OBJECTIVETo study organic anion transporting polypeptide (OATP) superfamily member 4a1 (oatp4a1) mRNA expression in the Pi deficiency model rats, thus exploring its mechanism for transporting and transforming the dampness.

METHODSSix SD rats of SPF grade were used to prepare over-fatigue impairing Pi model. Another 12 SD rats were randomly divided into the blank control group and the high fat diets group, 6 in each. The special binding tube was used for the over-fatigue impairing Pi model group on the odd day, 3 h each time. Then the rats were forced to swim in the cold water (10 degrees C +/- 1 degrees C) for 7 min on the even day, for 2 successive weeks. Rats in the model group and the blank control group were granulated feed for 12 weeks, while rats in the high fat group were fed with high fat diet for 12 weeks. All rats were free to take food and drink water. The mRNA and protein expressions of oatp4al were detected in the Fei, Pi, Gan, Shen, Wei, Xiaochang, and Dachang using Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blot.

RESULTSRats in the model group were idled together with lousy defecate and coarse skin. They ate and drank less, and lost body weight (P<0.05). They were consistent with clinical manifestations of Pi deficiency syndrome, indicating that the over-fatigue impairing Pi animal model was successfully established. Rats in the high fat group started to have poor appetite and languish spirit, move lazily and addict to sleep, have coarse, dark, and colorless hair 9 weeks later, indicating phlegm dampness syndrome. Compared with the blank control group, the average body weight increased in the high fat group at the 9th week (P<0.05). The oatp4a1 mRNA expressed in the Fei, Pi, Gan, Shen, Wei, Xiaochang, and Dachang. There was no statistical difference in the oatp4al mRNA expression among all tissues (P>0.05). The oatp4al mRNA expressions were higher in the Fei and Shen of the high fat group than in the Gan (P<0.05).

CONCLUSIONSoatp4al might be one of the basic substances in the transportation and transformation of phlegm dampness. Of them, Fei, Shen, and Dachang might play important roles in the transportation and transformation of phlegm dampness.

Animals ; Antiporters ; metabolism ; Diet, High-Fat ; Eye Proteins ; metabolism ; Fatigue ; metabolism ; Male ; Organic Anion Transporters ; metabolism ; RNA, Messenger ; genetics ; Rats ; Yin Deficiency ; metabolism

Objective To investigate the efficacy of Ilizarov technique in treatment of Gustilo type Ⅲ B and C of large tibia bone defect combined with soft tissue defect. Methods Thirty cases with large tibia bone defect combined with soft tissue defect in our hospital from January 2016 to December 2018 were selected including 25 cases with type Ⅲ B and 5 cases with type Ⅲ C,who were all open tibia fractures and treated with Ilizarov technique. Results All patients were followed up for 10 to 18 months with an average of 15 months. The bone defects were reconstructed, the difference between the length of the affected limb and the healthy side was less than 2 cm. The fracture was healed and the wound was closed. One case was with paralysis of common peroneal nerve and recoveried after 3 months. Twenty-two cases recovered without extra surgery, five cases restored with skin graft and three cases covered the wound by rotating skin flap. Conclusion Ilizarov technique is one of effective ways to treat Gustilo Ⅲ type B and type C of large defect combined with soft tissue defect.

Objective: To learn the prevalence and drug resistance of nontuberculous mycobacteria ( NTM ) in Zhejiang Province, so as to provide evidence for NTM prevention and control. Methods: A total of 2 878 clinical mycobacterium isolates in Zhejiang Province were collected from the drug resistance surveillance in 2008-2009, 2013-2014 and 2018-2019, PNB/TCH growth tests were used to preliminarily identify the NTM in these mycobacterium-positive isolates. 16SrRNA, rpoB, ITS and hsp65 gene sequencing analysis were used to confirm strains initially identified as NTM. Proportional method was applied to detect drug susceptibility of NTM isolates. Results : Finally, 135 strains were confirmed as NTM and the isolation rate was 4.69%. The isolation rates of NTM in 2008-2009, 2013-2014 and 2018-2019 were 1.85%, 4.56% and 7.84%, respectively, with an increasing trend ( P<0.05 ). Thirteen species were identified and the top two species were M. intracellulare ( 82, 60.74% ) and M. kansassi ( 18, 13.33% ). The NTM isolates showed the highest drug resistance rate to isoniazid ( 97.78% ), followed by p-aminosalicylic acid ( 94.87% ) and streptomycin ( 94.81% ). Conclusions The isolation rates of NTM showed an upward trend in the drug resistance surveillance in 2008-2019,2013-2014 and 2018-2019 in Zhejiang Province. M. intracellulare and M. kansassi were the main strains isolated. The NTM isolates showed high resistance against both first and second-line antituberculosis drugs.

Objective To know the application of MPB64 antigen detection of immune colloidal gold method for identification of Mycobacterium.Methods Taking the PCR amplified sequencing method as the 'gold standard',a total of 418 clinical isolated mycobacterium tuberculosis and 2 standard strains were tested by immune colloidal gold method,and was compared with the traditional method of PNB/TCH.Results Compared with sequencing method,the coincidence rate of immune colloidal gold method and PNB/TCH identification test was 95. 93%and 87. 08%respectively.The sensitivity, specificity,positive predictive value (PPV ) and negative predictive value (NPV ) of immune colloidal gold method identification were 88. 41%,99. 64%,99. 19% and 94. 58%respectively.It costs MPB64 immune colloidal gold method 15 minutes to carry out the results,less than 28 days of the traditional method,as well as 3 days of sequencing method. Conclusion Compared with PNB/TCH method,MPB64 immune antigen colloidal gold method has higher specificity and sensitivity,and could be applied in identification of Mycobacterium.

OBJECTIVETo construct a gene knock-out mutant of response regulator named RevS in Streptococcus suis serotype 2 virulent strain 05ZYH33, and to investigate the effects of its deletion on the biological characters of this pathogen and the pathogenesis to mice and piglets.

METHODSRecombinant gene knock-out vector consisting of Spc(r) cassette was constructed and flanking was constructed consisting of Spc(r) cassette with flanking homology regions to the RevS genes while the isogenic RevS-deficient mutant was screened by allelic replacement. The effects of RevS deletion on the basic biological characters of 05ZYH33 including growth stability, colonial morphology, haemolysis, Gram staining, growth curve and protein expression were examined in vitro. The mice and piglets were infected with 10(8) CFU wild virulent and mutant isolates.

RESULTSPCR analysis confirmed that the coding genes of RevS were replaced completely by Spc(r) cassette and the basic biological characters of 05ZYH33 did not undergo any apparent change. Balb/c mice infection assay indicated that RevS play a role in the pathogenesis of Streptococcus suis infections, while no remarkable difference was observed in the piglets' pathogenesis infection rates between mutant isolates deltaA05ZYH33 and wild-type isolates 05ZYH33.

CONCLUSIONThe mutant of Streptococcus suis 05ZYH33 response regulator was successfully constructed, while the mutation did not obviously affect the bacterial biological characters, while the knock-out mutant of RevS was shown to be attenuated in pathogenesis to mice and piglets.

Animals ; Bacterial Proteins ; genetics ; Gene Knockout Techniques ; methods ; Mice ; Mice, Inbred BALB C ; Models, Genetic ; Polymerase Chain Reaction ; Streptococcal Infections ; microbiology ; Streptococcus suis ; genetics ; pathogenicity

OBJECTIVETo determine the prevalence of Streptococcus suis and major pathogenic serotypes in middle part of Jiangsu province.

METHODSTonsillar specimens from 303 slaughtered pigs aged 6 to 8 months were investigated for the presence of Streptococcus suis and major pathogenic serotypes by polymerase chain reaction (PCR) method. Bacteriological examination compared with molecular genetics identification for three Streptococcus suis isolates were also done.

RESULTSThe overall carrier rate of Streptococcus suis was up to 88.0%, with the percentages of serotype 1(14), 2(1/2), 7 and 9 were 9.6%, 8.5%, 11.3% and 29.5% respectively in 2005. While in 2006, the prevalence of Streptococcus suis was 82.5%, with capsular types 1 (14), 2 (1/2), 7 and 9 were accounted for 17.6%, 2.4%, 25.8% and 20.0% of all the specimens. All the three isolates belonged to Streptococcus suis serotype 2,named 2a, 2f and 14e, which exhibiting the virulent phenotype cps2+/gdh+/mrp-/lepf-/sly-/fbps+/orf2+/89k-, cps2+/lgdh+/mrp-/epf-/sly-/fbps-/orf2-/89k- and cps2+/gdh+/mrp-/epf-/sly-/fbps/orf2-/ respectively. These isolates were all susceptible to amoxicillin, ampicillin, penicillin and resistant to amikacin and tetraycline. Clinical signs were not noted in BALB/c mice and rabbit.

CONCLUSIONPrevalence of the Streptococcus suis among the healthy herds in the areas was very high, with various capsule types of Streptococcus suis involved in the same herds, and the virulent phenotype of these 3 isolates were very different from those prevalent Streptococcus suis serotype 2 virulent isolates frequently discovered from the epidemic areas.

Amikacin ; therapeutic use ; Amoxicillin ; therapeutic use ; Ampicillin ; therapeutic use ; Animals ; China ; epidemiology ; Mice ; Mice, Inbred BALB C ; Molecular Epidemiology ; methods ; Penicillins ; therapeutic use ; Polymerase Chain Reaction ; Streptococcal Infections ; drug therapy ; epidemiology ; microbiology ; Streptococcus suis ; classification ; drug effects ; genetics ; pathogenicity ; Tetracycline ; therapeutic use ; Virulence

OBJECTIVETo explore roles of mRNA and protein expressions of organic anion transporting polypeptide (oatp2b1) of rats with high fat diet and overstrain induced Pi deficiency syndrome in the transporting of damp turbidity.

METHODSTotally 24 SD rats were randomly divided into three groups, i.e., the normal group, the overstrain group, and the high fat diet group, 8 in each group. After successful modeling, one piece of tissues such as spleen, kidney, liver, lung, stomach, small intestine, and large intestine was taken from each rat. Rats of the overstrain group were bonded by specially made bondage cylinder, 3 h each time on odd days, and forced to swim in cold water (10 +/- 1) degrees C for 7 min on even days alternatively for twelve weeks. Rats in the model group and the normal group were fed with standard routine granular forage for 12 weeks. Rats in the high fat diet group were fed with high fat forage for twelve weeks. All rats drank and ate freely. The mRNA and protein expressions of oatp2b1 were detected in the seven tissues using RT-PCR and Western blot.

RESULTSThe mRNA expression of oatp2b1 in liver and kidney tissues of rats in the high fat diet group was higher when compared with that of the normal group and the overstrain group (P < 0.01, P < 0.05). The oatp2b1 mRNA expression in the normal group was sequenced from high to low as liver > lung > spleen > larger intestine > small intestine > kidney > stomach. The oatp2b1 mRNA expression in the overstrain group was sequenced from high to low as liver > lung > larger intestine > spleen > kidney > stomach > small intestine. The oatp2b1 mRNA expression in the high fat diet group was sequenced from high to low as liver > lung > spleen > small intestine > kidney > larger intestine > stomach. The oatp2b1 protein expression in the lung tissue was sequenced from high to low as the overstrain group > the normal group > the high fat diet group (P > 0.05). The oatp2b1 protein expression in the spleen tissue was sequenced from high to low as the high fat diet group > the normal group > the overstrain group (P > 0.05). The oatp2b1 protein expression in the kidney tissue was sequenced from high to low as the normal group > the overstrain group > the high fat diet group (P > 0.05). The oatp2b1 protein expression in the liver tissue was sequenced from high to low as the normal group > the high fat diet group > the overstrain group (P > 0.05). Of them, the oatp2b1 protein expressed extremely less in the stomach, large intestine, and small intestine. The oatp2b1 protein expression in the normal group was sequenced from high to low as lung >spleen > liver, kidney > stomach, larger intestine, and small intestine. The oatp2b1 protein expression in the overstrain group was sequenced from high to low as lung > spleen > kidney > liver > stomach, larger intestine, and small intestine. The oatp2b1 protein expression in the high fat diet group was sequenced from high to low as spleen > lung > kidney > liver > stomach, larger intestine, and small intestine. However, there was no statistical significance among the three groups by pair-wise comparison (P > 0.05).

CONCLUSIONSKidney and liver might play important roles in the transportation and transformation of damp under the state of Pi deficiency syndrome. Oatp2b1 may be one of the material bases involved in the transportation and transformation of damp turbidity. Pi's function of governing transportation and transformation of damp might not only include the functions of the gastrointestinal tract, but also include partial liver and kidney functions.

Animals ; Diet, High-Fat ; Disease Models, Animal ; Fatigue ; diagnosis ; metabolism ; Kidney ; metabolism ; Liver ; metabolism ; Male ; Medicine, Chinese Traditional ; Organic Anion Transporters ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

BACKGROUNDTraumatic brain injury (TBI) is a major cause of death and disability in children and young adults worldwide. Therefore, we investigated the role of AG490 in regulating brain oedema, expression of CD40 and neurological function after TBI.

METHODSSprague Dawley rats (n = 240) were randomly divided into a sham operation group, TBI+saline group and TBI+AG490 (JAK/STAT inhibitor) group. Members of each group were euthanized at 6, 12, 24 or 72 hours after injury. Neurological severity score (NSS) was used to evaluate the severity of neurological damage. Brain water was quantitated by wet/dry weight method. The expression of CD40 was assessed by flow cytometry.

RESULTSIn both the TBI+saline group and the TBI+AG490 group, the brain water content was elevated after TBI, reached a peak at 24-hour and remained high for the rest of the period investigated; the expression of CD40 reached a peak 24 hours after TBI; the NSS was elevated after TBI and then decreased after 6 hours. Elevations in the level of CD40, degree of brain edema and NSS after TBI were significantly reduced in TBI+AG490 group.

CONCLUSIONInhibition of the JAK/STAT signalling pathway reduces brain oedema, decreases the expression of CD40 and exerts neuroprotective effects after TBI.

Animals ; Brain Edema ; metabolism ; Brain Injuries ; drug therapy ; CD40 Antigens ; analysis ; Flow Cytometry ; Janus Kinases ; metabolism ; Male ; Neuroprotective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; STAT Transcription Factors ; metabolism ; Tyrphostins ; therapeutic use

OBJECTIVETo observe the effect of electroacupuncture on recovery of urinary bladder function after radical hysterectomy.

METHODSOne hundred and ten cases were randomly divided into an electroacupuncture (EA) group and a control group, 55 cases in each group. In the control group, the urinary tube was placed and kept with routine method and the urinary bladder was rinsed, and from the eighth day the abdomen was radiated with TDP, 30 min each day, for 5 days. In the EA group, on the basis of treatment in the control group EA was given at Sanyinjiao (SP 6), Zusanli (ST 36), Waiguan (TE 5), Shuidao (ST 28), Guilai (ST 29), etc. from the eighth day to twelfth day after operation. The recovery time of urinary bladder function after radical hysterectomy, urine dynamic indexes and hospitalization days were compared between the two groups.

RESULTSThe cases of the bladder function recovery, retention of urine, urinary incontinence were 51(51/55), 4(4/55), 0 on the 14 th day after operation and 53(53/55), 2(2/55), 0 on the 28 th day in the EA group, and 27(27/55), 25(25/55), 3(3/55) on the 14 th day and 43(43/55), 11(11/55), 1(1/55) on the 28th day in the control group, respectively, with a very significant difference between the two groups (P < 0.01); the EA group in residual urine volume, bladder volume, mean urinary flowing rate was better than the control group on the 14 th day after operation (P < 0.01 or P < 0.05); the hospitalization days after operation was (21.1 +/- 3.3) days in the EA group and (25.5 +/- 3.5) days in the control group, the former being shorter than the later (P < 0.01).

CONCLUSIONEA can promote recovery of bladder function, shorten the keeping time of urinary tube after radical hysterectomy, which is benefit to decreasing incidence rate of urinary system infection and shortening hospitalization days.

Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Hysterectomy ; adverse effects ; Medicine, Chinese Traditional ; Middle Aged ; Postoperative Complications ; physiopathology ; therapy ; Urinary Bladder ; physiopathology ; Urinary Bladder Diseases ; physiopathology ; therapy

OBJECTIVETo observe the acupuncture therapy effect on the urinary retention after radical: hysterectomy. METHODS METHODS: Eighty cases of urinary retention after radical hysterectomy were randomly assigned to: the treatment group and control group according to the random number table method, 40 cases in each group. From the 15th day post operation, the patients in the two groups started to be treated and 7 days as a course with 5 days treatment and 2 days interval. The treatment group applied acupuncture with modalities of common needling on Zusanli (ST36) bilaterally, electroacupuncture on Sanyinjiao (SP6), Shuidao (ST28), and Scalp Reproduction Area bilaterally, moxibustion on Shenque (CV8). The control group applied acupoint injection with vitamin B B12 12, and Sanyinjiao and Zusanli were selected. Take turns on both sides. The courses for the recovery, of bladder function and residual urine volume for those who had voluntary micturition more than 200 mL after the first and second course of treatment were compared between the two groups.

RESULTSWithin 1 course and 2: courses of treatment, the patients with bladder function recovery in the treatment group were 21 (21/40) and 36 (36/40), and those in the control group were 12 (12/40) and 29 (29/40), both with a significant difference (P<0.05). After the first course and second course, residual urine volume for those who had voluntary micturition more than 200 mL in the treatment group was 91.7 ± 17.5 mL and 93.5 ± 15.5 mL, in the control group 102.4 ± 13.7 mL and 102.5 ± 15.7 mL, both with a significant difference (P<0.05).

CONCLUSIONSCombination of acupuncture: modalities was better than acupoint injection for the recovery of bladder function in urinary retention after radical hysterectomy. It would shorten the course of treatment and get a better recovery, decrease urinary retention cases of refractoriness, and extending the treatment could raise the healing rate of urinary retention.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Hysterectomy ; adverse effects ; Middle Aged ; Recovery of Function ; Urinary Bladder ; physiopathology ; Urinary Retention ; etiology ; physiopathology ; therapy ; urine ; Urination ; physiology ; Young Adult