LI 11), Shousanli (LI 10), Hegu (LI 4), with reducing acupuncture by lifting and thrusting the needle on Jiquan ( HT 1) and Chize ( LU 5) , and with mild reinforcing and reducong acupuncture on the other acupoints. The treatment for both groups was given once every day, and 14 times in total. The total syndrome scores, visual analogue scale (VAS) scores, modified Fugl-Meyer Assessment of motor function scale (FMA) scores, and modified Barthel index (MBI) were observed to evaluate the changes of symptoms, pain and motor function of the upper limbs, and activity of daily life before and after treatment in the two groups. The clinical efficacy was also compared between the two groups. Results The total syndrome scores, VAS scores, FMA scores and the modified Barthel index were improved apparently after treatment in the two groups (all P<0.01), and the improvement in the treatment group as more obvious (all P<0.01) . The total effective rate was 93.33% (42/45) in the treatment group, which was better than 73.33% (33/45) in the control group ( P<0.05) . During the treatment, patients of both groups had good compliance, and all completed the trial. No obvious adverse reaction was found in the two groups. Conclusion Acupuncture at stellate ganglion is effective and safe for the treatment of post-stroke shoulder-hand syndrome, and it has higher efficiency than the conventional acupuncture.

AIM: To evaluate the efficacy of combined therapy for amblyopia in children by making use of pattern visual evoked potential ( P-VEP) game.
METHODS: This was a prospective case control study. These asthenopic children were divided into two groups. The control group ( 66 eyes of 49 patients ): occlusive therapy with glasses, cover, precision work, red light treatment and so on, later the stereo vision training was added. The experimental group (72 eyes of 52 patients):conventional methods mentioned above with P - VEP games.
RESULTS: The total effective rate and cure rate of experimental group in 6mo were higher than those of control group. The overall effective rate was 94. 4% in the experimental group and 83. 3% in the control group. There was a statistically significant difference between them (P<0. 05).
CONCLUSION: The comprehensive therapy by making use of P-VEP game is an individualized effective new way in treating amblyopia.

Objective To investigate the clinical features and LRRK2 gene mutation in patients with autosomal dominant familial Parkinson's disease (PD). Methods The clinical features of 16 autosomal dominant familial PD probands were analyzed in terms of age at onset, onset symptoms, UPDRS scores, response to the levodopa treatment and drug-induced dyskinesia. The LRRK2 gene exons 5,13,31,32,35,37,41 and 48 of 16 probands were sequenced after polymerase chain reaction. The novel mutation was further screened in 24D sporadic PD patients and 214 controls using PCR-RFLP for the genotypo frequency analysis. Results Clinically, most of 16 probands had late-onset age. Resting tremor (9patients, 56. 25%,t=0.558,P=0.679)and bradykinesia (9 patients,56.25%,t=0.369,P=0.454)were common onset symptoms followed by rigidity(6 patients,37.50%,t=1.324,P=0.735)and postural instability(5 patients,31.25%,t=2.369,P=0.956).Majority of them had good response to levedopa treatment and rare occurrence of drug-induced dyskinesia. Among the 16 autosomal dominant familial PD probands,6 variants were identified:c.457 T＞C(Leu153Leu),c.1432 G＞T(Asp478Tyr),c.5457 T＞C(Gly1819Gly),c.7153 G＞A(Gly2385Arg),IVS31+28 T＞G and IVS37+162 T＞C. The c.1432G＞T(Asp478Tyr)variant was a novel mutation and it was not detected in 240 sporadic PD patients and 214 controls. The reported mutations associated with the PD, such as Arg1441 Cys/Gly/His, Arg1514Gln, Tyr1699Cys, Ile2012Thr, Gly2019Ser and Ile2020Thr,were not found in our study. Conclusions The autosomal dominant familial PD patients present with classical symptoms of PD and bear the LRRK2 variantsAsp478Tyr and Gly2385Arg.

OBJECTIVETo compare the effect of reinforcing Shen method (RSM) and activating blood method (ABM) in treating osteoarthritis (OA) at the molecular level.

METHODSThe physical and chemical characteristics of components from respective recipes of RSM and ABM, and network features of component-target interaction network were analyzed by computer simulation methods including chemical space, molecular docking, and biological network, etc.

RESULTSThe chemical components of RSM and ABM were scarcely scattered with larger overlapping. Among established networks, the distribution of network features was partially similar in RSM and ABM. The average target number correlated with each component was 1.86 in RSM and 2.11 in ABM respectively. Each average target number was respectively correlated with 4.46 compounds and 3.93 compounds, reflecting multi-component and multi-target actions.

CONCLUSIONComputer simulation could intuitively trace out similarities and differences of two different methods and their interaction with targets, which revealed that the compatibility of RSM and ABM could have broader protein targets and potential synergism at the molecular level.

Computer Simulation ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Osteoarthritis ; drug therapy ; Phytotherapy ; methods